2024/09/20 更新

写真a

カミムラ シンジ
上村 慎治
KAMIMURA Shinji
所属
理工学部 教授
その他担当機関
理工学研究科生命科学専攻博士課程前期課程
理工学研究科生命科学専攻博士課程後期課程
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プロフィール
細胞運動の分子機構、特に、分子モーターや細胞骨格タンパク質の構造と機能がどのように関わっているかに興味を持っています。20億年を越す永い進化の過程で、厳選された理想的な分子構造の基盤が完成しており、その上ではじめて実現される優れた機能があると想像しています。その結果、分子のドメイン構造、その動態はすべて意味を持ったものであると考え、細胞生理学、生物物理学的な独自の手法を駆使して研究を進めています。
外部リンク

学位

  • 理学博士 ( 東京大学 )

  • 理学修士 ( 東京大学 )

学歴

  • 1983年3月
     

    東京大学   理学系研究科   生物科学専攻   博士   修了

  • 1980年3月
     

    東京大学   理学系研究科   修士   修了

  • 1978年3月
     

    東京大学   理学部   生物学科   卒業

経歴

  • 2012年7月 - 2012年9月

    鳥取大学大学院 工学研究科 機械宇宙工学専攻 非常勤講師

  • 2012年7月 - 2012年9月

    鳥取大学大学院 工学研究科 機械宇宙工学専攻 非常勤講師

  • 2005年4月 - 2012年3月

    お茶の水女子大学 大学院理学系研究科 非常勤講師

  • 2005年4月 - 2012年3月

    お茶の水女子大学 大学院理学系研究科 非常勤講師

  • 2011年4月 - 2011年9月

    東京大学理学部 非常勤講師

  • 2006年4月 - 2011年3月

    日本女子大学 理学部 非常勤講師

  • 2008年4月 - 2009年3月

    名古屋大学 工学系研究科 非常勤講師

  • 2008年4月 - 2009年3月

    名古屋大学 工学系研究科 非常勤講師

  • 2008年4月 -  

    中央大学 理工学部 教授

  • 2008年4月 -  

    ~ 中央大学 理工学部 教授

  • 2008年4月 -  

    - Professor at the Department of Biological Sciences Chuo University

  • 2007年4月 - 2008年3月

    東京大学 大学院総合文化研究科 准教授

  • 2007年4月 - 2008年3月

    Associated professor at the Department of Biology, College of Arts and Sciences, University of Tokyo

  • 1998年4月 - 2007年3月

    東京大学 大学院総合文化研究科 助教授

  • 2003年10月 - 2004年3月

    筑波大学生物科学系 非常勤講師

  • 2003年4月 - 2003年9月

    名古屋大学工学系研究科 非常勤講師

  • 2003年4月 - 2003年9月

    名古屋大学工学系研究科 非常勤講師

  • 1992年4月 - 1998年3月

    東京大学 教養学部 助教授

  • 1992年4月 - 1998年3月

    Associated professor at the Department of Biology, College of Arts and Sciences, University of Tokyo

  • 1990年9月 - 1992年3月

    HFSPプログラムフェローシップ(長期派遣)

  • 1990年9月 - 1992年3月

    . HFSP Long-Term Fellowships (LTF)

  • 1985年4月 - 1992年3月

    東京大学 教養学部 助手

  • 1985年4月 - 1992年3月

    Assistant at the Department of Biology, College of Arts and Sciences, University of Tokyo

  • 1985年4月 - 1990年3月

    放送大学 東京学習センター 非常勤講師

  • 1983年4月 - 1985年3月

    新技術開発事業団 水野バイオホロニクスプロジェクト 研究員

▼全件表示

所属学協会

  • (社)日本動物学会

  • 日本生物物理学会

  • Biophysical Society (USA)

  • エアロアクアバイオメカニズム学会

  • 公益社団法人 日本動物学会

  • Biophysical Society (USA)

  • エアロアクアバイオメカニズム学会

  • Zoological Society of Japan

▼全件表示

研究キーワード

  • cilia

  • flagella

  • 鞭毛 繊毛 微小管 細胞運動 細胞骨格 ダイニン シグナル伝達 プライマリ繊毛

  • 構造生物学

  • 細胞生理学

  • 構造生物学

  • ダイニン

  • 細胞骨格

  • 鞭毛

  • 微小管

  • 細胞運動

  • 繊毛

  • プライマリ繊毛

  • Cell physiology

  • シグナル伝達

  • signal transduction

  • primary cilia

  • cytoskeleton

  • dynein

  • microtubules

  • cell motility

研究分野

  • ライフサイエンス / 細胞生物学  / 細胞生物学

  • ライフサイエンス / 細胞生物学  / 細胞生理学

  • ライフサイエンス / 構造生物化学  / 構造生物学

  • ナノテク・材料 / ナノバイオサイエンス

  • ライフサイエンス / 生物物理学  / 生物物理学

  • ナノテク・材料 / ナノ材料科学

▼全件表示

論文

  • Structural insight into the stabilization of microtubules by taxanes

    Andrea E Prota, Daniel Lucena-Agell, Yuntao Ma, Juan Estevez-Gallego, Shuo Li, Katja Bargsten, Fernando Josa-Prado, Karl-Heinz Altmann, Natacha Gaillard, Shinji Kamimura, Tobias Mühlethaler, Federico Gago, Maria A Oliva, Michel O Steinmetz, Wei-Shuo Fang, J Fernando Díaz

    eLIFE   2023年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.7554/eLife.84791

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  • Chemical modulation of microtubule structure through the laulimalide/peloruside site. 査読 国際誌

    Juan Estévez-Gallego, Beatriz Álvarez-Bernad, Benet Pera, Christoph Wullschleger, Olivier Raes, Dirk Menche, Juan Carlos Martínez, Daniel Lucena-Agell, Andrea E Prota, Francesca Bonato, Katja Bargsten, Jelle Cornelus, Juan Francisco Giménez-Abián, Peter Northcote, Michel O Steinmetz, Shinji Kamimura, Karl-Heinz Altmann, Ian Paterson, Federico Gago, Johan Van der Eycken, J Fernando Díaz, María Ángela Oliva

    Structure (London, England : 1993)   31 ( 1 )   88 - 99   2023年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Taxanes are microtubule-stabilizing agents used in the treatment of many solid tumors, but they often involve side effects affecting the peripheral nervous system. It has been proposed that this could be related to structural modifications on the filament upon drug binding. Alternatively, laulimalide and peloruside bind to a different site also inducing stabilization, but they have not been exploited in clinics. Here, we use a combination of the parental natural compounds and derived analogs to unravel the stabilization mechanism through this site. These drugs settle lateral interactions without engaging the M loop, which is part of the key and lock involved in the inter-protofilament contacts. Importantly, these drugs can modulate the angle between protofilaments, producing microtubules of different diameters. Among the compounds studied, we have found some showing low cytotoxicity and able to induce stabilization without compromising microtubule native structure. This opens the window of new applications for microtubule-stabilizing agents beyond cancer treatment.

    DOI: 10.1016/j.str.2022.11.006

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  • Alternative Approaches to Understand Microtubule Cap Morphology and Function 招待

    Oliva, M.A, Gago, F, Kamimura, S, Díaz, J.F

    ACS Omega   2023年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1021/acsomega.2c06926

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  • Metabolomic profiling upon external digestion in larvae of diving beetles: Cybister Curtis, 1827, Dytiscus Linnaeus, 1758, and Hydaticus Leach, 1817 (Coleoptera: Dytiscidae) 査読

    Toshio Inoda, Shinji Kamimura

    Aquatic Insects   2022年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    The SDS.PAGE analysis of prey animals (fly larvae) during external digestion by aquatic predators, the larvae of selected species in dytiscid genera, showed a major component of protein with a molecular weight of ca. 20 kDa. In addition, the analysis of third instar larvae of Cybister brevis Aube, 1838, by nanoLC.ESI.Q-TOF/MS/MS revealed that the digested body fluid includes a polypeptide with a sequence of 97 amino acids corresponding to hemocyanin N (51% matched by Mascot and BLAST searches) and hemocyanin M (12% matched) derived from flies. We also found evidence indicating that beetle larvae repeatedly release digestive enzymes at least twice while consuming the fly bodies. These results suggest that the digested polypeptides were derived from ubiquitous high molecular substances such as arylphorin subunit C223 precursor included in the body fluid of the fly (Calliphora vicina Robineau-Desvoidy, 1830) that were produced during external digestion by diving beetle larvae.

    DOI: 10.1080/01650424.2022.2076883

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  • GTP-dependent formation of straight tubulin oligomers leads to microtubule nucleation 査読 国際誌

    Rie Ayukawa, Seigo Iwata, Hiroshi Imai, Shinji Kamimura, Masahito Hayashi, Kien Xuan Ngo, Itsushi Minoura, Seiichi Uchimura, Tsukasa Makino, Mikako Shirouzu, Hideki Shigematsu, Ken Sekimoto, Benoît Gigant, Etsuko Muto

    Journal of Cell Biology   220 ( 4 )   e202007033   2021年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ROCKEFELLER UNIV PRESS  

    Nucleation of microtubules (MTs) is essential for cellular activities, but its mechanism is unknown because of the difficulty involved in capturing rare stochastic events in the early stage of polymerization. Here, combining rapid flush negative stain electron microscopy (EM) and kinetic analysis, we demonstrate that the formation of straight oligomers of critical size is essential for nucleation. Both GDP and GTP tubulin form single-stranded oligomers with a broad range of curvatures, but upon nucleation, the curvature distribution of GTP oligomers is shifted to produce a minor population of straight oligomers. With tubulin having the Y222F mutation in the β subunit, the proportion of straight oligomers increases and nucleation accelerates. Our results support a model in which GTP binding generates a minor population of straight oligomers compatible with lateral association and further growth to MTs. This study suggests that cellular factors involved in nucleation promote it via stabilization of straight oligomers.

    DOI: 10.1083/jcb.202007033

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  • GTP-Dependent Formation of Straight Oligomers Leads to Nucleation of Microtubules

    Etsuko Muto, Rie Ayukawa, Seigo Iwata, Hiroshi Imai, Shinji Kamimura, Ken Sekimoto, Gigant Benoit

    BIOPHYSICAL JOURNAL   120 ( 3 )   12A - 12A   2021年2月

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    記述言語:英語   出版者・発行元:CELL PRESS  

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  • 剪断流条件下のDNA二重ラセン溶液で観察されたSAXS信号 査読

    藤田 洋介, 箕浦 高子, Longo Alessandro, 和田 祐子, 八木 俊樹, 岩本裕之, 神谷 律, 上村 慎治

    SPring-8/SACLA利用研究成果集   8 ( 3 )   2020年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JASRI  

    SAXS signals from the DNA of salmon spermatozoa in an aqueous solution were analyzed using a rheometer-device, which controlled the rate of shearing during SAXS observations. When shearing rate > 1000 s−1 was applied to the DNA suspension, DNA strands were not aligned as observed with other biological filaments, but a stable diffraction peak approximately ranging 1.7–1.8 [nm−1] of Q-values were observed. We compared our observation with simulated SAXS signals.

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  • 走査型レーザー照明を用いた位相差および暗視野照明顕微鏡法の開発 査読

    佐藤毅, 白根智崇, 和田祐子, 今井洋, 上村慎治

    中央大学理工学研究所論文集   2019 ( 25 )   93 - 103   2020年3月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:中央大学  

    本研究では、光学的には観察試料とは無限の距離に配置された開口絞り面上、対物レンズ後焦点面上でレーザー照明光を高速で円状に走査するコノスコープ面走査型光学系を設計し、スペックルノイズを軽減させる方法を試みたので、ここに報告する。観察試料に珪藻標準試料を用い、位相差像、暗視野像、ともにスペックルノイズを軽減できた。高い観察輝度のため、微細な構造物を観察する目的で用いる暗視野照明法への応用が期待される。

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  • Structural model for differential cap maturation at growing microtubule ends 査読 国際誌

    Juan Estevez-Gallego, Fernando Josa-Prado, Siou Ku, Ruben M. Buey, Francisco A. Balaguer, Andrea E. Prota, Daniel Lucena-Agell, Christina Kamma-Lorger, Toshiki Yagi, Hiroyuki Iwamoto, Laurence Duchesne, Isabel Barasoain, Michel O. Steinmetz, Denis Chretien, Shinji Kamimura, J. Fernando Diaz, Maria A. Oliva

    ELIFE   9   e50155   2020年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELIFE SCIENCES PUBLICATIONS LTD  

    Microtubules (MTs) are hollow cylinders made of tubulin, a GTPase responsible for essential functions during cell growth and division, and thus, key target for anti-tumor drugs. In MTs, GTP hydrolysis triggers structural changes in the lattice, which are responsible for interaction with regulatory factors. The stabilizing GTP-cap is a hallmark of MTs and the mechanism of the chemical-structural link between the GTP hydrolysis site and the MT lattice is a matter of debate. We have analyzed the structure of tubulin and MTs assembled in the presence of fluoride salts that mimic the GTP-bound and GDP•Pi transition states. Our results challenge current models because tubulin does not change axial length upon GTP hydrolysis. Moreover, analysis of the structure of MTs assembled in the presence of several nucleotide analogues and of taxol allows us to propose that previously described lattice expansion could be a post-hydrolysis stage involved in Pi release.

    DOI: 10.7554/eLife.50155

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  • 分子モーター研究の新しい方向 キネシンによる微小管の構造変化

    島 知弘, 森川 真夏, 金城 純一, 神原 丈敏, 上村 慎治, 八木 俊樹, 岩本 裕之, 市村 垂生, 渡邉 朋信, 上村 想太郎, 仁田 亮, 岡田 康志, 廣川 信隆

    日本細胞生物学会大会講演要旨集   69回   39 - 39   2017年5月

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    記述言語:日本語   出版者・発行元:(一社)日本細胞生物学会  

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  • 1P143 タキソールは、急速な微小管内チューブリン周期の伸長を誘導する(11. 分子モーター,ポスター,第52回日本生物物理学会年会(2014年度))

    Kamimura Shinji, Kiyohara Megumi, Nakazawa Nobumasa, Fujita Yosuke, Wada Yuuko, Yagi Toshiki, Iwamoto Hiroyuki

    生物物理   54 ( 1 )   S164   2014年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.54.S164_5

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  • 1P197 鞭毛中心構造による軸糸直径調節を通じたダイニンの活性制御機構(12.細胞生物的課題,ポスター,日本生物物理学会年会第51回(2013年度))

    Yagi Toshiki, Fujita Yosuke, Kamimura Shinji, Iwamoto Hiroyuki

    生物物理   53 ( 1 )   S138   2013年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.53.S138_4

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  • 1P213 繊毛の新機能:シグナル伝達増強作用(細胞生物的課題(接着,運動,骨格,伝達,膜),第48回日本生物物理学会年会)

    Takao Daisuke, Kamimura Shinji

    生物物理   50 ( 2 )   S56 - S57   2010年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.50.S56_6

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  • FRAP Analysis Combined With A Single-cell Electroporation Technique In Sea-urchin Spermatozoa

    Daisuke Takao, Shinji Kamimura

    BIOPHYSICAL JOURNAL   96 ( 3 )   520A - 520A   2009年2月

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    記述言語:英語   出版者・発行元:CELL PRESS  

    DOI: 10.1016/j.bpj.2008.12.2680

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  • 3TA4-02 X線繊維回折による微小管構造動態の解析(細胞生物的課題(接着,運動,骨格,伝達,膜),第47回日本生物物理学会年会)

    Kamimura Shinji, Iwamoto Hiroyuki, Miyashiro Daisuke

    生物物理   49   S56   2009年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.49.S56_4

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  • Erratum: Temperature-dependent regulation of reproduction in the diving beetle Dytiscus sharpi (Coieoptera: Dytiscidae)(Zoological Science(November, 2007), 24, (11), (1115-1121))

    Inoda, T., Tajima, F., Taniguchi, H., Saeki, M., Numakura, K., Hasegawa, M., Kamimura, S.

    Zoological Science   25 ( 5 )   560   2008年5月

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    掲載種別:研究論文(学術雑誌)  

    DOI: 10.2108/zsj.25.560

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  • A new microscope optics for laser dark-field illumination applied to high precision two dimensional measurement of specimen displacement 査読 国際誌

    Noda, N., Kamimura, S.

    Review of Scientific Instruments   79 ( 2 )   023704 - 023704   2008年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER INST PHYSICS  

    With conventional light microscopy, precision in the measurement of the displacement of a specimen depends on the signal-to-noise ratio when we measure the light intensity of magnified images. This implies that, for the improvement of precision, getting brighter images and reducing background light noise are both inevitably required. For this purpose, we developed a new optics for laser dark-field illumination. For the microscopy, we used a laser beam and a pair of axicons (conical lenses) to get an optimal condition for dark-field observations. The optics was applied to measuring two dimensional microbead displacements with subnanometer precision. The bandwidth of our detection system overall was 10 kHz. Over most of this bandwidth, the observed noise level was as small as 0.1 nm/radicalHz.

    DOI: 10.1063/1.2839914

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  • Preface

    Naomi Kato, Shinji Kamimura

    Bio-Mechanisms of Swimming and Flying: Fluid Dynamics, Biomimetic Robots, and Sports Science   2008年

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    掲載種別:研究論文(国際会議プロシーディングス)  

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  • 2P-206 ホヤ精子走化性実測データに基づく精子誘引物質感知機構のシミュレーション解析(細胞生物学的課題(2),第46回日本生物物理学会年会)

    Shiba Kogiku, Miyashiro Daisuke, Kamimura Shinji, Yoshida Manabu

    生物物理   48   S106 - S107   2008年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.48.S106_6

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  • 2P-342 高圧ジャンプX線小角散乱法を用いた細胞骨格フィラメントの安定性とヌクレオチドキャップの研究(バイオイメージング(2),第46回日本生物物理学会年会)

    Fujisawa Tetsuro, Matsuo Hiroshi, Iwasa Mitsusada, Erickson Harold P., Kamimura Shinji, Maeda Yuichiro, Popp David

    生物物理   48   S180   2008年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.48.S180_3

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  • 2P166 真核生物ベン毛軸糸内のラセン構造(分子モーター,口頭発表,第45回日本生物物理学会年会)

    上村 慎治, 岩本 裕之

    生物物理   47   S154   2007年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.47.S154_3

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  • A novel flow-alignment technique for the X-ray diffraction structure analysis of sea-urchin sperm flagella

    Shinji Kamimura, Takaaki Sugiyama, Yasunobu Sugimoto, Katsuzo Wakabayashi

    ZOOLOGICAL SCIENCE   23 ( 12 )   1167 - 1167   2006年12月

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    記述言語:英語   出版者・発行元:ZOOLOGICAL SOC JAPAN  

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  • 2P259 X-ray diffraction from dynein motors and microtubules in seaurchin sperm flagellar axonemes(39. Cell motility,Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

    Kamimura Shinji, Iwamoto Hiroyuki, Fujisawa Tetsuro

    生物物理   46 ( 2 )   S360   2006年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.46.S360_3

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  • Analysis of the flagellar motion of Raphidophyceae, Chattonella antiqua

    Shinji Kamimura, Naomi Katoh, Tatsuro Akiba

    ZOOLOGICAL SCIENCE   22 ( 12 )   1446 - 1446   2005年12月

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    記述言語:英語   出版者・発行元:ZOOLOGICAL SOC JAPAN  

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  • Temperature regulates the summer reproductive diapause in diving beetles, Dytiscus sharpi (Coleoptera : Dytiscidae)

    Toshio Inoda, Fumitada Tajima, Hiroshi Taniguchi, Motoyuki Saeki, Kazuki Numakura, Shinji Kamimura

    ZOOLOGICAL SCIENCE   21 ( 12 )   1326 - 1326   2004年12月

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    記述言語:英語   出版者・発行元:ZOOLOGICAL SOC JAPAN  

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  • A new optical system for the determination of pH change in a sea-urchin sperm cell

    Daisuke Takao, Shinji Kamimura

    ZOOLOGICAL SCIENCE   21 ( 12 )   1283 - 1283   2004年12月

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    記述言語:英語   出版者・発行元:ZOOLOGICAL SOC JAPAN  

    Web of Science

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  • 3P200 X線回折によるウニ精子ベン毛軸糸構造の解析(分子モーター)

    上村 慎治, 若山 純一, 田村 巧, 藤澤 哲郎, 岩本 裕之

    生物物理   44   S239   2004年

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    記述言語:日本語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.44.S239_4

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  • Dynein-ADP as a force-generating intermediate revealed by a rapid reactivation of flagellar axoneme

    Tomomi Tani, Shinji Kamimura

    Biophysical Journal   77 ( 3 )   1518 - 1527   1999年9月

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    掲載種別:研究論文(学術雑誌)  

    Fragmented flagellar axonemes of sand dollar spermatozoa were reactivated by rapid photolysis of caged ATP. After a time lag of 10 ms, axonemes treated with protease started sliding disintegration. Axonemes without protease digestion started nanometer-scale high-frequency oscillation after a similar time lag. Force development in the sliding disintegration was measured with a flexible glass needle and its time course was corresponded well to that of the dynein-ADP intermediate production estimated using kinetic rates previously reported. However, with a high concentration (~80 μM) of vanadate, which binds to the dynein-ADP intermediate and forms a stable complex of dynein-ADP-vanadate, the time course of force development in sliding disintegration was not affected at all. In the case of high frequency oscillation, the time lag to start the oscillation, the initial amplitude, and the initial frequency were not affected by vanadate, though the oscillation once started was damped more quickly at higher concentrations of vanadate. These results suggest that during the initial turnover of ATP hydrolysis, force generation of dynein is not blocked by vanadate. A vanadate-insensitive dynein-ADP is postulated as a force-generating intermediate.

    DOI: 10.1016/S0006-3495(99)76999-7

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  • 微小管の形状ゆらぎとチューブリン分子ゆらぎ 招待

    上村慎治

    細胞   55 ( 3 )   54 - 57   1900年3月

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    担当区分:筆頭著者   記述言語:日本語  

    添付ファイル: THECELL_SKamimura2023Mar.pdf

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書籍等出版物

  • 高等学校理科用 文部科学省検定済教科書 生物 東京書籍(生物 301)

    浅島誠( 範囲: 第3編)

    東京書籍  2024年2月  ( ISBN:4487187494

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    総ページ数:479   記述言語:日本語  

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  • 分子細胞生物学

    Lodish, Harvey F., Berk, Arnold, Kaiser, Chris, Krieger, Monty, Bretscher, Anthony, Ploegh, Hidde L., Martin, Kelsey C., Yaffe, Michael B., Amon, Angelika, 岩井, 佳子, 上村, 慎治, 北川, 大樹, 齋藤, 康太, 坪井, 貴司, 富田, 泰輔, 名黒, 功, 仁科, 博史, 宮澤, 恵二, 山本, 啓一, 若林, 憲一, 堅田, 利明, 須藤, 和夫( 担当: 編訳 範囲: 10.生体膜の構造, 17.細胞の構築と運動 I:ミクロフィラメント, 20.細胞から組織への集成)

    東京化学同人  2023年7月  ( ISBN:9784807920518

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    総ページ数:xxxv, 1074p   記述言語:日本語  

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  • 動物の事典

    上村慎治( 担当: 共編者(共編著者) 範囲: 章担当編集者)

    朝倉書店  2020年11月 

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    総ページ数:746   担当ページ:3ページ   記述言語:日本語   著書種別:事典・辞書

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  • 動物学の百科事典

    上村慎治( 担当: 編集 範囲: 第5章編集、導入、「細胞骨格」担当)

    丸善出版株式会社  2018年9月 

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    総ページ数:770   担当ページ:3   記述言語:日本語  

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  • 動物学の百科事典

    丸善出版株式会社  2018年9月 

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  • 改訂 生物 [2 東書 生物 306] 高校教科書 文部科学省検定済教科書

    浅島, 誠( 範囲: 第3編)

    東京書籍  2018年2月  ( ISBN:4487165555

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    総ページ数:3, 480, 4, 2p   記述言語:日本語  

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  • 改訂 生物基礎 [2東書/生基311] 文部科学省検定済教科書

    浅島, 誠( 範囲: 第3編)

    東京書籍  2017年2月  ( ISBN:4487165490

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    総ページ数:3, 248, 2p   記述言語:日本語  

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  • ケイン生物学

    上村慎治( 担当: 監修 範囲: 動物の反応と調節)

    東京化学同人  2014年9月 

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    総ページ数:728   担当ページ:60   記述言語:日本語   著書種別:学術書

    http://www.tkd-pbl.com/book/b182504.html

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  • Cain Biology

    Tokyo Kagaku Dojin  2014年9月 

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  • ケイン基礎生物学

    上村慎治, 野口朋子, 上村真理子( 担当: 監修 範囲: 動物の反応と調節)

    東京化学同人  2012年2月 

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    総ページ数:360   担当ページ:360   記述言語:日本語   著書種別:学術書

    http://www.tkd-pbl.com/book/b182504.html

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  • Cain Biology

    Tokyo Kagaku Dojin  2012年2月 

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  • 生き物に学ぶ泳ぎと飛行のしくみ(第4章 微生物の運動)

    エアロ・アクアバイオメカニズム研究会(編)  2010年8月 

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  • 生き物に学ぶ泳ぎと飛行のしくみ(第4章 微生物の運動)

    ( 担当: 単著)

    エアロ・アクアバイオメカニズム研究会(編)  2010年8月 

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    記述言語:日本語   著書種別:学術書

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  • 生物学辞典

    石川統, 黒岩常祥, 塩見正衛, 松本忠夫, 守隆夫, 八杉貞雄, 山本正幸( 担当: 共編者(共編著者))

    東京化学同人  2009年10月 

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    総ページ数:1620   担当ページ:40   記述言語:日本語   著書種別:学術書

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  • Bio-mechanisms of swimming and flying: Fluid dynamics, biomimetic robots, and sports science.

    Kato, N, Kamimura,S( 担当: 単著)

    Springer  2007年4月 

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    総ページ数:403   記述言語:英語   著書種別:学術書

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  • Bio-mechanisms of swimming and flying: Fluid dynamics, biomimetic robots, and sports science.

    Springer  2007年4月 

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MISC

  • Kinesin-binding-triggered conformation switching of microtubules contributes to polarized transport. 国際誌

    Tomohiro Shima, Manatsu Morikawa, Junichi Kaneshiro, Taketoshi Kambara, Shinji Kamimura, Toshiki Yagi, Hiroyuki Iwamoto, Sotaro Uemura, Hideki Shigematsu, Mikako Shirouzu, Taro Ichimura, Tomonobu M Watanabe, Ryo Nitta, Yasushi Okada, Nobutaka Hirokawa

    The Journal of cell biology   217 ( 12 )   4164 - 4183   2018年12月

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    記述言語:英語  

    Kinesin-1, the founding member of the kinesin superfamily of proteins, is known to use only a subset of microtubules for transport in living cells. This biased use of microtubules is proposed as the guidance cue for polarized transport in neurons, but the underlying mechanisms are still poorly understood. Here, we report that kinesin-1 binding changes the microtubule lattice and promotes further kinesin-1 binding. This high-affinity state requires the binding of kinesin-1 in the nucleotide-free state. Microtubules return to the initial low-affinity state by washing out the binding kinesin-1 or by the binding of non-hydrolyzable ATP analogue AMPPNP to kinesin-1. X-ray fiber diffraction, fluorescence speckle microscopy, and second-harmonic generation microscopy, as well as cryo-EM, collectively demonstrated that the binding of nucleotide-free kinesin-1 to GDP microtubules changes the conformation of the GDP microtubule to a conformation resembling the GTP microtubule.

    DOI: 10.1083/jcb.201711178

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  • Kinesin-binding-triggered conformation switching of microtubules contributes to polarized transport 国際誌

    Tomohiro Shima, Manatsu Morikawa, Junichi Kaneshiro, Taketoshi Kambara, Shinji Kamimura, Toshiki Yagi, Hiroyuki Iwamoto, Sotaro Uemura, Hideki Shigematsu, Mikako Shirouzu, Taro Ichimura, Tomonobu M. Watanabe, Ryo Nitta, Yasushi Okada, Nobutaka Hirokawa

    JOURNAL OF CELL BIOLOGY   217 ( 12 )   4164 - 4183   2018年12月

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    記述言語:英語   出版者・発行元:ROCKEFELLER UNIV PRESS  

    Kinesin-1, the founding member of the kinesin superfamily of proteins, is known to use only a subset of microtubules for transport in living cells. This biased use of microtubules is proposed as the guidance cue for polarized transport in neurons, but the underlying mechanisms are still poorly understood. Here, we report that kinesin-1 binding changes the microtubule lattice and promotes further kinesin-1 binding. This high-affinity state requires the binding of kinesin-1 in the nucleotide-free state. Microtubules return to the initial low-affinity state by washing out the binding kinesin-1 or by the binding of non-hydrolyzable ATP analogue AMPPNP to kinesin-1. X-ray fiber diffraction, fluorescence speckle microscopy, and second-harmonic generation microscopy, as well as cryo-EM, collectively demonstrated that the binding of nucleotide-free kinesin-1 to GDP microtubules changes the conformation of the GDP microtubule to a conformation resembling the GTP microtubule.

    DOI: 10.1083/jcb.201711178

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  • Simulation of intra-ciliary diffusion suggests a novel role of primary cilia as a cell-signaling enhancer. 査読

    Daisuke Takao, Shinji Kamimura

    Development, growth & differentiation   59 ( 5 )   415 - 422   2017年6月

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    記述言語:英語   出版者・発行元:Develop. Growth Differ.  

    Besides the role to generate a fluid flow in the surrounding medium, eukaryotic cilia have a crucial function in sensing external signals such as chemical or mechanical stimuli. A large body of work has shown that cilia are frequently found in various types of sensory cells and are closely related to many regulatory mechanisms in differentiation and development. However, we do not yet have a definitive answer to the fundamental question, 'why cilia?.' It has been a long-standing mystery why cells use cilia for sensing external signals. To shed light on this, we sought to describe the kinetics of signaling with theoretical approaches. Based on the results, here we propose a new role of cilia as a cell-signaling enhancer. The enhancing effect comes from restricted volume for the free intra-ciliary diffusion of molecules due to the cylindrical shape of cilia, which can facilitate quick accumulation of intracellular signaling molecules. Our simulations demonstrate that both the rate and amplitude of response in signal transduction depend on where the membrane receptors or channels are located along the ciliary shaft. In addition, the calculated transfer function of cilia regarded as a

    DOI: 10.1111/dgd.12360

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  • Simulation of intra-ciliary diffusion suggests a novel role of primary cilia as a cell-signaling enhancer

    Daisuke Takao, Shinji Kamimura

    DEVELOPMENT GROWTH & DIFFERENTIATION   59 ( 5 )   415 - 422   2017年6月

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    記述言語:英語   出版者・発行元:WILEY  

    Besides the role to generate a fluid flow in the surrounding medium, eukaryotic cilia have a crucial function in sensing external signals such as chemical or mechanical stimuli. A large body of work has shown that cilia are frequently found in various types of sensory cells and are closely related to many regulatory mechanisms in differentiation and development. However, we do not yet have a definitive answer to the fundamental question, "why cilia?" It has been a long-standing mystery why cells use cilia for sensing external signals. To shed light on this, we sought to describe the kinetics of signaling with theoretical approaches. Based on the results, here we propose a new role of cilia as a cell-signaling enhancer. The enhancing effect comes from restricted volume for the free intra-ciliary diffusion of molecules due to the cylindrical shape of cilia, which can facilitate quick accumulation of intracellular signaling molecules. Our simulations demonstrate that both the rate and amplitude of response in signal transduction depend on where the membrane receptors or channels are located along the ciliary shaft. In addition, the calculated transfer function of cilia regarded as a transmitter of external signals also suggests the properties of cilia as a signal enhancer. Since such unique composition of receptors and channels in cilia is found in various types of eukaryotic cells, signal enhancing is presumably one of the most essential and conserved roles of cilia.

    DOI: 10.1111/dgd.12360

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  • X-ray fiber diffraction: a tool for understanding the structural dynamics of tubulin dimers in native microtubules.

    上村慎治

    SPring-8/SACLA Research Frontiers 2016   2016   32 - 33   2017年6月

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    記述言語:英語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:JASRI  

    When we are going to know structural details of biological molecules, there would be two possibilities; electron microscopy and X-ray crystallography. Owing to remarkable improvements in cryo-electron microscopy techniques using direct electron detectors, impressive progresses showing high-resolution (0.3 nm) images of various biological molecules in action are now appearing regularly in leading journals. Since details of protein architectures at the atomic scale have been obtained by X-ray crystallography, we can now combine them with electron micrographs to gain more direct insights into the functions of biomolecules than we expected by conventional tools used over a decade ago. This would also be the case for X-ray fiber diffraction/X-ray solution scattering analysis, which we expect will become the third major method used for structural biology.

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  • X-ray fiber diffraction: a tool for understanding the structural dynamics of tubulin dimers in native microtubules.

    Shinji Kamimura

    SPring-8/SACLA Research Frontiers 2016   32 - 33   2017年6月

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  • X-ray fiber diffraction analysis shows dynamic changes in axial tubulin repeats in native microtubules depending on paclitaxel content, temperature and GTP-hydrolysis. 国際誌

    Shinji Kamimura, Yosuke Fujita, Yuuko Wada, Toshiki Yagi, Hiroyuki Iwamoto

    Cytoskeleton (Hoboken, N.J.)   73 ( 3 )   131 - 44   2016年3月

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    記述言語:英語   出版者・発行元:WILEY-BLACKWELL  

    Microtubules are key components of the cytoskeleton in eukaryotic cells. The dynamics between assembled microtubules and free tubulin dimers in the cytoplasm is closely related to the active shape changes of microtubule networks. One of the most fundamental questions is the association of microtubule dynamics with the molecular conformation of tubulin within microtubules. To address this issue, we applied a new technique for the rapid shear-flow alignment of biological filaments, enabling us to acquire the structural periodicity data of microtubules by X-ray fiber diffraction under various physiological conditions. We classified microtubules into three main groups on the basis of distinct axial tubulin periodicities and mean microtubule diameters that varied depending on GTP hydrolysis and the content of paclitaxel, a microtubule stabilizer. Paclitaxel induced rapid changes in tubulin axial repeats in a cooperative manner. This is the first demonstration of dynamic changes of axial tubulin repeats within native microtubules without fixation. We also found extraordinary features of negative thermal expansion of axial tubulin repeats in both paclitaxel-stabilized and GMPCPP-containing microtubules. Our results suggest that even in assembled microtubules, both GTP- and GDP-tubulin dimers can undergo dynamic conversion between at least two different states: short and long configurations. (c) 2016 Wiley Periodicals, Inc.

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  • X-ray fiber diffraction analysis shows dynamic changes in axial tubulin repeats in native microtubules depending on paclitaxel content, temperature and GTP-hydrolysis

    Shinji Kamimura

    Cytoskeleton   73 ( 3 )   131 - 144   2016年3月

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    記述言語:英語   出版者・発行元:WILEY  

    Microtubules are key components of the cytoskeleton in eukaryotic cells. The dynamics between assembled microtubules and free tubulin dimers in the cytoplasm is closely related to the active shape changes of microtubule networks. One of the most fundamental questions is the association of microtubule dynamics with the molecular conformation of tubulin within microtubules. To address this issue, we applied a new technique for the rapid shear-flow alignment of biological filaments, enabling us to acquire the structural periodicity data of microtubules by X-ray fiber diffraction under various physiological conditions. We classified microtubules into three main groups on the basis of distinct axial tubulin periodicities and mean microtubule diameters that varied depending on GTP hydrolysis and the content of paclitaxel, a microtubule stabilizer. Paclitaxel induced rapid changes in tubulin axial repeats in a cooperative manner. This is the first demonstration of dynamic changes of axial tubulin repeats within native microtubules without fixation. We also found extraordinary features of negative thermal expansion of axial tubulin repeats in both paclitaxel-stabilized and GMPCPP-containing microtubules. Our results suggest that even in assembled microtubules, both GTP- and GDP-tubulin dimers can undergo dynamic conversion between at least two different states: short and long configurations. (c) 2016 Wiley Periodicals, Inc.

    DOI: 10.1002/cm.21283

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  • A simple method for nanobubble generation and stability of the bubbles

    Ohmori, M, Haruta, K, Kamimura, S, Koike, H, Uchida, Takeyama, H

    Journal of Environmental Biotechnology   41 - 44   2015年10月

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  • A simple method for nanobubble generation and stability of the bubbles 査読

    Ohmori, M, Haruta, K, Kamimura, S, Koike, H, Uchida, Takeyama, H

    Journal of Environmental Biotechnology   15 ( 1 )   41 - 44   2015年10月

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    記述言語:英語   出版者・発行元:環境バイオテクノロジー学会  

    We could generate very small air-nanobubbles, having diameters smaller than 100 nm, using a household hand mixer. The bubbles were stable in water containing 1% ethanol more than one month and show zeta potential of -30 to -40 mV. The addition of NaCl to nonobubble-water caused changes in the diameter and the number of bubbles, larger in size and less in numbers.

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  • X-Ray Fiber Diffraction Recordings from Oriented Demembranated Chlamydomonas Flagellar Axonemes. 国際誌

    Shiori Toba, Hiroyuki Iwamoto, Shinji Kamimura, Kazuhiro Oiwa

    Biophysical journal   108 ( 12 )   2843 - 53   2015年6月

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    記述言語:英語   出版者・発行元:CELL PRESS  

    The high homology of its axonemal components with humans and a large repertoire of axonemal mutants make Chlamydomonas a useful model system for experiments on the structure and function of eukaryotic cilia and flagella. Using this organism, we explored the spatial arrangement of axonemal components under physiological conditions by small-angle x-ray fiber diffraction. Axonemes were oriented in physiological solution by continuous shear flow and exposed to intense and stable x rays generated in the synchrotron radiation facility SPring-8, BL45XU. We compared diffraction patterns from axonemes isolated from wild-type and mutant strains lacking the whole outer arm (oda1), radial spoke (pf14), central apparatus (pf18), or the a-chain of the outer arm dynein (oda11). Diffraction of the axonemes showed a series of well-defined meridional/layer-line and equatorial reflections. Diffraction patterns from mutant axonemes exhibited a systematic loss/attenuation of meridional/layer-line reflections, making it possible to determine the origin of various reflections. The 1/24 and 1/12 nm(-1) meridional reflections of oda1 and oda11 were much weaker than those of the wild-type, suggesting that the outer dynein arms are the main contributor to these reflections. The weaker 1/32 and 1/13.7 nm(-1) meridional reflections from pf14 compared with the wild-type suggest that these reflections come mainly from the radial spokes. The limited contribution of the central pair apparatus to the diffraction patterns was confirmed by the similarity between the patterns of the wild-type and pf18. The equatorial reflections were complex, but a comparison with electron micrograph-based models allowed the density of each axonemal component to be estimated. Addition of ATP to rigor-state axonemes also resulted in subtle changes in equatorial intensity profiles, which could report nucleotide-dependent structural changes of the dynein arms. The first detailed description of axonemal reflections presented here serves as a landmark for further x-ray diffraction studies to monitor the action of constituent proteins in functional axonemes.

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  • X-Ray Fiber Diffraction Recordings from Oriented Demembranated Chlamydomonas Flagellar Axonemes

    Toba, S., Iwamoto, H., Kamimura, S., Oiwa, K.

    Biophysical Journal   108 ( 12 )   2843 - 2853   2015年6月

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    記述言語:英語   出版者・発行元:CELL PRESS  

    The high homology of its axonemal components with humans and a large repertoire of axonemal mutants make Chlamydomonas a useful model system for experiments on the structure and function of eukaryotic cilia and flagella. Using this organism, we explored the spatial arrangement of axonemal components under physiological conditions by small-angle x-ray fiber diffraction. Axonemes were oriented in physiological solution by continuous shear flow and exposed to intense and stable x rays generated in the synchrotron radiation facility SPring-8, BL45XU. We compared diffraction patterns from axonemes isolated from wild-type and mutant strains lacking the whole outer arm (oda1), radial spoke (pf14), central apparatus (pf18), or the a-chain of the outer arm dynein (oda11). Diffraction of the axonemes showed a series of well-defined meridional/layer-line and equatorial reflections. Diffraction patterns from mutant axonemes exhibited a systematic loss/attenuation of meridional/layer-line reflections, making it possible to determine the origin of various reflections. The 1/24 and 1/12 nm(-1) meridional reflections of oda1 and oda11 were much weaker than those of the wild-type, suggesting that the outer dynein arms are the main contributor to these reflections. The weaker 1/32 and 1/13.7 nm(-1) meridional reflections from pf14 compared with the wild-type suggest that these reflections come mainly from the radial spokes. The limited contribution of the central pair apparatus to the diffraction patterns was confirmed by the similarity between the patterns of the wild-type and pf18. The equatorial reflections were complex, but a comparison with electron micrograph-based models allowed the density of each axonemal component to be estimated. Addition of ATP to rigor-state axonemes also resulted in subtle changes in equatorial intensity profiles, which could report nucleotide-dependent structural changes of the dynein arms. The first detailed description of axonemal reflections presented here serves as a landmark for further x-ray diffraction studies to monitor the action of constituent proteins in functional axonemes.

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  • Effects of the dynein inhibitor ciliobrevin on the flagellar motility of sea urchin spermatozoa

    Yuuko Wada, Shoji A. Baba, Shinji Kamimura

    CYTOSKELETON   72 ( 4 )   182 - 192   2015年4月

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    記述言語:英語   出版者・発行元:WILEY-BLACKWELL  

    Ciliobrevin has recently been found to be a membrane-permeable inhibitor that is specific to AAA+ molecular motors such as cytoplasmic dyneins. In this study, we investigated how ciliobrevin inhibited the motility of sperm from sea urchins: Hemicentrotus pulcherrimus, Pseudocentrotus depressus, and Anthocidaris crassispina. After application of 100 M of ciliobrevin A to live spermatozoa, swimming speed decreased gradually and flagellar motion stopped almost completely within 5 to 10 min. This inhibition was reversible and the frequency of flagellar beating was reduced in a concentration-dependent manner. Ciliobrevin had similar inhibitory effects on the flagellar beating of demembranated and reactivated sperm and the sliding disintegration of trypsin-treated axonemes. We also analyzed the curvature and shear angle of the beating flagella and found that the proximal region of the sperm flagellum was less sensitive to ciliobrevin compared with more distal regions, where bending motions were blocked completely. Interestingly, the shear angle analysis of flagellar motility showed that ciliobrevin induced highly asymmetric bends in the proximal region of the flagellum. These results suggest that there is heterogeneity in the inhibitory thresholds of dynein motors, which depend on the regions along the flagellar shaft (proximal or distal) and on the sites of doublets in the flagellar cross-section (doublet numbers). We expect that it will be possible to map the functional differences in dynein subtypes along and/or around the cross-sections of flagellar axonemes by analyzing the inhibitory effects of ciliobrevin. (c) 2015 Wiley Periodicals, Inc.

    DOI: 10.1002/cm.21218

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  • Effects of the dynein inhibitor ciliobrevin on the flagellar motility of sea urchin spermatozoa. 国際誌

    Yuuko Wada, Shoji A Baba, Shinji Kamimura

    Cytoskeleton (Hoboken, N.J.)   72 ( 4 )   182 - 92   2015年4月

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    記述言語:英語   出版者・発行元:WILEY-BLACKWELL  

    Ciliobrevin has recently been found to be a membrane-permeable inhibitor that is specific to AAA+ molecular motors such as cytoplasmic dyneins. In this study, we investigated how ciliobrevin inhibited the motility of sperm from sea urchins: Hemicentrotus pulcherrimus, Pseudocentrotus depressus, and Anthocidaris crassispina. After application of 100 M of ciliobrevin A to live spermatozoa, swimming speed decreased gradually and flagellar motion stopped almost completely within 5 to 10 min. This inhibition was reversible and the frequency of flagellar beating was reduced in a concentration-dependent manner. Ciliobrevin had similar inhibitory effects on the flagellar beating of demembranated and reactivated sperm and the sliding disintegration of trypsin-treated axonemes. We also analyzed the curvature and shear angle of the beating flagella and found that the proximal region of the sperm flagellum was less sensitive to ciliobrevin compared with more distal regions, where bending motions were blocked completely. Interestingly, the shear angle analysis of flagellar motility showed that ciliobrevin induced highly asymmetric bends in the proximal region of the flagellum. These results suggest that there is heterogeneity in the inhibitory thresholds of dynein motors, which depend on the regions along the flagellar shaft (proximal or distal) and on the sites of doublets in the flagellar cross-section (doublet numbers). We expect that it will be possible to map the functional differences in dynein subtypes along and/or around the cross-sections of flagellar axonemes by analyzing the inhibitory effects of ciliobrevin. (c) 2015 Wiley Periodicals, Inc.

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  • Chemotactic response with a constant delay-time mechanism in Ciona spermatozoa revealed by a high time resolution analysis of flagellar motility

    Daisuke Miyashiro, Kogiku Shiba, Tahahiro Miyashita, Shoji A. Baba, Manabu Yoshida, Shinji Kamimura

    BIOLOGY OPEN   4 ( 2 )   109 - 118   2015年2月

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    記述言語:英語   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    During their chemotactic swimming toward eggs, sperm cells detect their species-specific chemoattractant and sense concentration gradients by unknown mechanisms. After sensing the attractant, sperm cells commonly demonstrate a series of responses involving different swimming patterns by changing flagellar beats, gradually approaching a swimming path toward the eggs, which is the source of chemoattractants. Shiba et al. observed a rapid increase in intracellular Ca2+ concentrations in Ciona spermatozoa after sensing chemoattractants; however, the biochemical processes occurring inside the sperm cells are unclear. In the present study, we focused on the timing and sensing mechanism of chemical signal detection in Ciona. One of the most crucial problems to be solved is defining the initial epoch of chemotactic responses. We adopted a high rate of video recording (600 Hz) for detailed analysis of sperm motion and a novel method for detecting subtle signs of beat forms and moving paths of sperm heads. From these analyses, we estimated a virtual sensing point of the attractant before initiation of motility responses and found that the time delay from sensing to motility responses was almost constant. To evaluate the efficiency of this constant delay model, we performed computer simulation of chemotactic behaviors of Ciona spermatozoa.

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  • Chemotactic response with a constant delay-time mechanism in Ciona spermatozoa revealed by a high time resolution analysis of flagellar motility. 国際誌

    Daisuke Miyashiro, Kogiku Shiba, Tahahiro Miyashita, Shoji A Baba, Manabu Yoshida, Shinji Kamimura

    Biology open   4 ( 2 )   109 - 18   2015年1月

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    記述言語:英語   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    During their chemotactic swimming toward eggs, sperm cells detect their species-specific chemoattractant and sense concentration gradients by unknown mechanisms. After sensing the attractant, sperm cells commonly demonstrate a series of responses involving different swimming patterns by changing flagellar beats, gradually approaching a swimming path toward the eggs, which is the source of chemoattractants. Shiba et al. observed a rapid increase in intracellular Ca2+ concentrations in Ciona spermatozoa after sensing chemoattractants; however, the biochemical processes occurring inside the sperm cells are unclear. In the present study, we focused on the timing and sensing mechanism of chemical signal detection in Ciona. One of the most crucial problems to be solved is defining the initial epoch of chemotactic responses. We adopted a high rate of video recording (600 Hz) for detailed analysis of sperm motion and a novel method for detecting subtle signs of beat forms and moving paths of sperm heads. From these analyses, we estimated a virtual sensing point of the attractant before initiation of motility responses and found that the time delay from sensing to motility responses was almost constant. To evaluate the efficiency of this constant delay model, we performed computer simulation of chemotactic behaviors of Ciona spermatozoa.

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  • Choice of Prey Body Parts for Effective Feeding by Predaceous Diving Beetle Larvae, Dytiscus sharpi sharpi (Wehncke) (Coleoptera: Dytiscidae)

    Toshio Inoda, Shinji Kamimura

    JOURNAL OF INSECT BEHAVIOR   28 ( 1 )   26 - 36   2015年1月

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    記述言語:英語   出版者・発行元:SPRINGER/PLENUM PUBLISHERS  

    Diving beetle larvae use their mandibles in two ways: capturing prey and sucking their body fluid. Catching and consuming the prey's most nutritious body part leads to the highest feeding efficiency. To test this, Dytiscus sharpi sharpi larvae were given tadpoles (Rana ornativentris) as food and their feeding behaviors were observed. Dytiscus larvae preferred to catch tadpoles by the abdomens rather than by other parts. Tadpoles soon became immobilized and in most cases the beetle larvae started eating abdomens first. Beetle larvae tried to change biting site to tadpole's abdomen when the tadpole was initially caught by the head or tail. More food was absorbed from the abdomen than the head or tail suggesting that the feeding behavior of beetle larva is optimized to obtain nutrition efficiently from the tadpole abdomen.

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  • Choice of Prey Body Parts for Effective Feeding by Predaceous Diving Beetle Larvae, Dytiscus sharpi sharpi (Wehncke) (Coleoptera: Dytiscidae)

    Toshio Inoda, Shinji Kamimura

    JOURNAL OF INSECT BEHAVIOR   28 ( 1 )   26 - 36   2015年1月

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    記述言語:英語   出版者・発行元:SPRINGER/PLENUM PUBLISHERS  

    Diving beetle larvae use their mandibles in two ways: capturing prey and sucking their body fluid. Catching and consuming the prey's most nutritious body part leads to the highest feeding efficiency. To test this, Dytiscus sharpi sharpi larvae were given tadpoles (Rana ornativentris) as food and their feeding behaviors were observed. Dytiscus larvae preferred to catch tadpoles by the abdomens rather than by other parts. Tadpoles soon became immobilized and in most cases the beetle larvae started eating abdomens first. Beetle larvae tried to change biting site to tadpole's abdomen when the tadpole was initially caught by the head or tail. More food was absorbed from the abdomen than the head or tail suggesting that the feeding behavior of beetle larva is optimized to obtain nutrition efficiently from the tadpole abdomen.

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  • 微生物の集団運動の物理:遊泳微生物の魅惑の集団ダンス(翻訳)

    上村慎治

    パリティ   28 ( 10 )   30 - 37   2013年10月

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    記述言語:日本語   出版者・発行元:丸善  

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  • 微生物の集団運動の物理:遊泳微生物の魅惑の集団ダンス(翻訳)

    上村慎治

    パリティ   28 ( 10 )   30 - 37   2013年10月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:丸善  

    腸内細菌から海水棲藻類まで,微生|rn|物には集団で協調して泳ぐものが多い。非線形力学,流体力学の理論が,その不思議な行動のしくみに迫る。

    DOI: 10.1063/PT.3.1715

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  • X-ray diffraction recording from single axonemes of eukaryotic flagella. 国際誌

    Masaya Nishiura, Shiori Toba, Daisuke Takao, Daisuke Miyashiro, Hitoshi Sakakibara, Tatsuhito Matsuo, Shinji Kamimura, Kazuhiro Oiwa, Naoto Yagi, Hiroyuki Iwamoto

    Journal of structural biology   178 ( 3 )   329 - 37   2012年6月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    We report the first X-ray diffraction patterns recorded from single axonemes of eukaryotic flagella with a diameter of only <0.2 mu m, by using the technique of cryomicrodiffraction. A spermatozoon isolated from the testis of a fruit fly, Drosophila melanogaster, either intact or demembranated, was mounted straight in a glass capillary, quickly frozen and its 800-mu m segment was irradiated end-on with intense synchrotron radiation X-ray microbeams (diameter, similar to 2 mu m) at 74 K. Well-defined diffraction patterns were recorded, consisting of a large number of isolated reflection spots, extending up to 1/5 nm(-1). These reflections showed a tendency to peak every 20 degrees, i.e., the patterns had features of an 18-fold rotational symmetry as expected from the 9-fold rotational symmetry of axonemal structure. This means that the axonemes remain untwisted, even after the manual mounting procedure. The diffraction patterns were compared with the results of model calculations based on a published electron micrograph of the Drosophila axoneme. The comparison provided information about the native state of axoneme, including estimates of axonemal diameter, interdoublet spacing, and masses of axonemal components relative to those of microtubules (e.g., radial spokes, dynein arms, and proteins associated with accessory singlet microtubules). When combined with the genetic resource of Drosophila, the technique presented here will serve as a powerful tool for studying the structure-function relationship of eukaryotic flagella in general. (c) 2012 Elsevier Inc. All rights reserved.

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  • X-ray diffraction recording from single axonemes of eukaryotic flagella

    Nishiura, M., Toba, S., Takao, D., Miyashiro, D., Sakakibara, H., Matsuo, T., Kamimura, S., Oiwa, K., Yagi, N., Iwamoto, H.

    Journal of Structural Biology   178 ( 3 )   329 - 337   2012年6月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    We report the first X-ray diffraction patterns recorded from single axonemes of eukaryotic flagella with a diameter of only <0.2 mu m, by using the technique of cryomicrodiffraction. A spermatozoon isolated from the testis of a fruit fly, Drosophila melanogaster, either intact or demembranated, was mounted straight in a glass capillary, quickly frozen and its 800-mu m segment was irradiated end-on with intense synchrotron radiation X-ray microbeams (diameter, similar to 2 mu m) at 74 K. Well-defined diffraction patterns were recorded, consisting of a large number of isolated reflection spots, extending up to 1/5 nm(-1). These reflections showed a tendency to peak every 20 degrees, i.e., the patterns had features of an 18-fold rotational symmetry as expected from the 9-fold rotational symmetry of axonemal structure. This means that the axonemes remain untwisted, even after the manual mounting procedure. The diffraction patterns were compared with the results of model calculations based on a published electron micrograph of the Drosophila axoneme. The comparison provided information about the native state of axoneme, including estimates of axonemal diameter, interdoublet spacing, and masses of axonemal components relative to those of microtubules (e.g., radial spokes, dynein arms, and proteins associated with accessory singlet microtubules). When combined with the genetic resource of Drosophila, the technique presented here will serve as a powerful tool for studying the structure-function relationship of eukaryotic flagella in general. (c) 2012 Elsevier Inc. All rights reserved.

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  • Comparative structural analysis of eukaryotic flagella and cilia from Chlamydomonas, Tetrahymena, and sea urchins. 国際誌

    Gaia Pigino, Aditi Maheshwari, Khanh Huy Bui, Chikako Shingyoji, Shinji Kamimura, Takashi Ishikawa

    Journal of structural biology   178 ( 2 )   199 - 206   2012年5月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Although eukaryotic flagella and cilia all share the basic 9 + 2 microtubule-organization of their internal axonemes, and are capable of generating bending-motion, the waveforms, amplitudes, and velocities of the bending-motions are quite diverse. To explore the structural basis of this functional diversity of flagella and cilia, we here compare the axonemal structure of three different organisms with widely divergent bending-motions by electron cryo-tomography. We reconstruct the 3D structure of the axoneme of Tetrahymena cilia, and compare it with the axoneme of the flagellum of sea urchin sperm, as well as with the axoneme of Chlamydomonas flagella, which we analyzed previously. This comparative structural analysis defines the diversity of molecular architectures in these organisms, and forms the basis for future correlation with their different bending-motions. (C) 2012 Elsevier Inc. All rights reserved.

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  • Comparative structural analysis of eukaryotic flagella and cilia from Chlamydomonas, Tetrahymena, and sea urchins

    Pigino, G., Maheshwari, A., Bui, K.H., Shingyoji, C., Kamimura, S., Ishikawa, T.

    Journal of Structural Biology   178 ( 2 )   199 - 206   2012年5月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Although eukaryotic flagella and cilia all share the basic 9 + 2 microtubule-organization of their internal axonemes, and are capable of generating bending-motion, the waveforms, amplitudes, and velocities of the bending-motions are quite diverse. To explore the structural basis of this functional diversity of flagella and cilia, we here compare the axonemal structure of three different organisms with widely divergent bending-motions by electron cryo-tomography. We reconstruct the 3D structure of the axoneme of Tetrahymena cilia, and compare it with the axoneme of the flagellum of sea urchin sperm, as well as with the axoneme of Chlamydomonas flagella, which we analyzed previously. This comparative structural analysis defines the diversity of molecular architectures in these organisms, and forms the basis for future correlation with their different bending-motions. (C) 2012 Elsevier Inc. All rights reserved.

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  • 微生物のべん毛・繊毛の運動メカニズム

    上村慎治

    バイオメカニズム学会誌   34 ( 3 )   176 - 182   2010年8月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:バイオメカニズム学会  

    地球上で現存する2 つの大きな生物グループ,原核生物と真核生物が進化させた2 種類の遊泳運動機構,バクテリアの回転モーター型べん毛と,真核生物の屈曲運動型べん毛・繊毛について,運動メカニズムを紹介し,その力学的な特性を議論する.運動機構の比較から,バクテリア型のべん毛では回転モーターの機械的な強度が,真核生物のべん毛・繊毛ではエネルギー源となるATP 供給速度が,大きな制約条件になっていると考えられる.これは2 つの運動機構がまったく別々の経緯で進化してきた事実と符合する.

    DOI: 10.3951/sobim.34.176

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    その他リンク: https://jlc.jst.go.jp/DN/JALC/00355928752?from=CiNii

  • 微生物の運動

    上村慎治, 曲山幸生, 後藤知伸

    「生きものに学ぶ 泳ぎと飛行のしくみ」   2010年5月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:エアロ・アクアバイオメカニズム研究会編  

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  • Geometry-specific heterogeneity of the apparent diffusion rate of materials inside sperm cells. 国際誌

    Daisuke Takao, Shinji Kamimura

    Biophysical journal   98 ( 8 )   1582 - 8   2010年4月

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    記述言語:英語   出版者・発行元:CELL PRESS  

    In sea urchin spermatozoa, the energy source powering flagellar motion is provided as ATP produced by mitochondria located at the proximal ends of flagella. However, the bottleneck structure between the sperm head and the flagellar tail seems to restrict the free entry of ATP from mitochondria into the tail region. To test this possibility, we investigated the diffusion properties in sperm cells using fluorescence recovery after photobleaching. We found that the rate of fluorescence recovery in the head region was similar to 10% of that observed in the flagellar tail regions. We also found that, even within the tail region, rates varied depending on location, i.e., rates were slower at the more distal regions. Using computational analysis, the rate heterogeneity was shown to be caused mainly by the geometry of the sperm structure, even if little or no difference in diffusion rates through the neck region was assumed. Therefore, we concluded that materials such as ATP would generally diffuse freely between the heads and the flagella of sperm cells. We believe these findings regarding the diffusion properties inside spermatozoa provide further insights into material transportation and chemical signaling inside eukaryotic cilia and flagella.

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  • Geometry-specific heterogeneity of the apparent diffusion rate of materials inside sperm cells

    Takao, D., Kamimura, S.

    Biophysical Journal   98 ( 8 )   1582 - 1588   2010年4月

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    記述言語:英語   出版者・発行元:CELL PRESS  

    In sea urchin spermatozoa, the energy source powering flagellar motion is provided as ATP produced by mitochondria located at the proximal ends of flagella. However, the bottleneck structure between the sperm head and the flagellar tail seems to restrict the free entry of ATP from mitochondria into the tail region. To test this possibility, we investigated the diffusion properties in sperm cells using fluorescence recovery after photobleaching. We found that the rate of fluorescence recovery in the head region was similar to 10% of that observed in the flagellar tail regions. We also found that, even within the tail region, rates varied depending on location, i.e., rates were slower at the more distal regions. Using computational analysis, the rate heterogeneity was shown to be caused mainly by the geometry of the sperm structure, even if little or no difference in diffusion rates through the neck region was assumed. Therefore, we concluded that materials such as ATP would generally diffuse freely between the heads and the flagella of sperm cells. We believe these findings regarding the diffusion properties inside spermatozoa provide further insights into material transportation and chemical signaling inside eukaryotic cilia and flagella.

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  • Single-cell electroporation of fluorescent probes into sea urchin sperm cells and subsequent FRAP analysis 査読

    Takao, D., Kamimura, S.

    Zoological Science   27 ( 3 )   279 - 284   2010年3月

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    記述言語:英語   出版者・発行元:Zoological Society of Japan  

    In sea urchin spermatozoa, the energy required for flagellar motility depends only on the diffusional supply from proximal mitochondria, and thus the diffusion rate inside flagella is one of the most crucial factors limiting the practical size and design of the motile machinery. To determine the diffusion rates of materials inside sperm cells, FRAP (fluorescence recovery after photobleaching) analysis of incorporated fluorescent probes is one of the most powerful approaches. However, the only practically possible method until now was to use the ester forms of fluorescence, and our choice was limited to those of relatively small molecular masses, such as fluorescein derivatives. In this report, we show that a modified single-cell electroporation technique can be applied as a new microinjection method for sperm cells of the sea urchin. The method was applied to FRAP analysis to determine the rate of intraflagellar diffusion.

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  • Single-cell electroporation of fluorescent probes into sea urchin sperm cells and subsequent FRAP analysis

    Daisuke Takao, Shinji Kamimura

    Zoological Science   27 ( 3 )   279 - 284   2010年3月

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    記述言語:英語   出版者・発行元:ZOOLOGICAL SOC JAPAN  

    In sea urchin spermatozoa, the energy required for flagellar motility depends only on the diffusional supply from proximal mitochondria, and thus the diffusion rate inside flagella is one of the most crucial factors limiting the practical size and design of the motile machinery. To determine the diffusion rates of materials inside sperm cells, FRAP (fluorescence recovery after photobleaching) analysis of incorporated fluorescent probes is one of the most powerful approaches. However, the only practically possible method until now was to use the ester forms of fluorescence, and our choice was limited to those of relatively small molecular masses, such as fluorescein derivatives. In this report, we show that a modified single-cell electroporation technique can be applied as a new microinjection method for sperm cells of the sea urchin. The method was applied to FRAP analysis to determine the rate of intraflagellar diffusion.

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  • Structural Changes of Microtubules During GTP Hydrolysis Revealed by X-Ray Fiber-Diffraction

    Shinji Kamimura, Hiroyuki Iwamoto, Daisuke Miyashiro

    BIOPHYSICAL JOURNAL   98 ( 3 )   361A - 361A   2010年1月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:CELL PRESS  

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  • Quick shear-flow alignment of biological filaments for X-ray fiber diffraction facilitated by methylcellulose

    Sugiyama, T., Miyashiro, D., Takao, D., Iwamoto, H., Sugimoto, Y., Wakabayashi, K., Kamimura, S.

    Biophysical Journal   97 ( 12 )   3132 - 3138   2009年12月

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    記述言語:英語   出版者・発行元:CELL PRESS  

    X-ray fiber diffraction is one of the most useful methods for examining the structural details of live biological filaments under physiological conditions. To investigate biologically active or labile materials, it is crucial to finish fiber alignment within seconds before diffraction analysis. However, the conventional methods, e.g., magnetic field alignment and low-speed centrifugations, are time-consuming and not very useful for such purposes. Here, we introduce a new alignment method using a rheometer with two parallel disks, which was applied to observe fiber diffractions of axonemes, tobacco mosaic tobamovirus, and microtubules. We found that fibers were aligned within 5 s by giving high shearflow (1000-5000 s(-1)) to the medium and that methylcellulose contained in the medium (similar to 1%) was essential to the accomplishment of uniform orientation with a small angular deviation (<5 degrees). The new alignment method enabled us to execute structure analyses of axonemes by small-angle x-ray diffraction. Since this method was also useful for the quick alignment of purified microtubules, as well as tobacco mosaic tobamovirus, we expect that we can apply it to the structural analysis of many other biological filaments.

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  • Quick shear-flow alignment of biological filaments for X-ray fiber diffraction facilitated by methylcellulose. 国際誌

    Takaaki Sugiyama, Daisuke Miyashiro, Daisuke Takao, Hiroyuki Iwamoto, Yasunobu Sugimoto, Katsuzo Wakabayashi, Shinji Kamimura

    Biophysical journal   97 ( 12 )   3132 - 8   2009年12月

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    記述言語:英語   出版者・発行元:CELL PRESS  

    X-ray fiber diffraction is one of the most useful methods for examining the structural details of live biological filaments under physiological conditions. To investigate biologically active or labile materials, it is crucial to finish fiber alignment within seconds before diffraction analysis. However, the conventional methods, e.g., magnetic field alignment and low-speed centrifugations, are time-consuming and not very useful for such purposes. Here, we introduce a new alignment method using a rheometer with two parallel disks, which was applied to observe fiber diffractions of axonemes, tobacco mosaic tobamovirus, and microtubules. We found that fibers were aligned within 5 s by giving high shearflow (1000-5000 s(-1)) to the medium and that methylcellulose contained in the medium (similar to 1%) was essential to the accomplishment of uniform orientation with a small angular deviation (<5 degrees). The new alignment method enabled us to execute structure analyses of axonemes by small-angle x-ray diffraction. Since this method was also useful for the quick alignment of purified microtubules, as well as tobacco mosaic tobamovirus, we expect that we can apply it to the structural analysis of many other biological filaments.

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  • Dietary program for rearing the larvae of a diving beetle, Dytiscus sharpi (Wehncke), in the laboratory (Coleoptera: Dytiscidae)

    Inoda, T., Hasegawa, M., Kamimura, S., Hori, M.

    Coleopterists Bulletin   63 ( 3 )   340 - 350   2009年9月

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    記述言語:英語  

    For the conservation of the diving beetle Dytiscus sharpi (Wehncke) (Coleoptera: Dytiscidae), which is included on the Red Data List of Japan, it is critical to understand its ecological background. In the present study, we focused on its feeding behavior and nutritional needs under laboratory breeding conditions. First, we made a list of the possible candidates of prey in the same habitats where we caught D. sharpi. We found that the tadpoles of Rana ornativentris (Werner) were the major species present from March to April, when the beetle larvae appeared. Second, under our laboratory conditions, we investigated the size preference of beetle larvae preying on R. ornativentris tadpoles. We found a significant positive correlation between the developing stage of the larvae and the preferred prey size, i.e., the first and third instars preferred smaller and larger prey, respectively, but second instars did not show any size preference. The size of full-grown adult beetles was almost the same as that of wild insects found in the field, indicating that R. ornativentris tadpoles provide almost complete nutrition for larval growth. Finally, we investigated how the size and number of R. ornativentris tadpoles were correlated with the developing stage of beetle larvae. We suggest that it is crucial for Dytiscus larvae to have access to tadpoles of the proper size and amounts, depending on their growth stage. © 2009 Coleopterists Society.

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  • DIETARY PROGRAM FOR REARING THE LARVAE OF A DIVING BEETLE, DYTISCUS SHARPI (WEHNCKE), IN THE LABORATORY (COLEOPTERA: DYTISCIDAE)

    Toshio Inoda, Masami Hasegawa, Shinji Kamimura, Michio Hori

    COLEOPTERISTS BULLETIN   63 ( 3 )   340 - 350   2009年9月

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    記述言語:英語   出版者・発行元:COLEOPTERISTS SOC  

    For the conservation of the diving beetle Dytiscus sharpi (Wehncke) (Coleoptera: Dytiscidae), which is included on the Red Data List of Japan, it is critical to understand its ecological background. In the present study, we focused on its feeding behavior and nutritional needs under laboratory breeding conditions. First, we made a list of the possible candidates of prey in the same habitats where we caught D. sharpi. We found that the tadpoles of Rana ornativentris (Werner) were the major species present from March to April, when the beetle larvae appeared. Second, under our laboratory conditions, we investigated the size preference of beetle larvae preying on R. ornativentris tadpoles. We found a significant positive correlation between the developing stage of the larvae and the preferred prey size, i.e., the first and third instars preferred smaller and larger prey, respectively, but second instars did not show any size preference. The size of full-grown adult beetles was almost the same as that of wild insects found in the field, indicating that R. ornativentris tadpoles provide almost complete nutrition for larval growth. Finally, we investigated how the size and number of R. ornativentris tadpoles were correlated with the developing stage of beetle larvae. We suggest that it is crucial for Dytiscus larvae to have access to tadpoles of the proper size and amounts, depending on their growth stage. © 2009 Coleopterists Society.

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  • X-ray fiber diffraction studies on flagellar axonemes.

    Oiwa, K., Kamimura, S., Iwamoto, H.

    Methods in cell biology   91 ( 5 )   89 - 109   2009年

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    記述言語:英語   出版者・発行元:ELSEVIER ACADEMIC PRESS INC  

    Eukaryotic cilia and flagella are highly ordered and precisely assembled cellular organelles. Here, to understand the mechanism of the orderly undulations of cilia and flagella, we shall draw a blueprint of their core structures and supporting scaffolds, that is, axonemes, and we shall describe the dynamic structural changes of components of the organelles. Small-angle X-ray scattering and diffraction are among the principal tools used to study protein polymers. These methods are now well established as indispensable tools that complement electron microscopy, providing information on the structure and dynamics of biological materials at atomic resolution in near-physiological environments. For instance, X-ray diffraction studies of skeletal muscles have contributed greatly to our understanding of the structure and molecular mechanisms of muscles. However, owing to the minute size and low diffracting power of axonemes, few attempts at X-ray diffraction of axonemes have been reported. The advent of third-generation synchrotron radiation facilities now makes these attempts feasible, because we now have stable and intense X-rays that enable us to obtain diffractions from the axonemes. In this chapter, we provide a concise practical guide to this new avenue for structural analysis of axonemes.

    DOI: 10.1016/S0091-679X(08)91005-0

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  • X-ray fiber diffraction studies on flagellar axonemes. 国際誌

    Kazuhiro Oiwa, Shinji Kamimura, Hiroyuki Iwamoto

    Methods in cell biology   91   89 - 109   2009年

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    記述言語:英語   出版者・発行元:ELSEVIER ACADEMIC PRESS INC  

    Eukaryotic cilia and flagella are highly ordered and precisely assembled cellular organelles. Here, to understand the mechanism of the orderly undulations of cilia and flagella, we shall draw a blueprint of their core structures and supporting scaffolds, that is, axonemes, and we shall describe the dynamic structural changes of components of the organelles. Small-angle X-ray scattering and diffraction are among the principal tools used to study protein polymers. These methods are now well established as indispensable tools that complement electron microscopy, providing information on the structure and dynamics of biological materials at atomic resolution in near-physiological environments. For instance, X-ray diffraction studies of skeletal muscles have contributed greatly to our understanding of the structure and molecular mechanisms of muscles. However, owing to the minute size and low diffracting power of axonemes, few attempts at X-ray diffraction of axonemes have been reported. The advent of third-generation synchrotron radiation facilities now makes these attempts feasible, because we now have stable and intense X-rays that enable us to obtain diffractions from the axonemes. In this chapter, we provide a concise practical guide to this new avenue for structural analysis of axonemes.

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  • FRAP analysis of molecular diffusion inside sea-urchin spermatozoa

    Daisuke Takao, Shinji Kamimura

    Journal of Experimental Biology   211 ( 22 )   3594 - 3600   2008年11月

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    記述言語:英語   出版者・発行元:COMPANY BIOLOGISTS LTD  

    In sea-urchin spermatozoa, energy required for flagellar motility is provided by ATP diffusion from mitochondria located at the proximal ends of the flagella along with the creatine shuttle system. However, no direct analysis of the diffusion rates inside flagella has been carried out thus far. Using a FRAP (fluorescence recovery after photobleaching) technique, we determined the diffusion coefficients of fluorescein-derivatives (calcein, carboxyfluorescein and Oregon Green) to be 63-64 mu m(2) s(-1). Although these values are about one third of the estimates that were previously used for theoretical calculations, we concluded that the rate of ATP diffusion inside spermatozoa was high enough to support the continuous motility of sea-urchin sperm flagella if the creatine shuttle system is working. We also investigated the diffusion rate through the 'neck' region between the head and tail. When the head region of a calcein-loaded spermatozoon was photobleached, slow recovery of head fluorescence along with the decrease of fluorescence signal in the tail region was observed. It suggests that small molecules such as calcein (M(r), 622.54) can move beyond the boundary between the head and the flagellum. We expect that these findings regarding the diffusion properties inside spermatozoa will provide us with more general insights into the energy equilibrium and material transportation by passive diffusion inside eukaryotic cilia and flagella.

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  • FRAP analysis of molecular diffusion inside sea-urchin spermatozoa 国際誌

    Takao, D., Kamimura, S.

    Journal of Experimental Biology   211 ( 22 )   3594 - 600   2008年11月

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    記述言語:英語  

    In sea-urchin spermatozoa, energy required for flagellar motility is provided by ATP diffusion from mitochondria located at the proximal ends of the flagella along with the creatine shuttle system. However, no direct analysis of the diffusion rates inside flagella has been carried out thus far. Using a FRAP (fluorescence recovery after photobleaching) technique, we determined the diffusion coefficients of fluorescein-derivatives (calcein, carboxyfluorescein and Oregon Green) to be 63-64μm2s-1. Although these values are about one third of the estimates that were previously used for theoretical calculations, we concluded that the rate of ATP diffusion inside spermatozoa was high enough to support the continuous motility of sea-urchin sperm flagella if the creatine shuttle system is working. We also investigated the diffusion rate through the 'neck' region between the head and tail. When the head region of a calcein-loaded spermatozoon was photobleached, slow recovery of head fluorescence along with the decrease of fluorescence signal in the tail region was observed. It suggests that small molecules such as calcein (M r, 622.54) can move beyond the boundary between the head and the flagellum. We expect that these findings regarding the diffusion properties inside spermatozoa will provide us with more general insights into the energy equilibrium and material transportation by passive diffusion inside eukaryotic cilia and flagella.

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  • Temperature-dependent regulation of reproduction in the Diving Beetle Dytiscus sharpi (Coleoptera: Dytiscidae)

    Inoda, T., Tajima, F., Taniguchi, H., Saeki, M., Numakura, K., Hasegawa, M., Kamimura, S.

    Zoological Science   24 ( 11 )   1115 - 21   2007年11月

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    記述言語:英語   出版者・発行元:ZOOLOGICAL SOC JAPAN  

    The effects of temperature on the mating behavior, gonad development, germ cell maturation, and egg spawning of the predaceous diving beetle Dytiscus sharpi (Coleoptera; Dytiscidae), were investigated. By field observations, we found that mating behavior started in October and occurred more frequently from November to December. Under our laboratory breeding conditions, we observed almost the same seasonal variation in mating behavior. We found that temperatures lower than 20 degrees C were required to trigger mating behavior. We also found the same temperature threshold triggered gonadogenesis as well as spermatogenesis. Furthermore, for females, exposure to lower temperatures (<8 degrees C) during the winter was required for egg maturation and spawning in spring; that is, there was a second threshold for successful female reproduction. We conclude that the termination of summer reproductive diapause of D. sharpi is regulated in a temperature-dependent manner, thus effecting the adaptation of D. sharpi to southern warm habitats.

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  • Temperature-dependent regulation of reproduction in the Diving Beetle Dytiscus sharpi (Coleoptera: Dytiscidae)

    Toshio Inoda, Fumitada Tajima, Hiroshi Taniguchi, Motoyuki Saeki, Kazuki Numakura, Masami Hasegawa, Shinji Kamimura

    Zoological Science   24 ( 11 )   1115 - 1121   2007年11月

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    記述言語:英語   出版者・発行元:ZOOLOGICAL SOC JAPAN  

    The effects of temperature on the mating behavior, gonad development, germ cell maturation, and egg spawning of the predaceous diving beetle Dytiscus sharpi (Coleoptera; Dytiscidae), were investigated. By field observations, we found that mating behavior started in October and occurred more frequently from November to December. Under our laboratory breeding conditions, we observed almost the same seasonal variation in mating behavior. We found that temperatures lower than 20 degrees C were required to trigger mating behavior. We also found the same temperature threshold triggered gonadogenesis as well as spermatogenesis. Furthermore, for females, exposure to lower temperatures (<8 degrees C) during the winter was required for egg maturation and spawning in spring; that is, there was a second threshold for successful female reproduction. We conclude that the termination of summer reproductive diapause of D. sharpi is regulated in a temperature-dependent manner, thus effecting the adaptation of D. sharpi to southern warm habitats.

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  • 動物個体の環境応答と情報処理

    上村慎治

    生命科学   239 - 251   2007年2月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:羊土社  

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  • Analysis of small-angle X-ray diffractions from the flow-aligned axonemes of sea-urchin spermatozoa.

    Shinji Kamimura, Hiroyuki Lwamoto, Tetsuro Fujisawa

    BIOPHYSICAL JOURNAL   500A - 500A   2007年1月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:BIOPHYSICAL SOCIETY  

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  • 空と海の生物に学ぶ ―バイオメカニクスの挑戦―

    上村慎治

    遺伝   61 ( 61 )   30 - 32   2007年1月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:エヌ・ティー・エス  

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  • 基礎生命科学実験

    上村慎治

    ( i-ii )   184 - 186   2007年1月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:東京大学出版会  

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  • Dynamic flow alignment of flagellar axonemes for low-angle X-ray fiber diffraction analysis.

    Kamimura, S, Sugiyama, T, Sugimoto, Y, Walabayashi, K

    Photon Factory Actuivity Report 2006   2005-23 ( Part B )   2006年12月

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  • Dynamic flow alignment of flagellar axonemes for low-angle X-ray fiber diffraction analysis.

    Kamimura, S, Sugiyama, T, Sugimoto, Y, Walabayashi, K

    Photon Factory Actuivity Report 2006   2006年12月

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  • Ciliated cells differentiated from mouse embryonic stem cells. 国際誌

    Yusuke Nishimura, Tatsuo S Hamazaki, Shinji Komazaki, Shinji Kamimura, Hitoshi Okochi, Makoto Asashima

    Stem cells (Dayton, Ohio)   24 ( 5 )   1381 - 8   2006年5月

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    記述言語:英語   出版者・発行元:ALPHAMED PRESS  

    In the present study, we demonstrated that the mouse embryonic stem cells were differentiated into ciliated epithelial cells, with characteristics of normal ciliated cells. These cells expressed ciliary marker proteins, such as beta-tubulin IV and hepatocyte nuclear factor-3/forkhead homolog 4 (HFH-4), and processed microtubules were arranged in the 9 + 2 structure, which is the same specific alignment observed in normal ciliary microtubules. The cilia of these cells were beating at a frequency of 17-20 Hz. The differentiated embryoid bodies (EBs) containing these ciliated cells expressed respiratory marker genes such as thyroid transcription factor-1 and surfactant protein-C. For the induction of ciliated cells, culture of EBs in serum-free medium during the initial 2 days of the attachment was indispensable. When EBs were treated with bone morphogenetic proteins, the expression of HFH-4 was decreased, and the ciliated cells were scarcely differentiated. Previous methods for inducing ciliated cells in vitro from embryonic or adult tissues involved an air-liquid interface. The system used in this study more closely mimics the normal development of ciliated cells; thus, an added advantage of the system is as a tool for studying the differentiation mechanism of normal ciliated epithelial cells.

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  • Ciliated cells differentiated from mouse embryonic stem cells

    Nishimura, Y., Hamazaki, T.S., Komazaki, S., Kamimura, S., Okochi, H., Asashimaa, M.

    Stem Cells   24 ( 5 )   1381 - 1388   2006年5月

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    記述言語:英語   出版者・発行元:WILEY  

    In the present study, we demonstrated that the mouse embryonic stem cells were differentiated into ciliated epithelial cells, with characteristics of normal ciliated cells. These cells expressed ciliary marker proteins, such as beta-tubulin IV and hepatocyte nuclear factor-3/forkhead homolog 4 (HFH-4), and processed microtubules were arranged in the 9 + 2 structure, which is the same specific alignment observed in normal ciliary microtubules. The cilia of these cells were beating at a frequency of 17-20 Hz. The differentiated embryoid bodies (EBs) containing these ciliated cells expressed respiratory marker genes such as thyroid transcription factor-1 and surfactant protein-C. For the induction of ciliated cells, culture of EBs in serum-free medium during the initial 2 days of the attachment was indispensable. When EBs were treated with bone morphogenetic proteins, the expression of HFH-4 was decreased, and the ciliated cells were scarcely differentiated. Previous methods for inducing ciliated cells in vitro from embryonic or adult tissues involved an air-liquid interface. The system used in this study more closely mimics the normal development of ciliated cells; thus, an added advantage of the system is as a tool for studying the differentiation mechanism of normal ciliated epithelial cells.

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  • Increase in intracellular pH induces phosphorylation of axonemal proteins for activation of flagellar motility in starfish sperm 国際誌

    Ayako Nakajima, Masaya Morita, Akihiro Takemura, Shinji Kamimura, Makoto Okuno

    Journal of Experimental Biology   208 ( 23 )   4411 - 4418   2005年12月

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    記述言語:英語   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Increased intracellular pH ([pH](i)) activates dynein in sea urchin and mammalian sperm and induces activation of flagellar motility. It is thought that cAMP-dependent protein phosphorylation is associated with motility activation through increasing [pH](i), but little attention has been given to the cAMP-independent phosphorylation also induced by the [pH](i) increase. The present study demonstrates that the increase in [pH](i) in starfish sperm induces the phosphorylation of axonemal proteins and activation of flagellar motility independently of cAMP. Flagellar motility of intact sperm was activated when the [pH](i) was raised by addition of NH4Cl. Histidine, which is known to activate motility of starfish sperm, also raised the [pH](i) during the motility activation. In addition, motility of demembranated sperm flagella was activated in a pH-dependent manner without cAMP. These results indicate that in starfish sperm it is the increase in [pH](i) that induces activation of flagellar motility. Moreover, phosphorylation of axonemal proteins (of molecular mass 25, 32 and 45 kDa) was observed during the pH-dependent and cAMP-independent motility activation of demembranated sperm. This suggests that the increase in [pH](i) regulates flagellar motility via cAMP-independent phosphorylation of axonemal proteins.

    DOI: 10.1242/jeb.01906

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  • A new ratio-imaging technique to measure intracellular pH of sea-urchin sperm flagella

    Daisuke Takao, Shinji Kamimura

    ZOOLOGICAL SCIENCE   22 ( 12 )   1447 - 1447   2005年12月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ZOOLOGICAL SOC JAPAN  

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  • Increase in intracellular pH induces phosphorylation of axonemal proteins for activation of flagellar motility in starfish sperm 査読

    Nakajima, A., Morita, M., Takemura, A., Kamimura, S., Okuno, M.

    Journal of Experimental Biology   208 ( 23 )   4411 - 4418   2005年11月

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    記述言語:英語   出版者・発行元:The Company of Biologists Limited, Cambridge  

    DOI: 10.1242/jeb.01906

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  • New open aquarium system to breed larvae of water beetles (Coleoptera: Dytiscidae)

    Toshio Inoda, Shinji Kamimura

    Coleopterists Bulletin   58 ( 1 )   37 - 43   2004年3月

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    記述言語:英語   出版者・発行元:COLEOPTERISTS SOC  

    For conservation purposes and to supply rare insects for laboratory use, a system for artificial breeding is crucial. However, in the case of carnivorous freshwater insects such as diving beetles, constant conditions in aquariums are difficult to maintain due to their high rate of food consumption. Furthermore, surface rippling caused by the pumping system for water circulation hinders the respiration of small larvae. We developed a new open aquarium system without water circulation that was successfully applied to the rearing of larvae of diving beetles, Dytiscus sharpi (Wehncke) (Coleoptera: Dytiscidae). In comparison to conventional methods, a high proportion of larvae developed into adult insects. The size of reared adults was almost the same as those of field-collected adults. The new method could be applied to the conservation and breeding of other rare species, such as water beetles and water bugs.

    DOI: 10.1649/591

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  • New open aquarium system to breed larvae of water beetles (Coleoptera: Dytiscidae)

    Inoda, T., Kamimura, S.

    Coleopterists Bulletin   58 ( 1 )   37 - 43   2004年3月

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    記述言語:英語   出版者・発行元:COLEOPTERISTS SOC  

    For conservation purposes and to supply rare insects for laboratory use, a system for artificial breeding is crucial. However, in the case of carnivorous freshwater insects such as diving beetles, constant conditions in aquariums are difficult to maintain due to their high rate of food consumption. Furthermore, surface rippling caused by the pumping system for water circulation hinders the respiration of small larvae. We developed a new open aquarium system without water circulation that was successfully applied to the rearing of larvae of diving beetles, Dytiscus sharpi (Wehncke) (Coleoptera: Dytiscidae). In comparison to conventional methods, a high proportion of larvae developed into adult insects. The size of reared adults was almost the same as those of field-collected adults. The new method could be applied to the conservation and breeding of other rare species, such as water beetles and water bugs.

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  • Diameter Oscillation of Axonemes in Sea-Urchin Sperm Flagella

    Sakakibara, H.M., Kunioka, Y., Yamada, T., Kamimura, S.

    Biophysical Journal   86 ( 1 I )   346 - 352   2004年1月

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    記述言語:英語   出版者・発行元:BIOPHYSICAL SOCIETY  

    The 9+2 configuration of axonemes is one of the most conserved structures of eukaryotic organelles. Evidence so far has confirmed that bending of cilia and flagella is the result of active sliding of microtubules induced by dynein arms. If the conformational change of dynein motors, which would be a key step of force generation, is occurring in a three-dimensional manner, we can easily expect that the microtubule sliding should contain some transverse component, i.e., a motion in a direction at a right angle to the longitudinal axis of axonemes. Using a modified technique of atomic force microscopy, we found such transverse motion is actually occurring in an oscillatory manner when the axonemes of sea-urchin sperm flagella were adhered onto glass substrates. The motion was adenosine triphosphate-dependent and the observed frequency of oscillation was similar to that of oscillatory sliding of microtubules that had been shown to reflect the physiological activity of dynein arms (S. Kamimura and R. Kamiya. 1989. Nature. 340: 476-478; 1992. J. Cell Biol. 116: 1443-1454). Maximal amplitude of the diameter oscillation was around 10 nm, which was within a range of morphological change observed with electron microscopy (F.D. Warner. 1978. J. Cell Biol. 77: R19-R26; N. C. Zanetti, D. R. Mitchell, and F. D. Warner. 1979. J. Cell Biol. 80: 573-588).

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  • Diameter Oscillation of Axonemes in Sea-Urchin Sperm Flagella

    Hajime M. Sakakibara, Yuki Kunioka, Takenori Yamada, Shinji Kamimura

    Biophysical Journal   86 ( 1 I )   346 - 352   2004年1月

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    記述言語:英語   出版者・発行元:BIOPHYSICAL SOCIETY  

    The 9+2 configuration of axonemes is one of the most conserved structures of eukaryotic organelles. Evidence so far has confirmed that bending of cilia and flagella is the result of active sliding of microtubules induced by dynein arms. If the conformational change of dynein motors, which would be a key step of force generation, is occurring in a three-dimensional manner, we can easily expect that the microtubule sliding should contain some transverse component, i.e., a motion in a direction at a right angle to the longitudinal axis of axonemes. Using a modified technique of atomic force microscopy, we found such transverse motion is actually occurring in an oscillatory manner when the axonemes of sea-urchin sperm flagella were adhered onto glass substrates. The motion was adenosine triphosphate-dependent and the observed frequency of oscillation was similar to that of oscillatory sliding of microtubules that had been shown to reflect the physiological activity of dynein arms (S. Kamimura and R. Kamiya. 1989. Nature. 340: 476-478; 1992. J. Cell Biol. 116: 1443-1454). Maximal amplitude of the diameter oscillation was around 10 nm, which was within a range of morphological change observed with electron microscopy (F.D. Warner. 1978. J. Cell Biol. 77: R19-R26; N. C. Zanetti, D. R. Mitchell, and F. D. Warner. 1979. J. Cell Biol. 80: 573-588).

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  • Asymmetric Mandibles of Water-Scavenger Larvae Improve Feeding Effectiveness on Right-Handed Snails

    Inoda, T., Hirata, Y., Kamimura, S.

    American Naturalist   162 ( 6 )   811 - 814   2003年12月

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    記述言語:英語   出版者・発行元:UNIV CHICAGO PRESS  

    DOI: 10.1086/378903

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  • Asymmetric Mandibles of Water-Scavenger Larvae Improve Feeding Effectiveness on Right-Handed Snails

    Toshio Inoda, Yoshiyuki Hirata, Shinji Kamimura

    American Naturalist   162 ( 6 )   811 - 814   2003年12月

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    記述言語:英語   出版者・発行元:UNIV CHICAGO PRESS  

    DOI: 10.1086/378903

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  • Effects of Vomeronasal Organ Removal on the Sperm Motility in Male Mice

    Koyama, S., Kamimura, S.

    Zoological Science   20 ( 11 )   1355 - 1358   2003年11月

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    記述言語:英語   出版者・発行元:ZOOLOGICAL SOC JAPAN  

    Odors play important roles in the communication of house mice. They release behaviors and prime changes of the physiological conditions of other individuals. In our previous study, we showed that sperm motility was lowered in the subordinate mice comparing with dominant mice. Our hypothesis is that the lowered sperm motility was due to some primer effects by odor substances derived from dominant mice. To test the hypothesis, we destroyed the vomeronasal organ (VNO) of male mice (VNX male) at 5 weeks of age and paired them with intact male mice (Experimental Group). As control group males, intact male mice were kept in pairs (Control Group). At 15 weeks of age, the sperm motility and weights of reproductive organs, and social dominance was analyzed. The subordinate VNX males were found to have high sperm motility comparable to the dominant males. It was suggested that there is male-to-male primer effects, mediated by VNO, that suppress sperm motility of the subordinate mice.

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  • Study on the development of sperm motility and social dominance of male mice

    Koyama, S., Kamimura, S.

    Physiology and Behavior   80 ( 2-3 )   267 - 272   2003年11月

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    記述言語:英語   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    It has been shown that sperm motility and other parameters related to the reproductive activity varied depending on the social status of male mice. In order to clarify whether such variation is derived from inborn difference or depends on any conditions during maturation, we investigated developmental change of sperm motility, reproductive organs, and body size of MY male mice. We also investigated how the establishment of social dominance of male mice during maturation was correlated with sperm motility. It was found that sperm motility was significantly higher during puberty than in adulthood, although there already existed relatively large statistical variance. The correlation between sperm motility and the social status was revealed to start after 10 weeks of age. It was suggested that a certain inborn difference of sperm motility became enlarged due to environmental factors experienced by male mice during maturation. (C) 2003 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.physbeh.2003.07.001

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  • Study on the development of sperm motility and social dominance of male mice

    Sachiko Koyama, Shinji Kamimura

    Physiology and Behavior   80 ( 2-3 )   267 - 272   2003年11月

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    記述言語:英語   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    It has been shown that sperm motility and other parameters related to the reproductive activity varied depending on the social status of male mice. In order to clarify whether such variation is derived from inborn difference or depends on any conditions during maturation, we investigated developmental change of sperm motility, reproductive organs, and body size of MY male mice. We also investigated how the establishment of social dominance of male mice during maturation was correlated with sperm motility. It was found that sperm motility was significantly higher during puberty than in adulthood, although there already existed relatively large statistical variance. The correlation between sperm motility and the social status was revealed to start after 10 weeks of age. It was suggested that a certain inborn difference of sperm motility became enlarged due to environmental factors experienced by male mice during maturation. (C) 2003 Elsevier Inc. All rights reserved.

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  • Effects of Vomeronasal Organ Removal on the Sperm Motility in Male Mice

    Sachiko Koyama, Shinji Kamimura

    Zoological Science   20 ( 11 )   1355 - 1358   2003年11月

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    記述言語:英語   出版者・発行元:ZOOLOGICAL SOC JAPAN  

    Odors play important roles in the communication of house mice. They release behaviors and prime changes of the physiological conditions of other individuals. In our previous study, we showed that sperm motility was lowered in the subordinate mice comparing with dominant mice. Our hypothesis is that the lowered sperm motility was due to some primer effects by odor substances derived from dominant mice. To test the hypothesis, we destroyed the vomeronasal organ (VNO) of male mice (VNX male) at 5 weeks of age and paired them with intact male mice (Experimental Group). As control group males, intact male mice were kept in pairs (Control Group). At 15 weeks of age, the sperm motility and weights of reproductive organs, and social dominance was analyzed. The subordinate VNX males were found to have high sperm motility comparable to the dominant males. It was suggested that there is male-to-male primer effects, mediated by VNO, that suppress sperm motility of the subordinate mice.

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  • 水薄膜内での精子遊泳観察 査読

    上村慎治

    日本機械学会年次大会講演論文集   ( 6 )   135 - 138   2003年6月

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    記述言語:日本語   出版者・発行元:日本機械学会  

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  • Imaging of the fluorescence spectrum of a single fluorescent molecule by prism-based spectroscopy

    Yoshikazu Suzuki, Tomomi Tani, Kazuo Sutoh, Shinji Kamimura

    FEBS Letters   512 ( 1-3 )   235 - 239   2002年2月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    We have devised a novel method to visualize the fluorescence spectrum of a single fluorescent molecule using prism-based spectroscopy. Equiping a total internal reflection microscope with a newly designed wedge prism, we obtained a spectral image of a single rhodamine red molecule attached to an essential light chain of myosin. We also obtained a spectral image of single-pair fluorescence resonance energy transfer between rhodamine red and Cy5 in a double-labeled myosin motor domain. This method could become a useful tool to investigate the dynamic processes of biomolecules at the single-molecule level. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

    DOI: 10.1016/S0014-5793(02)02269-X

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  • Imaging of the fluorescence spectrum of a single fluorescent molecule by prism-based spectroscopy

    Suzuki, Y., Tani, T., Sutoh, K., Kamimura, S.

    FEBS Letters   512 ( 1-3 )   235 - 239   2002年2月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    We have devised a novel method to visualize the fluorescence spectrum of a single fluorescent molecule using prism-based spectroscopy. Equiping a total internal reflection microscope with a newly designed wedge prism, we obtained a spectral image of a single rhodamine red molecule attached to an essential light chain of myosin. We also obtained a spectral image of single-pair fluorescence resonance energy transfer between rhodamine red and Cy5 in a double-labeled myosin motor domain. This method could become a useful tool to investigate the dynamic processes of biomolecules at the single-molecule level. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

    DOI: 10.1016/S0014-5793(02)02269-X

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  • Effects of social dominance and female odor on sperm activity of male mice

    S Koyama, S Kamimura

    CHEMICAL SIGNALS IN VERTEBRATES 9   9 ( 9 )   403 - 410   2001年

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    記述言語:英語   出版者・発行元:KLUWER ACADEMIC/PLENUM PUBL  

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  • Effects of social dominance and female odor on sperm activity of male mice

    S Koyama, S Kamimura

    CHEMICAL SIGNALS IN VERTEBRATES 9   9   403 - 410   2001年

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  • Influence of social dominance and female odor on the sperm activity of male mice

    Sachiko Koyama, Shinji Kamimura

    Physiology and Behavior   71 ( 3-4 )   415 - 422   2000年11月

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    記述言語:英語   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    In mammals, sperm activity is known to be varied largely according to individuals though physiological reasons have not been clarified yet. In our previous study [Koyama S, Kamimura S. Lowered sperm motility in mice of subordinate social status. Physiol Behav 1999;65:665-669.], we showed that sperm motility was higher in the dominant mice than the subordinate mice, by which it was suggested that social factors could affect sperm activity in mammals. In the present study, we investigated how the observed influence of social dominance would be modified by the existence of females. From 5 to 15 weeks of age, male mice were pair housed and were kept under three different housing conditions: (1) with females; (2) with bedding soiled by females; and (3) control group. The social dominance of the paired males was determined by resident-intruder tests that were carried out from 8 to 15 weeks of age. At the end of 15 weeks of age, sperm activity, weights of organs, level of serum testosterone and corticosterone were determined. It was revealed that sperm density was higher and weight of preputial glands was heavier in dominants than in subordinates when they were kept with females or female bedding, In the subordinates, however, there were no differences among the three housing conditions; that is, there were no female effects on the subordinates. On the other hand, sperm motility was high in the dominants of control group, low in the subordinates, and lower in the dominants that were kept with females. The dominants of the males that were kept with females showed high aggressiveness, and there were negative correlationships to be seen between aggressiveness and sperm motility. It was suggested that: (1) Female odor promotes spermatogenesis of the dominants, but it does not promote that of the subordinates. (2) Sperm motility is more affected by social dominance than by female odor. (3) Excessive aggressiveness has negative influence on sperm motility. (C) 2000 Elsevier Science Inc. All rights reserved.

    DOI: 10.1016/S0031-9384(00)00361-9

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  • Influence of social dominance and female odor on the sperm activity of male mice

    Koyama, S., Kamimura, S.

    Physiology and Behavior   71 ( 3-4 )   415 - 422   2000年11月

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    記述言語:英語   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    In mammals, sperm activity is known to be varied largely according to individuals though physiological reasons have not been clarified yet. In our previous study [Koyama S, Kamimura S. Lowered sperm motility in mice of subordinate social status. Physiol Behav 1999;65:665-669.], we showed that sperm motility was higher in the dominant mice than the subordinate mice, by which it was suggested that social factors could affect sperm activity in mammals. In the present study, we investigated how the observed influence of social dominance would be modified by the existence of females. From 5 to 15 weeks of age, male mice were pair housed and were kept under three different housing conditions: (1) with females; (2) with bedding soiled by females; and (3) control group. The social dominance of the paired males was determined by resident-intruder tests that were carried out from 8 to 15 weeks of age. At the end of 15 weeks of age, sperm activity, weights of organs, level of serum testosterone and corticosterone were determined. It was revealed that sperm density was higher and weight of preputial glands was heavier in dominants than in subordinates when they were kept with females or female bedding, In the subordinates, however, there were no differences among the three housing conditions; that is, there were no female effects on the subordinates. On the other hand, sperm motility was high in the dominants of control group, low in the subordinates, and lower in the dominants that were kept with females. The dominants of the males that were kept with females showed high aggressiveness, and there were negative correlationships to be seen between aggressiveness and sperm motility. It was suggested that: (1) Female odor promotes spermatogenesis of the dominants, but it does not promote that of the subordinates. (2) Sperm motility is more affected by social dominance than by female odor. (3) Excessive aggressiveness has negative influence on sperm motility. (C) 2000 Elsevier Science Inc. All rights reserved.

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  • Dynein-ADP as a force generating intermediate revealed by the rapid reactivation of microtubule sliding in sand-dollar sperm flagella after the photolysis of caged ATP 査読

    Tani, T, Kamimura, S

    Biophys. J.   ( 77 )   1518 - 1527   1999年9月

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    記述言語:英語   出版者・発行元:The Biophysical Society  

    DOI: 10.1016/S0006-3495(99)76999-7

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  • Dynein-ADP as a force generating intermediate revealed by the rapid reactivation of microtubule sliding in sand-dollar sperm flagella after the photolysis of caged ATP

    Tani, T, Kamimura, S

    Biophys. J.   ( 77 )   1518 - 1527   1999年9月

  • 生物光学顕微鏡の基礎

    上村慎治

    生命科学を拓く新しい光技術 (シリーズ・光が拓く生命科学)   28 - 46   1999年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:日本光生物学協会編  

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  • CHEMO-MECHAMCAL COUPLING OF SLIDING BY AXONEMAL DYNEIN(Physiology)(Proceedings of the Seventieth Annual Meeting of the Zoological Society of Japan) :

    Kamimura S., Tani T.

    Zoological science   16   96 - 96   1999年

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    記述言語:英語   出版者・発行元:Zoological Society of Japan  

    CiNii Books

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    その他リンク: http://id.nii.ac.jp/1141/00034671/

  • 軸糸ダイニンATP加水分解反応における滑り方発生中間体

    谷 知己, 上村 慎治

    生物物理   38 ( 2 )   S73   1998年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本生物物理学会  

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  • Reactivation of sea-urchin sperm flagella induced by rapid photolysis of caged ATP

    Tani, T., Kamimura, S.

    Journal of Experimental Biology   201 ( 10 )   1493 - 1503   1998年5月

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    記述言語:英語   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Sea-urchin sperm flagella in a state of rigor were reactivated by rapid photolysis of caged ATP, After a time lag of 11-17 ms, all bends in the axonemes present during rigor began to be propagated towards the tip as if their propagation had not been interrupted. This result suggests that the site-specific activity of dyneins along the length of the axoneme is preserved even during rigor states when ATP is absent and that regulation of the activity can be restarted immediately with a new cycle of ATP turnover. During the starting transient, pre-existing rigor waves in the distal region were propagated without a change in the maximal shear angle until they disappeared at the tip. This was more evident when the rapid reactivation was triggered in high-viscosity solution, in which only the form of new bends was greatly affected by viscous load. After reactivation, the velocity of microtubule sliding increased and reached a plateau within 28 ms. This time course reflects the rate of force generation by dynein in situ.

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  • Reactivation of sea-urchin sperm flagella induced by rapid photolysis of caged ATP

    Tomomi Tani, Shinji Kamimura

    Journal of Experimental Biology   201 ( 10 )   1493 - 1503   1998年5月

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    記述言語:英語   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Sea-urchin sperm flagella in a state of rigor were reactivated by rapid photolysis of caged ATP, After a time lag of 11-17 ms, all bends in the axonemes present during rigor began to be propagated towards the tip as if their propagation had not been interrupted. This result suggests that the site-specific activity of dyneins along the length of the axoneme is preserved even during rigor states when ATP is absent and that regulation of the activity can be restarted immediately with a new cycle of ATP turnover. During the starting transient, pre-existing rigor waves in the distal region were propagated without a change in the maximal shear angle until they disappeared at the tip. This was more evident when the rapid reactivation was triggered in high-viscosity solution, in which only the form of new bends was greatly affected by viscous load. After reactivation, the velocity of microtubule sliding increased and reached a plateau within 28 ms. This time course reflects the rate of force generation by dynein in situ.

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  • 光学顕微鏡入門 - 虫眼鏡から超顕微鏡へ

    上村慎治

    バイオイメージング (シリーズ・ニューバイオフィジックス)   29 - 48   1998年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:日本生物物理学会編  

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  • CHEMO-MECHANICAL COUPLING AND FORCE-GENERATING INTERMEDIATE OF AXONEMAL DYNEIN REVEALED BY A RAPID REACTIVATION EXPERIMENT WITH CAGED-ATP(Physiology)(Proceedings of the Sixty-Ninth Annual Meeting of the Zoological Society of Japan) :

    Tani T., Kamimura S.

    Zoological science   15   100 - 100   1998年

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    記述言語:英語   出版者・発行元:Zoological Society of Japan  

    CiNii Books

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    その他リンク: http://id.nii.ac.jp/1141/00033854/

  • Lowered sperm motility in subordinate social status of mice

    Sachiko Koyama, Shinji Kamimura

    Physiology and Behavior   65 ( 4-5 )   665 - 669   1998年

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    記述言語:英語   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    The correlation between social status and sperm motility of mice was investigated. From 5 to 15 weeks of age, mice were kept under two housing conditions, i.e., in pairs or in isolation. The social dominance in the paired mice was determined with the resident-intruder tests, which were carried out from 8 to 15 weeks of age. At the end of 15 weeks of age, sperm activity, weights of reproductive organs, and serum testosterone were determined. It was revealed that the sperm motility of dominant mice was significantly higher than that of the subordinates. The sperm motility of the isolated mice was also significantly higher than the subordinates. It was suggested that the subordinate social status lowered sperm motility. (C) 1999 Elsevier Science Inc.

    DOI: 10.1016/S0031-9384(98)00205-4

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  • Lowered sperm motility in subordinate social status of mice

    Koyama, S., Kamimura, S.

    Physiology and Behavior   65 ( 4-5 )   665 - 669   1998年

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    記述言語:英語   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    The correlation between social status and sperm motility of mice was investigated. From 5 to 15 weeks of age, mice were kept under two housing conditions, i.e., in pairs or in isolation. The social dominance in the paired mice was determined with the resident-intruder tests, which were carried out from 8 to 15 weeks of age. At the end of 15 weeks of age, sperm activity, weights of reproductive organs, and serum testosterone were determined. It was revealed that the sperm motility of dominant mice was significantly higher than that of the subordinates. The sperm motility of the isolated mice was also significantly higher than the subordinates. It was suggested that the subordinate social status lowered sperm motility. (C) 1999 Elsevier Science Inc.

    DOI: 10.1016/S0031-9384(98)00205-4

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  • 光学顕微鏡で計測する 細胞のミクロ探検 -見えないものを見る-

    上村慎治

    57 - 72   1997年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:日学選書B 日本学術会議事務局・日本動物学会関東支部編集、財団法人 日本学術協力財団発行  

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  • 高分解能顕微鏡で捉えた分子モーターの揺らぎ

    上村慎治

    生体分子モーターのしくみ シリーズ ・ ニューバイオフィジックス   144 - 157   1997年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:日本生物物理学会編  

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  • 光学顕微鏡(1)-光学顕微鏡の基礎と位相差顕微鏡

    上村慎治

    5 ( 2 )   15 - 22   1997年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:遺伝  

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  • 高解像度運動解析

    上村慎治

    細胞   27 ( 8 )   1994年4月

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  • Nanometer scale vibration in mutant axonemes of Chlamydomonas

    Yagi, T., Kamimura, S., Kamiya, R.

    Cell Motility and the Cytoskeleton   29 ( 2 )   177 - 185   1994年

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    記述言語:英語   出版者・発行元:WILEY-LISS  

    Flagellar axonemes of sea urchin sperm display high frequency (200-400 Hz) vibration with nanometer scale amplitudes in the presence of ATP [Kamimura and Kamiya, 1992: J. Cell Biol. 116:1443 -1454]. To investigate how various axonemal components affect the vibration, we examined vibration in wild-type and mutant axonemes of Chlamydomonas. At 1 mM ATP, wild-type axonemes underwent vibration at 100-650 Hz with amplitudes of 4-40 nm. This vibration was similar to, but less regular than, that in sea urchin sperm. Axonemes of the mutants ida1 and ida4 lacking part of the inner arm dynein underwent vibrations indistinguishable from that of wild-type. The mutant oda 1 lacking the entire outer arm underwent vibration at about half the wild-type frequency. Unexpectedly, the paralyzed mutants pf18 lacking the central pair and pf14 lacking the radial spokes displayed vibration with significantly higher frequencies and smaller amplitudes than those in the wild-type vibration. These results indicate that the high-frequency vibration is common to many kinds of mutant axonemes that lack various axonemal substructures, but that its manner is sensitive to the presence of outer arm dynein and the central pair/radial spoke system. Simultaneous measurements of amplitude and frequency in wild-type and mutant axonemes suggest that the velocity of microtubule sliding in vibrating axonemes is lower than the velocity of sliding under load-free conditions. The velocity is particularly low in pf18. A possible mechanism is proposed to explain the lower sliding velocity and vibration amplitude in the pf18 axoneme, based on an assumption that central pair/radial spoke system may work to regulate the switching of two antagonizing forces within the axoneme. (C) 1994 Wiley-Liss, Inc.

    DOI: 10.1002/cm.970290209

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  • Nanometer scale vibration in mutant axonemes of Chlamydomonas

    Toshiki Yagi, Shinji Kamimura, Ritsu Kamiya

    Cell Motility and the Cytoskeleton   29 ( 2 )   177 - 185   1994年

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    記述言語:英語   出版者・発行元:WILEY-LISS  

    Flagellar axonemes of sea urchin sperm display high frequency (200-400 Hz) vibration with nanometer scale amplitudes in the presence of ATP [Kamimura and Kamiya, 1992: J. Cell Biol. 116:1443 -1454]. To investigate how various axonemal components affect the vibration, we examined vibration in wild-type and mutant axonemes of Chlamydomonas. At 1 mM ATP, wild-type axonemes underwent vibration at 100-650 Hz with amplitudes of 4-40 nm. This vibration was similar to, but less regular than, that in sea urchin sperm. Axonemes of the mutants ida1 and ida4 lacking part of the inner arm dynein underwent vibrations indistinguishable from that of wild-type. The mutant oda 1 lacking the entire outer arm underwent vibration at about half the wild-type frequency. Unexpectedly, the paralyzed mutants pf18 lacking the central pair and pf14 lacking the radial spokes displayed vibration with significantly higher frequencies and smaller amplitudes than those in the wild-type vibration. These results indicate that the high-frequency vibration is common to many kinds of mutant axonemes that lack various axonemal substructures, but that its manner is sensitive to the presence of outer arm dynein and the central pair/radial spoke system. Simultaneous measurements of amplitude and frequency in wild-type and mutant axonemes suggest that the velocity of microtubule sliding in vibrating axonemes is lower than the velocity of sliding under load-free conditions. The velocity is particularly low in pf18. A possible mechanism is proposed to explain the lower sliding velocity and vibration amplitude in the pf18 axoneme, based on an assumption that central pair/radial spoke system may work to regulate the switching of two antagonizing forces within the axoneme. (C) 1994 Wiley-Liss, Inc.

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  • 運動をナノメートル精度で測る -顕微計測-

    上村慎治

    実験生物物理(石渡信一編,石川他著)   31 - 44   1993年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:丸善社  

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  • Tubulin protofilaments and kinesin-dependent motility. 査読

    Kamimura, S, Mandelkow, E

    J. Cell Biol.   ( 116 )   1443 - 1454   1992年8月

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    記述言語:英語   出版者・発行元:Rockefeller University Press  

    DOI: 10.1083/jcb.118.4.865

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  • Tubulin protofilaments and kinesin-dependent motility

    S. Kamimura, E. Mandelkow

    Journal of Cell Biology   118 ( 4 )   865 - 875   1992年8月

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    記述言語:英語   出版者・発行元:ROCKEFELLER UNIV PRESS  

    Microtubules are built of tubulin subunits assembled into hollow cylinders which consist of parallel protofilaments. Thus, motor molecules interacting with a microtubule could do so either with one or several tubulin subunits. This makes it difficult to determine the structural requirements for the interaction. One way to approach the problem is to alter the surface lattice. This can be done in several ways. Protofilaments can be exposed on their inside (C-tubules or "sheets"), they can be made antiparallel (zinc sheets), or they can be rolled up (duplex tubules). We have exploited this polymorphism to study how the motor protein kinesin attached to a glass surface interacts and moves the various tubulin assemblies.
    Microtubules glide over the surface along straight paths and with uniform velocities. In the case of C-tubules, approximately 40% glide similarly to microtubules, but a major fraction do not glide at all. This indicates (a) that a full cylindrical closure is not necessary for movement, and (b) that the inside surface of microtubules does not support gliding. With zinc sheets, up to 70% of the polymers move, but the movement is discontinuous, has a reduced speed, and follows along a curved path. Since zinc sheets have protofilaments alternating in orientation and polarity, this result suggests that in principle a single protofilament can produce movement, even when its neighbors cannot. Duplex microtubules do not move because they are covered with protofilaments coiled inside out, thus preventing the interaction with kinesin. The data can be explained by assuming that the outside of one protofilament represents the minimal track for kinesin, but smooth gliding requires several parallel protofilaments. Finally, we followed the motion of kinesin-coated microbeads on sea-urchin sperm flagella, from the flagellar outer doublet microtubules to the singlet microtubule tips extending from the A-tubules. No change in behavior was detected during the transition. This indicates that even if these microtubules differ in surface lattice, this does not affect the motility.

    DOI: 10.1083/jcb.118.4.865

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  • High-frequency vibration in flagellar axonemes with amplitudes reflecting the size of tubulin

    S. Kamimura, R. Kamiya

    Journal of Cell Biology   116 ( 6 )   1443 - 1454   1992年3月

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    記述言語:英語   出版者・発行元:ROCKEFELLER UNIV PRESS  

    Flagellar axonemes of sea urchin sperm display high-frequency (approximately 300 Hz) vibration with nanometer-scale amplitudes in the presence of ATP (Kamimura, S., and R. Kamiya. 1989. Nature (Lond.). 340:476-478). The vibration appears to represent normal mechanochemical interaction between dynein and microtubules because the dependence of the frequency on MgATP concentration is similar to that of the axonemal motility, and because it is inhibited by micromolar concentrations of vanadate. In this study a two-dimensional photo-sensor was used to characterize this phenomenon in detail. Several new features were revealed. First, the vibration was found to be due to a back-and-forth movement of the doublet microtubules along the axonemal length. Two beads attached to different parts of the same axoneme vibrated in unison, i.e., synchronized exactly in phase. This suggested that the outer doublet can be regarded as a stiff rod in vibrating axonemes. Second, evidence was obtained that the amplitude of the vibration reflected the number of active dynein arms. Third, under certain conditions, the vibration amplitude took stepwise values of 8 X N + 4 nm (N = 0, 1, 2, 3, or 4), indicating that the amplitude of microtubule sliding was limited by the size of tubulin dimer (8 nm) or monomer (4 nm). To explain this phenomenon, a model is presented based on an assumption that the force production by dynein is turned off when dynein is subjected to tensile force; i.e., dynein is assumed to be equipped with a feedback mechanism necessary for oscillation.

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  • High-frequency vibration in flagellar axonemes with amplitudes reflecting the size of tubulin

    Kamimura, S., Kamiya, R.

    Journal of Cell Biology   116 ( 6 )   1443 - 1454   1992年

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    記述言語:英語  

    Flagellar axonemes of sea urchin sperm display high-frequency (~300 Hz) vibration with nanometer-scale amplitudes in the presence of ATP (Kamimura, S., and R. Kamiya. 1989. Nature (Lond.). 340: 476-478). The vibration appears to represent normal mechanochemical interaction between dynein and microtubules because the dependence of the frequency on MgATP concentration is similar to that of the axonemal motility, and because it is inhibited by micromolar concentrations of vanadate. In this study a two-dimensional photo- sensor was used to characterize this phenomenon in detail. Several new features were revealed. First, the vibration was found to be due to a back- and-forth movement of the doublet microtubules along the axonemal length. Two beads attached to different parts of the same axoneme vibrated in unison, i.e., synchronized exactly in phase. This suggested that the outer doublet can be regarded as a stiff rod in vibrating axonemes. Second, evidence was obtained that the amplitude of the vibration reflected the number of active dynein arms. Third, under certain conditions, the vibration amplitude took stepwise values of 8 x N + 4 nm (N = 0, 1, 2, 3, or 4), indicating that the amplitude of microtubule sliding was limited by the size of tubulin dimer (8 nm) or monomer (4 nm). To explain this phenomenon, a model is presented based on an assumption that the force production by dynein is turned off when dynein is subjected to tensile force
    i.e., dynein is assumed to be equipped with a feedback mechanism necessary for oscillation.

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  • 微小管

    上村慎治

    バイオメカニクスシリーズ,細胞のバイオメカニクス   143 - 177   1990年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:日本機械学会編,オーム社  

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  • 光学顕微鏡によるnm精度での運動の解析

    上村慎治

    30 ( 4 )   39 - 41   1990年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:生物物理  

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  • High-frequency, nanometre-scale vibration in 'quiescent' flagellar axonemes. 査読

    Kamimura, S, Kamiya, R

    Nature   340 ( 340 )   476 - 478   1989年8月

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    記述言語:英語   出版者・発行元:Nature Publishing Group  

    DOI: 10.1038/340476a0

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  • High-frequency nanometre-scale vibration in 'quiescent' flagellar axonemes

    Shinji Kamimura, Ritsu Kamiya

    Nature   340 ( 6233 )   476 - 478   1989年8月

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    記述言語:英語   出版者・発行元:MACMILLAN MAGAZINES LTD  

    THE movement of cilia and flagella is based on the interaction between dynein arms and microtubules coupled with ATP hydrolysis. Although it is established that dynein arms cause adjacent microtubules to slide, little is known about the elementary process underlying the force production. To look more closely at the mechano-chemical conversion mechanism, we recently developed an optical method for measuring a nanometre-scale displacement with a time-reslution better than 1 ms. We now report the detection of high frequency ( ∼ 300 Hz) vibration of sub-nanometre ampli-tude in non-beating flagellar axonemes. This vibration could reflect the movement of individual activated dynein arms.

    DOI: 10.1038/340476a0

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  • 分子間力を直接測定する新しい方法

    上村慎治

    38 ( 8 )   632 - 633   1989年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:化学と工業  

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  • 光学顕微鏡で細胞運動を高精度で見る

    上村慎治

    4 ( 5 )   56 - 58   1989年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:パリティ  

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  • 細胞運動測定法 nm精度での顆粒運動の解析

    上村慎治

    39 ( 5 )   493 - 495   1988年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:生体の科学  

    DOI: 10.11477/mf.2425905198

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  • Direct measurement of nanometric displacement under an optical microscope

    Shinji Kamimura

    Applied Optics   26 ( 16 )   3425 - 3427   1987年8月

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    記述言語:英語  

    A novel method has been developed to measure nanometric displacement under a conventional optical microscope. The magnified image of a pinhole was divided into two parts using a prism-shaped mirror. The difference of light intensity between the divided images was determined, which was proportional to displacement of the pinhole. Using a 5-Mm diam pinhole, the accuracy to determine displacement was -1 nm. Instead of a pinhole, polystyrene microbeads were used in the new method. Displacement of the microbeads was also measured with nanometric accuracy. This technique could be used to probe nanometric phenomena using optical microscopes. © 1987 Optical Society of America.

    DOI: 10.1364/AO.26.003425

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  • DIRECT MEASUREMENT OF NANOMETRIC DISPLACEMENT UNDER AN OPTICAL MICROSCOPE

    S KAMIMURA

    APPLIED OPTICS   26 ( 16 )   3425 - 3427   1987年8月

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    記述言語:英語   出版者・発行元:OPTICAL SOC AMER  

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  • 新たな細胞運動系、キネシン・チュ-ブリン系の発見

    上村慎治

    生物物理   27 ( 4 )   52 - 54   1987年4月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)   出版者・発行元:生物物理  

    DOI: 10.2142/biophys.27.a180

    CiNii Books

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    その他リンク: https://jlc.jst.go.jp/DN/JALC/00364705832?from=CiNii

  • Turbidimetric studies on microtubule sliding using the stopped-flow-light-scattering method

    Kamimura, S., Nakanishi, M., Yano, M., Shimizu, H.

    Experimental Cell Research   163 ( 1 )   186 - 190   1986年3月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS  

    DOI: 10.1016/0014-4827(86)90571-9

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  • Turbidimetric studies on microtubule sliding using the stopped-flow-light-scattering method

    Shinji Kamimura, Mamoru Nakanishi, Masafumi Yano, Hiroshi Shimizu

    Experimental Cell Research   163 ( 1 )   186 - 190   1986年3月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS  

    The turbidity of axonemes during active sliding of microtubules was analysed using the stopped-flow-light-scattering method with high time resolution. Flagella of sea-urchin spermatozoa were demembranated and used after a brief treatment with trypsin. The turbidity of the suspension of flagellar axonemes during ATP-induced disintegration was measured and its time course fitted to a single exponential function which yielded the rate of disintegration, R(l/sec). R coincided well with the velocity of microtubule sliding, V(μfx186-1 sec) as determined by cinematomicrographic analysis [13], i.e., R = 0.22 × V, r = 0.9973. It indicates that turbidimetry is a useful method with which to learn the sliding velocity of microtubules. From the dependency of R on temperature, Q10 of the sliding velocity was estimated to be 2.0-2.3 at 43-820 μM of MgATP. © 1986.

    DOI: 10.1016/0014-4827(86)90571-9

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  • ATP hydrolysis coupled to microtubule sliding in sea-urchin sperm flagella

    Shinji Kamimura, Masafumi Yano, Hiroshi Shimizu

    Journal of Biochemistry   97 ( 5 )   1509 - 1515   1985年5月

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    記述言語:英語   出版者・発行元:JAPANESE BIOCHEMICAL SOC  

    Using sea urchin (Hemicentrotus pulcherimus) sperm flagella, ATP hydrolysis coupled to sliding movement of microtubules was investigated. Flagellar axonemes were pretreated with trypsin and the microtubules induced to slide by addition of ATP (50-1,000 μM) at 0-20°C. Motion-dependent hydrolysis of ATP was observed immediately after the addition of ATP, the rate of which was higher than that of steady state hydrolysis in axonemes without trypsin-treatment, or after complete disintegration. The rate of hydrolysis of ATP divided by the sliding velocity of microtubules reflects the ATP consumption necessary per unit distance of microtubule sliding. This parameter varied according to the experimental conditions in that it increased when the ATP concentration or temperature was decreased. Our results suggest that there is not a strict stoichiometric relationship between ATP hydrolysis and sliding distance in the dynein-tubulin system, indicating that the mechanochemical coupling is different from that in beating axonemes. © 1985, by the Japanese Biochemical Society.

    DOI: 10.1093/oxfordjournals.jbchem.a135206

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  • ATP hydrolysis coupled to microtubule sliding in sea-urchin sperm flagella

    Kamimura, S., Yano, M., Shimizu, H.

    Journal of Biochemistry   97 ( 5 )   1509 - 1515   1985年

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    記述言語:英語   出版者・発行元:JAPANESE BIOCHEMICAL SOC  

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  • Dynamic aspects of microtubule sliding in sperm flagella

    K. Takahashi, S. Kamimura

    Journal of Submicroscopic Cytology   15 ( 1 )   1 - 3   1983年

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    記述言語:英語   出版者・発行元:EDITRICE COMPOSITORI BOLOGNA  

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  • Dynamic aspects of microtubule sliding in sperm flagella

    Takahashi, K., Kamimura, S.

    Journal of Submicroscopic Cytology   15 ( 1 )   1 - 3   1983年

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    記述言語:英語   出版者・発行元:EDITRICE COMPOSITORI BOLOGNA  

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  • MICROTUBULE SLIDING IN REACTIVATED FLAGELLA

    K TAKAHASHI, C SHINGYOJI, S KAMIMURA

    SYMPOSIA OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY   ( 35 )   159 - 177   1982年

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    記述言語:英語   出版者・発行元:GEORG THIEME VERLAG  

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  • Microtubule sliding in reactivated flagella.

    K. Takahashi, C. Shingyoji, S. Kamimura

    Symposia of the Society for Experimental Biology   35 ( 35 )   159 - 177   1982年

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:GEORG THIEME VERLAG  

    Recent experimental studies of microtubule sliding in demembranated sea urchin sperm flagella are described. A local iontophoretic application of ATP to a Triton-extracted flagellum elicits a local bending response whose form is in exact conformity with the predictions of the sliding microtubule model. Cinematographic analysis of the microtubule sliding initiated by treating fragments of demembranated flagella with trypsin in the presence of ATP reveals that the speed of sliding is almost constant. This implies that the speed does not depend on the number of dynein arms participating in the generation of sliding force. The distribution of apparent sliding velocities indicates that there is no difference in sliding velocity among the doublets. The sliding velocity depends on MgATP concentration in a manner consistent with Michaelis-Menten kinetics. The sliding velocity of doublets in trypsin-treated axonemes is close to the maximum velocity of relative sliding taking place between adjacent doublets in beating flagella reactivated at the same MgATP concentration.

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  • Direct measurement of the force of microtubule sliding in flagella

    Shinji Kamimura, Keiichi Takahashi

    Nature   293 ( 5833 )   566 - 568   1981年

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    記述言語:英語   出版者・発行元:MACMILLAN MAGAZINES LTD  

    The movement of eukaryotic cilia and flagella is caused by ATP-driven active sliding between the doublet microtubules1-3, and the force for the sliding is believed to be generated by the dynein arms of the A-tubule interacting with the B-tubule of the adjoining doublet. To understand the mechanochemical basis of this force-generating reaction and to correlate it with the overt motile behaviour of cilia or flagella, it is important to quantify the force exerted by the dynein arms. Attempts have been made to estimate this force from the bending moment generated by the whole organelle4,5, but the estimation was of necessity hypothetical because the mechanism by which sliding is coupled with bending is poorly understood. Microtubule sliding without bending can be induced in vitro if trypsin-2 or elastase 6-treated axonemes are perfused with a solution containing ATP. By attaching glass microneedles to such axonemes, we have now directly determined the sliding force at various ATP concentrations. © 1981 Nature Publishing Group.

    DOI: 10.1038/293566a0

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  • Direct measurement of the force of microtubule sliding in flagella

    Kamimura, S., Takahashi, K.

    Nature   293 ( 5833 )   566 - 568   1981年

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    記述言語:英語   出版者・発行元:MACMILLAN MAGAZINES LTD  

    DOI: 10.1038/293566a0

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▼全件表示

講演・口頭発表等

  • キイロショウジョウバエの寿命と活動量解析のための新画像処理法

    兵藤 律希, 青木 涼真, 王 舒誠, 加藤 榛名, 上村 慎治

    日本動物学会 第94回 山形大会  2023年9月 

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    開催年月日: 2023年9月    

    記述言語:日本語   会議種別:口頭発表(一般)  

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  • タコノマクラの緑色色素の特性解析

    城山 篤宏, 西田 智弥, 河野 美都子, 加藤 佑亮, 上村 慎治

    日本動物学会 第94回 山形大会  2023年9月 

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    開催年月日: 2023年9月    

    記述言語:日本語   会議種別:口頭発表(一般)  

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  • タコノマクラ緑色色素の特性

    西田智弥, 河野美都子, 加藤佑亮, 上村慎治

    第93回 日本動物学会年次大会 1C23-1630  ( 早稲田大学 )   2022年9月  日本動物学会

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    記述言語:日本語   会議種別:口頭発表(一般)  

    タコノマクラ(Clypeaster japonicus)は、棘皮動物ウニ綱不正形類のタコノマクラ目に属するウニで、物理的な外的損傷や神経刺激、環境の悪化によるストレスなどで、緑色の色素を多量に産生する。特に表皮に局所的な外傷を与えた場合、その部分から緑色色素を滲出させ、傷口を保護するかのような反応が観察されるため、この色素は、広義の生体防御反応に深く関わると我々は考えている。これまでウニから同定された色素は、現在までに40種類以上記載されている。例えば、殻や棘、内臓の色素沈着に関与するスピノクロムまたはエキノクロムと呼ばれるナフトキノン類を産生する種の報告がある。これらの色素の生物学的役割は、抗菌性バイオフィルム形成やUV損傷に対する保護、免疫防御を持つ色素細胞への関与などが示唆されている。医療分野においては高抗酸化能をもつスピノクロムが様々な疾患に対する新たな医薬品の材料として検討され、最近ではエキノクロムAが、ヒストクロムと呼ばれる薬品の活性成分として導入され、眼科疾患および心筋梗塞に対する予防薬として投与されている。このようにウニからは医療分野において応用可能な色素が多く見つかる中、タコノマクラ由来の色素ついての研究報告はきわめて少なく(Goodwin & Fox, 1955)、その成分については明らかになっていない。そのため本研究では色素の持つ特性を調査し、成分分析の解明を目標とした。これまで、吸収スペクトルのpH依存性や長期保存による酸化還元の構造変化、見かけ上の分子量が100 kDa以上であること、加熱処理や酸化によって色素が退色する傾向があること、塩類が発色には直接関係していないこと、緑色色素には有機物構成元素を多く含むこと、同じタコノマクラ目のウニであるスカシカシパン(Astriclypeus manni)との緑色色素に高い類似性があることなどがわかって来た。この色素を分泌する機構も含めて、これまで当研究室で調べられた特性について、紹介する。

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  • タコノマクラ幼生における緑色色素産生細胞

    天野 博人, 河野 美都子, 加藤 佑亮, 上村 慎治

    第93回 日本動物学会年次大会 IC24-1645  ( 早稲田大学 )   2022年9月  日本動物学会

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    記述言語:日本語   会議種別:口頭発表(一般)  

    タコノマクラ(Clypeaster japonicus)は、棘皮動物ウニ綱不正形類のタコノマクラ目に属するウニで、物理的な損傷や環境ストレスなどの刺激によって、緑色の色素(以下、緑色色素と表記する)を産生し体外に分泌するユニークな特性を持つ。表皮に外傷を負った場合、その部分から次第に緑色色素が滲出され、傷口部分に凝集して傷口を覆うような反応が起こることが観察されたため、この色素は、広義の免疫学的な生体防御機構に関わるものではないかと我々は考えている。これまでにウニ類から抽出・同定された色素としてエキノクロムが知られているが(Sadek et al. 2022)、その色素の中でも、卵,卵巣,殻,トゲ,およびそのほかの内臓器官に広く分布している物質(エキノクロムA)がヒストクロムと呼ばれる薬品の活性成分として導入され、実際に眼科疾患および心筋梗塞に対する予防薬として投与されている(Sayed et al. 2018)。それ以外にもウニから医療分野への利用の可能性のある色素が報告されているが、タコノマクラ由来の緑色色素に関しての研究報告はほとんどなく(Goodwin & Fox, 1955)、成分や産生・分泌組織についてはまったく知られていない。この緑色色素を産生する細胞を同定する目的で、本研究を行った。これまで、以下の様な事実が当研究室の研究で明らかになっている(未発表)。成体・幼生共に飼育水中、あるいは、体腔中に注入した0.55M KClによる刺激によって緑色色素の分泌が認められており、8腕プルテウス幼生においては、アセチルコリンによる分泌誘導、アトロピンによる分泌抑制も確認されている。酸性側で水溶性が高く、タンパク質や脂質は含まない物質である。 成体へと発生していく途中のウニ幼生は未分化の細胞が複雑な組織へと分化する過程であるため、体の組織構成が比較的単純であり、緑色色素を産生し分泌する組織や細胞を調べることで成体での色素産生組織を同定するには好都合の実験材料であると考え、本実験では4腕プルテウス幼生~8腕プルテウス幼生を対象としてKClを用いた刺激を行い、その前後の様子を観察することにした。その結果、8腕プルテウス幼生において緑色色素の産生との関与が疑われる色素胞(緑色色素胞と命名)が新たに発見された。この緑色色素胞はタコノマクラ8腕幼生の原基付近に存在し、形にばらつきのある15μm前後の大きさの細胞で、0.55 M KClによる刺激で全て崩壊するかのように消失する特性を持ち、その反応の際には体外へ緑色色素が放出されていることが確認できた。さらに色素胞の形状や構造をくわしく調べる目的で、Acridine orange(AO)等の蛍光染色し、細胞内外の構造を調べたのでここで報告する。

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  • 画像処理を用いたショウジョウバエの寿命解析

    青木涼真, 兵藤律希, 加藤榛名, 王 舒誠, 上村慎治

    第93回 日本動物学会年次大会 3H36-1515  ( 早稲田大学 )   2022年9月  日本動物学会

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    記述言語:日本語   会議種別:口頭発表(一般)  

    キイロショウジョウバエは、飼育が容易なモデル生物である。また、1世代が1~2ヶ月と短いことから、寿命の解析に適した実験材料である。これまで様々な文献で寿命測定の報告があるが、文献ごとに寿命測定の方法にはばらつきがあり、死亡時を特定する方法、誤差、精度も明確ではないことが多い。そのため寿命決定の精度を高くする意義は高いと考えている。さらに各個体の活動量と寿命とは何らかの相関があると予測されるが、既存の寿命解析法では活動量を同時に調べることは難しかった。本研究ではキイロショウジョウバエの寿命をより正確に測定でき、あわせて活動量の変化も測定できる新たな手法を開発することを目的とした。本研究ではキイロショウジョウバエが入ったバイアル瓶の前にカメラを設置し一定間隔で継続的に写真撮影する手法を用いた。また撮影した画像から、正確な寿命が求められるだけでなく、活動量を定量化も試みた。解決すべき課題は、非常に多数(1万枚以上)の撮影画像の処理を手作業ではなく、客観的に再現性良く高速(1枚10秒)に実施することである。我々は撮影した画像からキイロショウジョウバエの画像を自動的に識別し抽出し、個々の位置を計測する自動処理ソフトウェアの開発も同時に行い人口餌に入れたカフェインとアルギニンがキイロショウジョウバエ(Drosophila melanogaster)の寿命に与える影響についての予備的な調査を行った。その結果、他の個体との重複や飛翔中などの理由でうまく撮影できなかった個体がある頻度で出現するが、実用上は問題なく解析が可能であることがわかり、上の2種の添加物の効果の差も明確に議論できることが分かった。照明方法や撮影頻度を最適化すると同時にさらなる画像処理の高速化によってより、キイロショウジョウバエの寿命・行動解析において、有効な手法になると期待している。

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  • ネガティブ染色電子顕微鏡法により明らかにされた繊毛ダイニンの新規構造

    雷 宜慈, 今井 洋, 山本 遼介, 下 理恵子, 上村 慎治, 八木 俊樹, 梶村 直子, 廣瀬 未果, 加藤 貴之, 光岡 薫, 昆 隆英

    第59回 日本生物物理学会年会  ( On Line )   2021年11月  日本生物物理学会

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    記述言語:英語   会議種別:ポスター発表  

    Dyneins are phylogenetically classified into three subfamilies, ciliary, cytoplasmic and intraflagellar transport (IFT) dyneins. It is widely known that cytoplasmic and IFT dyneins adopt an inactivated conformation called the phi structure. In contrast, such a distinct inactivation state was not evident for ciliary dynein. Here we report that one of the ciliary dynein species has a novel structure that is very similar to the phi structure. Based on the structural analysis, we propose a novel shutdown mechanism common to all three subfamilies of dynein.

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  • 低温条件下での微小管構造動態解析:溶液温度に依存した構造変化における非等方性とヒステリシス

    上村慎治, 八木俊樹, 近藤裕祐, Estévez-Gallego J, Lucena-Agell D, Díaz J. F, 岩本 裕之

    第59回 日本生物物理学会年会  ( On Line )   2021年11月  日本生物物理学会

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    記述言語:日本語   会議種別:ポスター発表  

    Microtubules (MT) are involved in essential cellular functions. One known characteristic of them is rapid disassembly to tubulin subunits upon cooling, a process whose molecular basis is not yet understood. We hypothesize that the conformational changes in tubulin accumulate strain of MT that triggers disassembly. To test this hypothesis, we analyzed the X-ray fiber diffraction patterns of native MTs during rapid cooling from 37 to 10℃ (<20s). We found that shape changes of MT on cooling was anisotropic, i.e., the magnitude of shrinkage was different between longitudinal and diameter directions. Detailed time-course analysis showed that the shrinkage and expansion are hysteretic. The present study would help us to understand how MTs become instable at low temperatures.

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  • 赤潮の原因藻類シャトネラの生物対流現象

    中原 美奈, 小林 篤人, 正, 上村 慎治

    第43回エアロ・アクアバイオメカニズム学会講演会  ( ネット開催 )   2021年3月  エアロ・アクアバイオメカニズム学会講演会

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    記述言語:日本語   会議種別:口頭発表(一般)  

    Chattonella is marine algae that causes HAB at Seto Inland Sea in Japan. When collected in a shallow petri dish, they start gradual accumulation in a few minutes and forms specific spotted or branching patterns with nonhomogeneous cell distributions. Since this phenomenon of bioconvection is expected to be tightly correlated with HAB mechanisms, we are executing the image analysis of accumulation behavior of Chattonella using a spatial statistical technique. Examples to show novel quantitative descriptions of bioconvection we found are shown.

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  • Variogram/Correlogram法を使った生物対流解析

    中原美奈, 小林篤人, 上村慎治

    日本生物物理学会  ( Online )   2020年9月  第58回 日本生物物理学会年会

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    記述言語:英語   会議種別:ポスター発表  

    ラフィド藻のChattonellaは、有害な赤潮(HAB)を引き起こす藻類である。実験室内で浅いシャーレに入れて観察すると、短時間で不均一な分布の特定の生物対流パターンを形成する。この現象は,屋外でのHAB形成機構と深く関連していると考えられる。HAB発生のメカニズムを理解するために、空間統計技術と高速ビデオ顕微鏡観察によりChattonella marina var. ovata(Raphidophyceae)の集団的遊泳行動の画像解析を行ったので,ここで紹介する。

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  • X線繊維回折法によって明らかとなった秒単位の微小管構造変化 招待

    上村慎治

    宮崎シーガイアコンベンションセンター  ( 宮崎 )   2019年9月  日本生物物理学会

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    記述言語:英語   会議種別:口頭発表(一般)  

    Microtubules are assembled from tubulin dimers that are arranged in a semi-crystal lattice with high regularity. Dynamics of tubulin dimer within microtubules is thus expected to be directly affecting the stability of microtubules, however, it has been difficult to quantitatively describe such properties. By analyzing the time course of pattern changes in X-ray fiber diffractions from aligned microtubules with a high-flux synchrotron beam of BM40XL (SPring-8), we found microtubules showed rapid elongation in the axial tubulin repeat and the concomitant increase of structural flexibility with a time constant of about 0.2 s after applying paclitaxel, microtubule-stabilizer. Contrasting effects were found for laulimalide, another type of microtubules stabilizer.

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  • 微小管内チューブリン周期の絶対値

    上村慎治

    日本動物学会 第90回 大阪大会  ( 大阪市立大学 )   2019年9月  日本動物学会

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    記述言語:日本語   会議種別:口頭発表(招待・特別)  

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  • 微小管内のチューブリン分子の安定性・可塑性・柔軟性を目安にした微細動態解析

    上村慎治, 今井洋, 八木俊樹, 岩本裕之

    (社)日本動物学会第71回関東支部大会  ( 中央大学理工学部 )   2019年3月  日本動物学会関東支部

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    記述言語:日本語   会議種別:ポスター発表  

    微小管内にはチューブリン分子が、結晶のように整然とラセン状格子配置している。これまで抗がん剤として、種々の微小管安定化剤が開発されて来ているが、微小管構造の安定性とチューブリンの分子構造変化とを関連づけて調べる良い方法がなかった。我々はX線繊維回折法を用いて、タキソールとラウリマライドの2種の安定化剤の効果の明確な違いを明らかにできたので報告する。両者ともに秒単位で微小管構造の変化を引き起こすことがわかったが、チューブリン分子の縦・横方向の相互作用に関して異なる効果を示すことも明らかとなった。

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  • ラフィド藻シャトネラの遊泳停止と赤潮発生機構

    中原 美奈, 成田 大樹, 和田 祐子, 鈴木 雄大, 田中 昂輝, 今井 洋, 上村慎治

    (社)日本動物学会第71回関東支部大会  ( 中央大学理工学部 )   2019年3月  日本動物学会関東支部

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    記述言語:日本語   会議種別:ポスター発表  

    シャトネラ(Chattonella marina var. obata)は、栄養塩類や海水温の条件が整うと急速に増殖・集積する赤潮の原因となるラフィド藻である。小型シャーレ内に培養した高密度のシャトネラを静置すると、数分内に自発的な集積を開始し、時間経過と共に集団で移動する生物対流現象を示すことがわかった。これが実験室内で再現される赤潮現象であると考えられる。この現象は細胞の密度を上げることで促進され、海水中のCa2+濃度を減らすことで抑制されることがわかった。位相差顕微鏡を使ったシャトネラの遊泳行動観察から、細胞密度を上昇させると互いにぶつかり合う頻度が増加し、その機械的な刺激によって短時間の鞭毛打停止反応が起こり、同時に数秒間遊泳停止することがわかった。この鞭毛打停止反応は海水中のCa2+濃度を減らすことで抑制されることもわかった。ここで発見された鞭毛停止反応が、赤潮における細胞集積反応の1つの重要な要因になっていると考えられる。

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  • タコノマクラの作る緑色色素の分泌機構と役割

    加藤佑亮, 中村洋輝, 鬼頭玲賀, 河野美都子, 中原美奈, 上村慎治

    (社)日本動物学会第71回関東支部大会  ( 中央大学理工学部 )   2019年3月  日本動物学会関東支部

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    記述言語:日本語   会議種別:ポスター発表  

    タコノマクラ(Clypeaster japonicus)は、KCl神経刺激や殻部損傷によって緑色の色素を体外へ分泌するため、この色素は生体防御機構と深く関わっていると考えられる。その色素の特性は、Goodwin & Fox(1955)の吸収スペクトルに関する報告しかない。本研究で緑色色素の特性を詳細に調べた結果、親水性の高い高分子量の成分であることがわかり、他種のウニから採取されるナフトキノン系の赤色色素であるechinochromeやspinochrome等とは大きく異なる点、さらに、スカシカシパンにも共通する特性の色素があることがわかった。4腕幼生でもすでに生成機能が備わっており、腕部の機械的な刺激により内胚葉組織付近で分泌される点、その分泌にAchが関わっていることもわかった。

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  • キンギョ精子の運動活性化について

    加木下原自, 奥野誠, 上村慎治

    (社)日本動物学会第71回関東支部大会  ( 中央大学理工学部 )   2019年3月  日本動物学会関東支部

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    記述言語:日本語   会議種別:ポスター発表  

    魚類の精子は、生殖器内と異なる環境に暴露されことで運動が誘発される。今回、キンギョの精子ではイオンや糖の種類にかかわらず、240 mOsmより高い浸透圧で運動が停止することがわかり、運動停止に関しては、K+濃度よりも、高浸透圧が重要であることが示唆された。また、精子の細胞膜を除去し再活性化を試みたところ、ATPが存在すればCaCl2やcAMPを添加していない溶液にても精子の再活性化がみられ、Ca2+、cAMPともに、精子運動開始の細胞内シグナル伝達において必須でない可能性が示唆された。

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  • Biochemical and structural characterization of taxoid microtubule-stabilizing agents

    Estevez-Gallego J, Kamimura S, Balaguer-Perez, F, Lucena-Agell D, Diaz J-F

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018年5月 

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    記述言語:英語   会議種別:ポスター発表  

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  • Structural basis for the GTP activation of tubulin

    Dlaz, Jose-Fernando

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018年5月 

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    記述言語:英語   会議種別:ポスター発表  

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  • Dynamic changes in microtubule structure observed on a time scale of seconds by X-ray fiber diffraction

    Kamimura S, Imai H, Yagi T, Iwamoto H

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018年5月 

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    記述言語:英語   会議種別:ポスター発表  

    Microtubules are key components of the cytoskeleton in eukaryotic cells. Their dynamic conversions between assembled microtubules and free tubulin dimers in cytoplasm are precisely controlled along with the states of cell activities. One of the most fundamental questions of us is how the microtubule dynamics and its structural properties are related to the chemical states of tubulin dimers during GTP-hydrolysis, with the existence of other MAPs and tubulin-binding chemicals.|rn||rn| X-ray fiber diffraction is one of the most powerful techniques to collect the structural information of native microtubules in physiological solutions without fixation, crystallization or freezing. In the present study, we examined the structural changes of microtubules on a time scale of seconds by combining our original technique for shear-flow alignment of microtubules [Sugiyama et al, 2009; Oiwa et al., 2009; Kamimura et al., 2016] with a high-flux beam line at SPring-8 (BL40XU). Our observations clearly showed that binding with paclitaxel induced the decrease of mean microtubule diameter as well as the increase of axial tubulin repeat within a second, indicating configuration changes of tubulin di

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  • Conformational switching of microtubule and cooperative binding of kinesin-1 for polarized transport

    Shima Tomohiro

    EMBO Symposium: Microtubules from Atom to Complex Systems  2018年5月 

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    記述言語:英語   会議種別:ポスター発表  

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  • Biochemical and structural characterization of taxoid microtubule-stabilizing agents

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018年 

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  • Structural basis for the GTP activation of tubulin

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018年 

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  • Dynamic changes in microtubule structure observed on a time scale of seconds by X-ray fiber diffraction

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018年 

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  • Conformational switching of microtubule and cooperative binding of kinesin-1 for polarized transport

    EMBO Symposium: Microtubules from Atom to Complex Systems  2018年 

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  • Conformational switching of microtubule and cooperative binding of kinesin‐1 as a base for polarized transport (B115)

    Shima T, Morikawa M, Kaneshiro J, Kambara T, Kamimura S, Yagi T, Iwamoto H, Uemura S, Shigematsu H, Ichimura T, Watanabe TM, Nitta R, Okada Y, Hirokawa N

    ASCB(米国細胞生物学会)  2017年12月 

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    記述言語:英語   会議種別:ポスター発表  

    Kinesin-1, the founding member of kinesin superfamily proteins, is known to use only a subset of microtubules for transport in living cells. This biased use of microtubules is proposed as the guidance cue for polarized transport in neurons, but the underlying mechanisms are still poorly understood. Here we report that there is a positive feedback in the binding of kinesin-1 to the GDP-microtubule, which spontaneously produces high affinity microtubules from other low-affinity microtubules. This high affinity state requires the binding of kinesin-1 in the nucleotide-free state. Microtubules return to the initial low affinity state by washing out the binding kinesin-1 or by the binding of AMPPNP to kinesin-1. X-ray fiber diffraction, fluorescence speckle microscopy and second harmonic generation microscopy, as well as cryo-EM, collectively demonstrated that the binding of nucleotide-free kinesin to GDP-microtubule changes the conformation of GDP-microtubule to a conformation close to the GMPCPP-microtubule.

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  • 高粘度海水中のマダコ精子鞭毛で顕著な“double wave”の解析

    和田祐子, 最上善広, 上村慎治

    日本動物学会  2017年9月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

    マダコは体内受精と体外受精の中間的な生殖戦略を取る。交尾は行わず、オスは精莢と呼ばれるカプセル入りの精子をメスに手渡し、精子はメスの体表上で精莢から放出され、その後、貯精嚢へと移動する。メスの貯精嚢内で精子は数週間保存され適宜使われる。単離した精莢から海水中に放出された精子を通常海水中で観察すると、頭部を折り曲げその場で回転する運動と、頭部を振って前進する運動とを繰り返しながら絡み合って束を形成する。マダコ精子は約330 μmの長い鞭毛をもち、通常海水中で鞭毛の屈曲は根元と先端では顕著だが、根元から100-200 μmの中間部分ではあまり見られない。海水中に分子モーターダイニンの阻害剤であるシリオブレビン(文献1)を加えると、シリオブレビンAでは波形変化は見られなかったがシリオブレビンDでは後半部分が極端に大きく屈曲した(日本動物学会第86回新潟大会)。|rn| 2%メチルセルロース(1500 cPs)を加えて粘度を上げた海水中で

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  • プラナリアにおける腹部繊毛の形態観察

    日本動物学会 第88回年次大会(富山)  2017年9月 

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    記述言語:日本語   会議種別:ポスター発表  

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  • “Double wave” of Octopus sperm flagella under high viscous condition

    Annual Meeting of Zoological Society of Japan  2017年 

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  • Conformational switching of microtubule and cooperative binding of kinesin‐1 as a base for polarized transport (B115)

    ASCB-EMBO Meeting 2017  2017年 

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    会議種別:ポスター発表  

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  • Flagellar wave form of Octopus vulgaris sperm in high viscosity medium

    Yuuko Wada, Shinji Kamimura

    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan  2016年11月 

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    記述言語:英語   会議種別:ポスター発表  

    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan|rn||rn||rn||rn|Reproduction strategy of octopus is just the intermediate between internal and external fertilization. During mating, male octopus deliver pieces of spermatophore to females. Spermatozoa, after being released from the spermatophores placed on the body surface of females, move or swim towards the spermathicae, sperm storage organs in female oviducts. In spite of detailed descriptions so far on such unique behaviors of Octopus mating, there has been few reports investigating the sperm physiology of Octopus vulgaris. Their flagellar length was about 330 μm with a head of about 25 μm long. We found the manner of flagellar bend propagation was different from usual flagellar motions. It seemed to be propagating along the entire length of flagellar shaft in a non-continuous manner, i.e., smooth bends propagated along the proximal region of ca. 100 μm and then diminished at a middle part of the flagellum (100-200 μm from the base) where flagellum look almost straight, then flagellar bends reappeared around 200 μm from the base. We also found there was difference in the wave

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  • Effects of high pressure on the motility of sea urchin sperm flagella

    Hiroshi Imai, Masayoshi Nishiyama, Yoshie Harada, Shinji Kamimura

    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan  2016年11月 

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    記述言語:英語   会議種別:ポスター発表  

    Sea urchin spermatozoa released from males swim towards eggs in seawater for fertilization in the intertidal zone where sea urchins live. There are many other species living in the deep sea, or some of them accidentally may fall down to the deep sea by gravity after storm. Thus, the fertilization could be occurring under the stress of high or various different pressures. However, there are no report describing how the sperm swimming and flagellar motility is affected under such high pressure conditions.|rn|In order to understand the effects of pressure on motility of sea urchin spermatozoa, we examined sperm motility of sea urchin, Anthocidaris crassispina, under high pressure of seawater in the presence or in the absence of 10 mM CaCl2. Without Ca2+, consistently with previous studies, we observed the circular swimming path of sea urchin sperm on the glass surface under the condition of atmospheric pressure. The path of circular swimming was similar to that of spermatozoa observed in the seawater containing 10 mM CaCl2. We interpreted that the pumping activity of Ca2+ channels or other transporting systems (e.g. Na+/Ca2+ exchanger) can maintain the intracellular Ca2+ concentration

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  • 軸糸直径サイズ変化による鞭毛繊毛の屈曲運動の制御

    Toshiki Yagi, Shinji Kamimura, Hiroyuki Iwamoto

    日本生物物理学会 第54回年会  2016年11月 

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    記述言語:日本語   会議種別:ポスター発表  

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  • Direct flow analysis around the beating flagella of sea-urchin sperm cells supporting the slender body theory of micro-swimmers

    Miyashiro D, Wada Y, Shihira-Ishikawa I, Miya-waki A, Kamimura S

    31st International Congress on High-speed Imaging and Photonics  2016年11月 

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    記述言語:英語   会議種別:口頭発表(一般)  

    For the efficient chemotaxis and fertilization, it should be a crucial problem for animal spermatozoa to swim in the medium without disturbing the concentration gradient of chemical attractants derived from unfertilized eggs. Precise analysis and description of micro-flows occurring around swimming sperm cells was expected to help us to understand the chemotaxis phenomena more in details, however, there were many technical prob-lems to be solved. In the present study, using an in-situ storage image sensor (ISIS) equipped on a differential interference contract (DIC) microscope with a high-intensity xenon-lamp illumination, we successfully record-ed clear images of swimming sea-urchin sperm cells at 6,000 fps in a medium containing micro-particles with 0.1 μm diameter to directly visualize medium flows around bending flagella. We processed the acquired DIC images and traced precise motions of sperm flagella as well as moving free micro-particles with a 0.3-ms time-resolution.|rn|By comparing with theoretical flows expected from the slender body theory (SBT), we concluded that ob-served flows around micro-swimmers such as beating flagella under microscopes are classified as stokes f

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  • 高圧力顕微鏡法による深海微生物の遊泳運動観察

    Masayoshi Nishiyama, Chiaki Kato, Hiroshi Imai, Shinji Kamimura, Yoshie Harada

    日本生物物理学会 第54回年会  2016年11月 

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    記述言語:日本語   会議種別:ポスター発表  

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  • 微小管内GDP-チューブリンの精密な周期決定

    上村慎治, 今井洋, 八木俊樹, 島知弘, 岡田康志, 岩本

    第54回 日本生物物理学会  2016年11月 

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    記述言語:日本語   会議種別:ポスター発表  

    The axial repeat of tubulin molecules in microtubules is roughly 4 nm and has conventionally been used as one of the most convenient standard scales to estimate molecular or organelle size for electron microscopy. However, in our previous report, the axial repeat varied depending on experimental conditions, e.g., temperature, with microtubule stabilizer and GTP-hydrolysis states. It was also shown that the tubulin axial repeat in porcine brain GTP-microtubules, which are mainly composed of GDP-tubulin, was almost constant with a low coefficient of thermal expansion of 3.7×10-5/degree. Here, we determined the accurate repeat of GDP-tubulin molecules after careful revision of camera length, X-ray wavelength as well as the angles of specimen, beam and detector tilting.

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  • X線繊維回折で解き明かす微小管構造の動態

    第87回 日本動物学会(第22回 国際動物学会)  2016年11月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

    X線繊維回折法には、生体試料中の周期構造を直接反映したの回折信号を、< 0.1nmもの高精度でリアルタイム解析できる点で、他手法にはない優れた利点がある。本研究では、一般に重合・脱重合の平衡状態を繰り返すために、動的な構造ゆらぎも大きいと想像される微小管を使った解析結果を紹介する。GTP加水分解や微小管安定化剤添加による構造変化が見られることがわかったが、通常のポリマーには見られない特性、負の熱膨張特性が観察されることがわかった。細胞骨格の示すのユニークな特性について議論したい。

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  • キネシンによる微小管の構造変化

    Tomohiro Shima, Manatsu Morikawa, Junichi Kaneshiro, Taketoshi Kambara, Shinji Kamimura, Toshiki Yagi, Hiroyuki Iwamoto, Taro Ichimura, Tomonobu Watanabe, Sotaro Uemura, Ryo Nitta, Yasushi Okada, Nobutaka Hirokawa

    日本生物物理学会 第54回年会  2016年10月 

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    記述言語:日本語   会議種別:ポスター発表  

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  • X線繊維回折で探る微小管構造の動態

    大阪大学蛋白質研究所セミナー 第6回分子モーター討論会「分子モーター研究の最前線」  2016年7月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

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  • X-ray fiber diffraction analysis of microtubule revealing the dynamic changes of axial tubulin repeats in native microtubules

    S. Kamimura, Y. Fujita, Y. Wada, T. Yagi, T. Shima, Y. Okada, H. Iwamoto

    2016 EMBL Symposia Microtubules, Microtubules: From Atoms to Complex Systems  2016年5月 

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    記述言語:英語   会議種別:ポスター発表  

    One of the most fundamental questions on the molecular mechanisms of microtubule assembly-disassembly dynamics is how the structural stability of microtubules is correlated with the variation of molecular conformation of tubulin dimers within microtubules. To address this issue, we applied our technique for the rapid shear-flow alignment of biological filaments in a physiological buffer medium, enabling us to acquire the structural periodicity data from native microtubules by X-ray fiber diffraction under solution conditions. We could classified microtubules into three main groups on the basis of distinct mean axial tubulin repeats and microtubule diameters that varied depending on GTP hydrolysis and the content of paclitaxel. The paclitaxel effects to induce the elongation of the mean axial repeat of tubulin was observed to be completed within 30 s. It was also suggested that this elongation effects were appearing in a cooperative manner from the observation of microtubules with subthreshold paclitaxel content. Another interesting feature we found was the variation in the temperature dependency of axial tubulin repeat in GTP-, GMPCPP- and GTPγS-microtubules with/without paclitaxe

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  • X線繊維回折で探る微小管構造の動態

    大阪大学蛋白質研究所セミナー 第6回分子モーター討論会「分子モーター研究の最前線」  2016年 

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  • キネシンによる微小管の構造変化

    日本生物物理学会 第54回年会  2016年 

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    会議種別:ポスター発表  

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  • 高圧力顕微鏡法による深海微生物の遊泳運動観察

    日本生物物理学会 第54回年会  2016年 

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    会議種別:ポスター発表  

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  • 微小管内GDP-チューブリンの精密な周期決定

    第54回 日本生物物理学会  2016年 

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    会議種別:ポスター発表  

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  • X線繊維回折で解き明かす微小管構造の動態

    第87回 日本動物学会(第22回 国際動物学会)  2016年 

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  • X-ray fiber diffraction analysis of microtubule revealing the dynamic changes of axial tubulin repeats in native microtubules

    2016 EMBL Symposia Microtubules, Microtubules: From Atoms to Complex Systems  2016年 

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    会議種別:ポスター発表  

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  • Flagellar wave form of Octopus vulgaris sperm in high viscosity medium

    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan  2016年 

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  • Effects of high pressure on the motility of sea urchin sperm flagella

    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan  2016年 

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  • Regulation of cilia and flagella bending movements through the change of axoneme diameter

    The 54th Annual Meeting of the Biophysical Society of Japan  2016年 

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  • Direct flow analysis around the beating flagella of sea-urchin sperm cells supporting the slender body theory of micro-swimmers

    31st International Congress on High-speed Imaging and Photonics  2016年 

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  • X線繊維回折法による微小管内チューブリン構造の動態解析

    上村慎治, 岩本裕之

    日本生物物理学会 第53回年会  2015年9月 

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    記述言語:日本語   会議種別:公開講演,セミナー,チュートリアル,講習,講義等  

    Microtubules play a variety of functions in cells. Recent studies using high-resolution cry-electron micrography and X-ray fiber diffraction revealed that multiple conformational states of tubulin give rise to structural polymorphism in microtubule lattice, which may be crucial for diverse physiological functions of microtubules. Multiple conformations of tubulin also underlie dynamic instability of microtubules, where the balance between assembly and disassembly is stochastically switched in a nucleotide-dependent manner. In this session, we introduce the forefront in the field of microtubule, aiming to share new findings and insights about the conformational dynamics of tubulin and microtubule.

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  • 柔軟性に富んだ微小管内GTPγS-チューブリン分子

    Shinji Kamimura, Yosuke Fujita, Yuuko Wada, Tomohiro Shima, Yasushi Okada, Toshiki Yagi, Hiroyuki Iwamoto

    日本生物物理学会 第53回年会(金沢)  2015年9月 

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    記述言語:日本語  

    Our major question is how tubulin states are related to the stability of microtubules. To address the question, we used our novel technique for the quick shear-flow alignment of biological filaments, which enabled us to acquire fine X-ray fiber diffraction signals from native microtubules in a few seconds. We found that microtubules could be classified into three major groups with distinct axial periodicities of tubulin, which varied depending on the temperature of solution, the state of GTP-hydrolysis and the contents of microtubule stabilizers. It was also revealed that the GTPγS-tubulin showed the widest variation in the longitudinal tubulin periodicity in situ, suggesting a highly flexible state of GTP-tubulin in the microtubules.

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  • 柔軟性に富んだ微小管内GTPγS-チューブリン分子

    日本生物物理学会 第53回年会(金沢)  2015年 

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  • Microtubule dynamics revealed by the X-ray fiber diffraction analysis

    Biophysical Society of Japan. #53 Annual meeting  2015年 

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  • X線繊維回折による鞭毛内チューブリン周期の精密解析

    上村慎治, 岩本裕之

    6th International Symposium on Aero-aqua Bio-Mechanisms (ISABMEC2014)  2014年11月 

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    記述言語:日本語   会議種別:ポスター発表  

    Tubulin is one of the major polypeptides that construct the axonemes of cilia/flagella. Thus, the mechanical or chemical properties of tubulin in axonemes would be optimized for the bending motion or against mechanical stresses given by external liquid. In the present study, with an X-ray fiber diffraction technique, the length changes or variations of tubulin dimer periodicity within microtubules (MT) was investigated with a high precision (~ 0.01 nm). We found there are two main states of tubulin periodicity within usual porcine brain MTs (intracellular singlet MTs), i.e., short and long configurations with a length change by about 3% depending on temperature and taxol binding (MT stabilizer). However, the tubulin repeat within sea-urchin sperm flagella showed just the intermediate of the two states of brain MTs. Here we postulate a new hypothesis, the mechanical capacity of tubulin molecules inside axonemes, which would serve a certain allowance of bending compression-decompression inside MTs.

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  • プラナリア滑走運動における繊毛・筋運動協調関係

    杉田玄太郎, 上村慎治

    6th International Symposium on Aero-aqua Bio-Mechanisms (ISABMEC2014)  2014年11月 

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    記述言語:日本語   会議種別:ポスター発表  

    The gliding motion of flatworms (Planarian, Platyhelminthes) is primarily generated by the effective strokes of ventral cilia. The maximum motion rate of ca. 3-5 mm/s appears to be one of the fastest rates reported for the ciliary motility. Our assumption is there would be fine optimization or regulation of the motile mechanisms to accomplish such a high gliding performance. In the present study, we used different types of specific motility blockers, blebbistatin/BDM for muscular myosin II and ciliobrevin for axonemal dyneins. Blebbistatin blocked peristaltic motions and depressed the half-cylindrical shapes of flatworm body (making flatworms more planar). BDM also blocked gliding motions. We also fund ciliobrevin completely inhibited the gliding motion in minutes, and induced peristaltic shape changes. From our observations, it was suggested that both muscle contraction and ciliary motion, some coordination between them, would be required for the efficient motion of gliding.

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  • 溶液中で配向させたコラーゲンのX 線繊維回折

    Yasunobu Sugimoto, Sakurako Hayashi, Sayaka Hayashi, Nobuhisa Watanabe, Shinji Kamimura, Takanori Kihara

    2014年9月 

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    記述言語:日本語   会議種別:ポスター発表  

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  • ウニ精子鞭毛打運動におけるCiliobrevin D の阻害効果

    和田祐子, 馬場昭次, 上村慎治

    日本動物学会  2014年9月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

    ウニ精子にダイニン特異的な低分子阻害剤cliobrevin A(HPI4)を投与すると鞭毛打周波数が濃度依存的に低下し、高濃度では根元の非対称性が大きな運動となることを前回大会で報告した。今回、合成アナログのciliobrevin D をウニ精子に投与したときの阻害効果をciliobrevin A と比較した。周波数への効果はciliobrevin A と同様だが、波形への効果として鞭毛全長にわたり運動非対称性がさらに増し、鞭毛が頭部の周りに巻きつくものが多数みられた。ダイニン種の違いによる阻害効果の差という観点で考察を加える。

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  • タキソールは、急速な微小管内チューブリン周期の伸長を誘導する

    藤田洋介, 中澤亘將, 岩本裕之, 上村慎治

    日本生物物理学会  2014年9月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

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  • 剪断流下におけるDNA二重ラセン構造のダイナミクス

    藤田洋介, 中澤亘將, 岩本裕之, 上村慎治

    日本生物物理学会  2014年9月 

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    記述言語:日本語   会議種別:ポスター発表  

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  • Paclitaxel添加直後の微小管構造変化

    清原恵, 中澤亘將, 藤田洋介, 和田祐子, 八木俊樹, 岩本 裕之

    日本動物学会  2014年9月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

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  • ミドリムシにおける光運動制御マシナリーの解明

    Shimabukuro, SAP, ミドリムシにおける光運動制御マシナリーの解明|rn|Molecular machinery regulating photomovement of Euglena|rn, 岩崎 憲, 宮崎 直幸, 伊関 峰生, 長谷川 浩司, 成田 哲博

    日本生物物理学会  2014年9月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

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  • ミドリムシにおける光運動制御マシナリーの解明

    日本生物物理学会  2014年 

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  • プラナリア滑走運動における繊毛・筋運動協調関係

    6th International Symposium on Aero-aqua Bio-Mechanisms (ISABMEC2014)  2014年 

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  • X-ray diffraction study of aligned collagen fiber

    2014年 

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  • Inhibition of ciliobrevin D on motility of sea urchin sperm flagella

    2014年 

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  • Dynamic changes of the structure of double helix DNA in shear flow

    2014年 

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  • Length variations of tubulin molecules within native axonemal microtubules

    6th International Symposium on Aero-aqua Bio-Mechanisms (ISABMEC2014)  2014年 

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  • Dynamic changes in axial tubulin repeats in native microtubules revealed by X-ray fiber diffraction

    Dynein 2013 International Workshop (神戸)  2013年10月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • 微小管X 線繊維回折:チューブリンピッチの動的変化

    Shinji Kamimura, Yosuke Fujita, Yuuko Wada, Hiroyuki Iwamoto

    日本製物物理学会第51回年会(京都)  2013年10月 

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    記述言語:日本語   会議種別:ポスター発表  

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  • 微小管のX 線繊維回折:温度、GTP 加水分解、taxol 結合で変わる微小管内のチューブリン構造

    上村慎治, 藤田洋介, 和田祐子, 岩本裕之

    公益社団法人 日本動物学会 第84回大会(岡山)  2013年9月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

    溶液内でtubulin dimerは、curved/straight の2 状態をとると考えられている。微小管内でのtubulin の動的構造変化は、微小管の安定性に大きく影響すると考えられるが、まだ、その詳細を調べた研究報告はない。X 線微小管繊維回折の解析から、taxol 濃度、温度、GTP 加水分解に依存してtubulin ピッチが変わることがわかった。微小管内でtubulin dimer がshort/long の2 状態をとる可能性が考えられる。

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  • ウニ精子鞭毛運動におけるciliobrevinの効果

    和田祐子, 上村慎治

    公益社団法人 日本動物学会 第84回大会  2013年9月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

    細胞質ダイニンの特異的な阻害剤であるciliobrevin の軸糸ダイニンに対する効果を調べた。この物質は低分子で膜透過性を持つので、除|rn|膜していないウニ精子に直接投与した場合に効果を見ることが期待された。予想通り、ciliobrevin は濃度依存的にウニ精子の鞭毛打周波数を下げ、このとき波形の変化も観察された。つまり、内腕、外腕ダイニンの両方に効果があることが示された。

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  • ウニ精子鞭毛運動におけるciliobrevinの効果

    公益社団法人 日本動物学会 第84回大会  2013年 

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  • 微小管X 線繊維回折:チューブリンピッチの動的変化

    日本製物物理学会第51回年会(京都)  2013年 

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  • Xray fiber diffraction of microtubules: Tubulinpitch variations depending on medium temperature, GTPhydrolysis and taxol binding indicating dynamic

    The 84th Annual Meeting of Zoological Society of Japan  2013年 

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  • Dynamic changes in axial tubulin repeats in native microtubules revealed by X-ray fiber diffraction

    Dynein 2013 International Workshop (Kobe)  2013年 

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  • Direct imaging of the world of low Reynolds number hydrodynamics

    D. Miyashiro, Y. Wada, I. Shihira-Ishikawa, A. Miyawaki, S. Kamimura

    The Fifth International Symposium on Aero Aqua Bio-mechanisms, ISABMEC 2012 Taipei  2012年8月 

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    記述言語:英語   会議種別:口頭発表(一般)  

    To clarify the fluid dynamics of swimming microorganisms, a technique to visualize fluid flows under an optical microscope with appropriate spatial and temporal resolution is required. In the present study, we used sea-urchin spermatozoa that have long flagella with planar beat patterns and are suited for precise beat form analysis. Differential inter ference microscopy (DIC) images of the beating sperm flagella and particles included in the medium were recorded with a rate of 6,000 fps. As has been theoretically expected, spermatozoa were observed to swim in a stable medium without perturbation of surrounding fluids. This is the first direct evidence to show how fluid flows around whipping flagella under the condition of low Reynolds number.

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  • 微小管の非等方的冷却収縮

    上村慎治, 岩本裕之

    第10回 日本生物物理学関東支部会  2012年3月  日本生物物理学会

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    記述言語:日本語   会議種別:口頭発表(一般)  

    タンパク分子の構造は,けっして均一で等方的なものではないために,冷却時には非等方的な収縮を示すと予測される。そのような物性はタンパク分子の機能や構造の安定性を理解する上で重要な知見となるが,直接調べることは難しかった。本研究では,流動配向させたウシ脳微小管を15秒で37から17℃まで急速冷却し,同時にX線繊維回折法により構造変化を追跡することに成功した。GDPチューブリンからなる微小管は,低温で脱重合する直前まで,長軸方向・直径方向に,それぞれ60,220×10-6/Kの熱膨張係数を示し,非等方的な冷却収縮を示すことがわかった。

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  • Direct imaging of the world of low Reynolds number hydrodynamics

    The Fifth International Symposium on Aero Aqua Bio-mechanisms, ISABMEC 2012 Taipei  2012年 

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  • Mechanical properties of flagellar motility: Prokaryotes versus Eukaryotes

    上村慎治

    FASEB Summer Research Conference, Saxtons River, Vermont  2010年7月 

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    記述言語:英語   会議種別:ポスター発表  

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  • A new function of primary cilia: a cell-signaling enhancer

    高尾大輔, 上村慎治

    FASEB Summer Research Conference, Saxtons River, Vermont  2010年7月 

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  • Mechanical properties of flagellar motility: Prokaryotes versus Eukaryotes

    FASEB Summer Research Conference, Saxtons River, Vermont  2010年 

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  • A new function of primary cilia: a cell-signaling enhancer

    FASEB Summer Research Conference, Saxtons River, Vermont  2010年 

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  • Mechanical properties of flagellar motility: Prokaryotes versus eukaryotes.

    International Workshop Dynein 2009, 2009.11.3, Kobe(Japan)  2009年11月 

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    記述言語:英語   会議種別:ポスター発表  

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  • X-ray fiber diffraction analysis of axoneme and microtubules structure using a novel shear-flow fiber-aligning technique.

    上村慎治

    International Workshop Dynein 2009, 2009.11.3, Kobe(Japan)  2009年11月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • Mechanical properties of flagellar motility: Prokaryotes versus eukaryotes.

    4th International Symposium on Aero Aqua Bio-Mechanisms., 2009.8.30, Shanghai(China)  2009年8月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • Mechanical properties of flagellar motility: Prokaryotes versus eukaryotes.

    4th International Symposium on Aero Aqua Bio-Mechanisms., 2009.8.30, Shanghai(China)  2009年 

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  • Mechanical properties of flagellar motility: Prokaryotes versus eukaryotes.

    International Workshop Dynein 2009, 2009.11.3, Kobe(Japan)  2009年 

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  • X-ray fiber diffraction analysis of axoneme and microtubules structure using a novel shear-flow fiber-aligning technique.

    International Workshop Dynein 2009, 2009.11.3, Kobe(Japan)  2009年 

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  • A new microscope optics for laser darkfield illumination applied to high precision measurement of specimen displacement.

    Noda, N, Kamimura, S

    Biophys. Soc. 52nd Ann. Meeting/ 16th IUPAB Int. Biophys. Congress/Biophysical Society  2008年3月 

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    記述言語:英語   会議種別:口頭発表(一般)  

    With conventional light microscopy, precision in the measurement of the displacement of a specimen depends on the signal-to-noise ratio when we measure the light intensity of magnified images. This implies that, for the improvement of precision, getting b

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  • Dynein arm arrangement in sea-urchin sperm flagellar axonemes revealed by small-angle X-ray diffraction analysis.

    Kamimura, S, Iwamoto, H

    Biophys. Soc. 52nd Ann. Meeting/ 16th IUPAB Int. Biophys. Congress/Biophysical Society  2008年3月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • A new microscope optics for laser darkfield illumination applied to high precision measurement of specimen displacement.

    Biophys. Soc. 52nd Ann. Meeting/ 16th IUPAB Int. Biophys. Congress/Biophysical Society  2008年 

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  • Dynein arm arrangement in sea-urchin sperm flagellar axonemes revealed by small-angle X-ray diffraction analysis.

    Biophys. Soc. 52nd Ann. Meeting/ 16th IUPAB Int. Biophys. Congress/Biophysical Society  2008年 

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  • Dynamic change of axonemeal structure and dynein arm arrangement in sea-urchin sperm flagella revealed by small-angle X-ray diffraction analysis.

    Kamimura, S, Iwamoto, H

    FASEB Summer Research Conference (The Biology of Cilia & Flagella)/FASEB organization  2007年8月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • Dynein arm arrangement in flagellar axonemes and its dynamic change revealed by small-angle X-ray diffraction analysis.

    Kamimura S, Iwamoto, H

    The Molecular Motor Conference/Fujihara Foundation of Science  2007年8月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • Dynamic change of axonemeal structure and dynein arm arrangement in sea-urchin sperm flagella revealed by small-angle X-ray diffraction analysis.

    FASEB Summer Research Conference (The Biology of Cilia & Flagella)/FASEB organization  2007年 

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  • Dynein arm arrangement in flagellar axonemes and its dynamic change revealed by small-angle X-ray diffraction analysis.

    The Molecular Motor Conference/Fujihara Foundation of Science  2007年 

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  • X-ray diffraction from dyenin motors and microtubules in sea -urchin sperm flagellar axonemes.

    Kamimura, S

    生物物理,日本生物物理学会  2006年11月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • 3-D measurement of the nanometer displacement of microspheres under optical mixcroscope

    Noda, N, Kamimura, S

    生物物理,日本生物物理学会  2006年11月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • 新しい流動配向法によるウニ精子鞭毛X線回折の観察

    上村 慎治, 杉山 貴紹, 杉本 泰伸, 若林克三

    社団法人日本動物学会第77回大会,日本動物学会  2006年9月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

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  • X-ray diffraction from dyenin motors and microtubules in sea -urchin sperm flagellar axonemes.

    2006年 

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  • 3-D measurement of the nanometer displacement of microspheres under optical mixcroscope

    2006年 

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  • 鞭毛軸糸高速微小振動のナノメーター計測

    野田直紀, 上村慎治

    生物物理,日本生物物理学会  2005年11月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

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  • AFMを用いたウニ精子鞭毛軸糸における高速微小振動の検出

    榊原肇

    生物物理,日本生物物理学会  2001年10月 

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  • 急峻なATP濃度上昇に伴うウニ精子除膜鞭毛モデルの運動

    谷 知己, 上村慎治

    生物物理,日本生物物理学会  1996年11月 

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    記述言語:日本語   会議種別:口頭発表(一般)  

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▼全件表示

Works(作品等)

  • 在外研究

    2018年7月    

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    "21 shifts/7 days at BM26/DUBBLE, 27 shifts/9 days at BL11/NCD-SWEET, 6 days at D16/ILL"

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  • 在外研究

    2018年7月 -  

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  • 在外研究

    2017年10月    

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    12 shifts/4 days at BM26/DUBBLE

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  • 在外研究

    2017年10月 -  

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  • X線繊維回折法による微小管動態の高速追跡

    2016年4月 -  

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  • Analysis of microtubule structure dynamics by X-ray fiber diffraction.

    2016年4月 -  

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  • 超高分解能光学顕微鏡の開発と応用

    2009年4月 -  

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  • 3次元ピコメートル計測法による軸糸ダイニン動態の解析

    2007年8月 -  

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  • 新配向技術によるベン毛軸糸X線構造解

    2007年4月 -  

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  • Study on the axonemal structure using a novel fiber aligning technique.

    2007年4月 -  

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▼全件表示

受賞

  • 藤井賞・Zoological Science論文賞

    2011年9月   日本動物学会   Single-Cell Electroporation of Fluorescent Probes into Sea Urchin Sperm Cells and Subsequent FRAP Analysis

    高尾大輔, 上村慎治

  • ZOOLOGICAL SCIENCE Award 2011

    2011年  

  • 井上研究奨励賞

    1985年12月   井上科学振興財団  

    上村慎治

共同研究・競争的資金等の研究課題

  • 微小管内チューブリン分子ラセン配置の柔軟性と可塑性

    2019年4月 - 2021年3月

    基盤研究(C)(一般) 

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    資金種別:競争的資金

    配分額:4290000円 ( 直接経費:4290000円 、 間接経費:990000円 )

    微小管は、真核生物のさまざまな細胞機能に深く関わる細胞骨格である。シンクロトロンの高輝度X線源を使ったX線繊維回折法を使って、微
    小管構造の柔軟性・可塑性を正確に定量的な評価を行うことで、微小管の生理機能をより深く理解できると共に、抗がん剤としての微小管安定
    化剤の薬理効果を客観的に数値化できる利点がある。これまでの電子顕微鏡技術ではわからなかった情報、動的な構造変化、その変化の時定数
    、構造の熱ゆらぎの評価を行う新手法として確立することで、抗がん剤の網羅的な探索研究を推進する。

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  • 変異チューブリンによる重合―GTP加水分解の共役メカニズムの解明

    研究課題/領域番号:17H03668  2017年4月 - 2020年3月

    日本学術振興会  科学研究費助成事業  基盤研究(B)  国立研究開発法人理化学研究所

    武藤 悦子, 今井 洋, 上村 慎治

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    配分額:17030000円 ( 直接経費:13100000円 、 間接経費:3930000円 )

    微小管のダイナミクスは細胞分裂や形態形成に重要な役割を果たしている。GTPチューブリンからどのように重合核が作られ微小管になるか、その仕組みはこれまでほとんど明らかにされていなかった。我々はラピッドフラッシュネガティブ染色電子顕微鏡法と反応速度論的解析を組み合わせることにより、微小管の核生成には、GTPチューブリンが集合して直線型oligomerが作られる必要があることを見出した。直線型oligomerが伸びて一定の長さ(臨界核)に達すると、ラテラルな相互作用によりmulti-stranded oligomerとなり、そこからプロトフィラメントが安定に重合するようになることがわかった。

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  • X線繊維回折法による微小管動態の高速追跡

    2016年4月 - 2019年3月

    文部科学省  科学研究費補助金(日本学術振興会・文部科学省) 

    上村慎治

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    資金種別:競争的資金

    配分額:3800000円

    "当研究室で独自に開発し、応用実験へも成功した流動配向技術+X線繊維回折法(Sugiyama et al.,|rn|2009; Kamimura et al., in press)に、微少量液体噴出制御技術を組み合わせ、水溶液中で薬剤添加直|rn|後の微小管の急速な構造変化を調べる新手法として完成させる。この新手法を微小管構造安定性|rn|に影響を与える薬剤の網羅的検索へと応用すると同時に、イオン濃度変化、温度、pH 変化などの|rn|物理化学的な溶液条件で急速に(<5 s)変化する微小管内のチューブリン分子構造の動態、および、|rn|格子配置における協同性についての分子的な基盤を明らかにする。"

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  • 細胞骨格動態の温度特性解析

    2015年 - 2018年

    基礎科学研究 

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    資金種別:競争的資金

    哺乳類脳微小管は、非常に高い温度感受性を有する点で、他の細胞骨格とは大きく性質を異にしている。また、GTP加水分解状態によっては、負の熱膨張係数(NTE)を持つ点も、当研究室の仕事から明らかになってきた。溶液条件で、この温度特性がどのように変化するのがを解明することで、微小管の特異的な低温感受性の生物学的な意味を理解することを目指す。

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  • Temperature dependent dynamics of cytoskeletal components in eukaryote cells.

    2015年 - 2018年

    Basic Science Research Program 

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    資金種別:競争的資金

    Microtubules are key components of the cytoskeleton in eukaryotic cells. The dynamics between assembled microtubules and free tubulin dimers in the cytoplasm is closely related to the active shape changes of microtubule networks. One of the most unique features of microtubules is its quite high sensitivity to lowered temperatures. Mammalian brain microtubules disassemble in second by lowering the medium temperature below 20ºC. Other interesting properties of microtubules are negative-thermal expansion depending on the condition of GTP-hydrolysis by tubulin dimers. To understand the detailed mechanism of such temperature-sensitivity, we are analyzing the conformation changes of tubulin-molecules within microtubule by X-ray fiber diffraction.

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  • チューブリン・微小管の動的構造解析を利用した抗ガン作用物質の網羅的検索

    2014年 - 2018年

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    資金種別:競争的資金

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  • Efficient survey of anti-cancer drugs by a novel X-ray fiber diffraction technique to analyze dynamic configuration changes of tubulin molecules within native microtubules.

    2014年 - 2018年

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    資金種別:競争的資金

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  • X線繊維回折法を使った微小管構造に対する諸薬剤の効果

    2012年 - 2017年

    ライフサイエンス基礎科学研究 

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    資金種別:競争的資金

    微小管を安定化させるPaclitaxel(taxol)は、通常、微小管内のtubulin dimerに作用し、protofilament間の結合を強めることによって、微小管の脱会合が抑えられると考えられる。これまでの研究から、tubulinの縦方向の周期が長くなり、平均protofilament数が徐々に減少するとの報告もある。本研究では、BL45XU-SAXS(SPring-8)でのX線繊維回折法によって、構造の経時変化を分単位で追跡することで、動的な構造変化がわかった。

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  • Tubulin fine structures in native microtubules revealed by X-ray fiber diffarction

    2012年 - 2017年

    Basic Science Research on Life Science 

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    資金種別:競争的資金

    Microtubules (MT) are key components of the cytoskeleton in eukaryotic cells. One of the most undamental questions is how MT dynamics is associated with the molecular conformation of tubulin within MTs. To address the issues, we applied our new technique for the rapid shear-flow alignment of biological filaments, which enabled us to acquire X-ray fiber diffraction data in seconds from oriented native MTs under various physiological conditions. We found that the mean periodicity of tubulin dimers in MTs were elongated from 3.85 to 3.95 nm in 1 min. Diameter changes appeared to be occurring in a 10-20 min timecourse. It is suggested paclitaxel induces quick elongation of tubulin dimers in MTs.

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  • 3次元ピコメートル計測法による軸糸ダイニン動態の解析

    2007年8月 - 2009年3月

    文部科学省  科学研究費補助金(日本学術振興会・文部科学省)-特定領域研究 

    上村慎治

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    資金種別:競争的資金

    配分額:5000000円

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  • 生理学的な条件下での鞭毛・微小管の構造解析

    2007年 -  

    ライフサイエンス基礎科学研究 

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    資金種別:競争的資金

    新規の流動配向法を開発し、生体の繊維構造(微小管・軸糸・アクチン繊維)の動的な変化がはじめて追跡できるようになった。この手法で、これまで不明であった生体繊維の機能解明を目指している。

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  • Structure analysis of axonemes and microtub ule under physiological conditions.

    2007年 -  

    Basic Science Research on Life Science 

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    資金種別:競争的資金

    Our novel technique to align biological fiber in a physiological buffer medium enabled us to start new investigation on the dynamic chnages of various biological fibers. We are now revealing new features of biologically important fiber structures.

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  • 精密マイクロフロー解析による微小生物遊泳機序の研究

    研究課題/領域番号:13450096  2001年 - 2004年

    日本学術振興会  科学研究費助成事業  基盤研究(B)  東京大学

    上村 慎治, 高野 泰斉, 小林 俊一, 須田 斎, 篠原 健一

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    配分額:13000000円 ( 直接経費:13000000円 )

    低レイノルズ数条件下での生物遊泳を解析・模倣することで、新しいデザインのマイクロマシンを創出することを目標に研究を行った。上村は、光学センサーを使い、運動を定量化する装置を開発し、ニジマスやウニ精子の運動解析、微環境の粘度計測に応用した。また、非常に薄い水層で精子の運動を観察する新手法を開発した。高野は、マイクロフローのシミュレーションを手がけた。ベン毛断面の捩りおよび曲げモーメントを求め、Kirchhoff-Rod理論からベン毛の形状変化を予測し、実験結果との比較からビブリオ歯ベン毛の曲げ剛性を評価した。また、サルモネラ菌ベン毛の素繊維がR/Lの2状態を取ることを考慮した形状予測も行った。ストークス流の細長物体理論をベン毛に、境界要素法を菌体に適用することで細菌遊泳運動の解析も行った。小林は、ベン毛運動の屈曲メカニズムを模倣した拡大モデルの開発、および、コンピュータシミュレーションを行った。シミュレーションでは、流体内推進機構の弾性や周囲液体の粘性の推力や速度へ影響を検討した。拡大モデルについては柔軟体にダイニンに相当する電磁石やギヤ付モーターを取り付けて屈曲動作を発生させ,流体内での推力を検討した。須田は、駆動に関わるモーター分子の作動機構の解明を目指した。麦面自由エネルギー勾配のある面上を液滴が滑ることを実験的に示した。また、ミオシン分子の構造変化と力発生の同時計測を行った。物質表面の示す吸着力の荷重依存性を調べ、生体分子モーターとの比較検討や表面間力-結合寿命間の相関を調べた。篠原は、次世代の機能性材料と期待されるπ共役系高分子に着目し、SPMやAFM技術を高分子化学の分野へ新たに取り込んだ。高分子鎖1本の観察に成功し、ポリマー鎖のミクロブラウン運動に基づくスペクトルの動的挙動とポリマー鎖1本の運動を直接観測するのに成功し、基礎化学の分野に大きなインパクトを与えた。

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  • 生物分子モーターの1分子計測と1分子操作

    研究課題/領域番号:09279104  1997年 - 2000年

    日本学術振興会  科学研究費助成事業  特定領域研究(A) 

    上村 慎治, 木下 一彦, 嶋本 伸雄, 柳田 敏雄, 木下 一彦, 大岩 和弘, 上田 太朗, 上村 慎治, 上田 太郎

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    配分額:195400000円 ( 直接経費:195400000円 )

    モーター分子の動態を一分子レベルで解析する新手法を開発し応用研究を行った。木下は分子方向性を調べる新手法でミオシン上を滑走するアクチンの回転を測定した。柳田はミオシン一分子を観測する技術でATP加水分解反応と滑り運動との間に数100msの遅れがあることを発見した。上村は、一分子FRETを実時間観測できる装置を開発し、総合班との共同研究でミオシン一分子のFRET変動を観測した。嶋本は、大腸菌trpリプレッサーがDNA上をスライドし特異的部位に対する親和性を高めることを明らかにした。モーター分子の動態は、大きな目印を結合させて解析する手法も極めて有効である。木下は、F1-ATPaseに蛍光アクチンを付ける画期的な方法で120度の回転ステップを持ち無負荷条件では毎秒100回転の高速となることを示した。柳田は、微小ガラス針にミオシン分子を結合させる実験系で一分子の力〜速度関係や速度の温度依存性は筋線維と同じであることを示した。本プロジェクトでは一分子解析に将来結びつく可能性の高い新材料・新手法の開発にも着手した。上村はcagedATPを使った解析で力発生とdynein・ADP生成過程とが合致することを示した。上田は、単頭ミオシンとの比較研究より双頭分子のモーター領域間には機能的共同性があることやG680V変異ミオシンは安定な架橋を形成することなどを示した。また、大岩はクラミドモナスのdynein cは8nmのステップ変位を持ち連続的な運動を起こすこと、dynein fはduty ratioの高いモーターであることなどを発見した。嶋本はRNAポリメラーゼの転写開始期に短鎖RNAのみを合成する転写中間体があり、その生成はポリメラーゼ同士の衝突で促進されることを発見した。新しい蛍光性ATPアナログを用いミオシン結合に伴う蛍光変化を見る手法(大岩)、高精度のAFM装置でミオシンフィラメントの負電荷量分布を調べる研究(柳田)、分子モーターを使い人工的に一方向へ運動させる手法の開発(上田)、AFMによるモーター分子振動現象の観測(上村)など、今後応用的な面で期待される新手法も多く試みた。

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  • 生物分子モーターの多様性と同一性:総括班

    研究課題/領域番号:09279101  1997年 - 2000年

    日本学術振興会  科学研究費助成事業  特定領域研究(A)  東京大学

    須藤 和夫, 豊島 陽子, 神谷 律, 上村 慎治, 大沢 文夫, 江橋 節郎, 木下 一彦

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    配分額:126600000円 ( 直接経費:126600000円 )

    細胞体の移動、細胞分裂、細胞内輪送、分泌、食作用など、真核生物の「動き」には、ミオシン、キネシン、ダイニンなどの生物分子モーターが深く関わっている。生物分子モーターはヌクレオチドに蓄えられた化学エネルギーを力や滑り運動に変換するという共通の機能を持つものの、その機能には驚くべき多様性がある。(1)1分子計測・操作を用いて生物分子モーターがもつ機能の多様性をあきらかにするとともに、(2)こうした多様性を生み出す構造的基盤を構造解析を通してあきらかにすることが、本特定領域研究の主な目標であった。このような目標を達成するため、「生物分子モーターの1分子計測・操作」班、「生物分子モーターの動的構造解析」班、「生物分子モーターの探索」班という3つの計画研究班を置くとともに公募研究を募った。総括班は、こうした大きな組織を有機的につなぐ要として機能するため、班会議あるいは海外からの研究者もふくめた国際シンポジウムを毎年開催し、また計測センターを設置するなどして、計画、公募班内あるいは班間での共同研究を活発化することに努めた。この結果、数多くの論文がいくつかのグループの共同研究として出版され、本研究を成功させることができた。

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  • 我が国の動物学の効果的推進に関する総合的研究

    研究課題/領域番号:08304046  1996年 - 1998年

    日本学術振興会  科学研究費助成事業  基盤研究(B)  筑波大学

    岡田 益吉, 上村 慎治, 神崎 亮平, 七田 芳則, 川村 和夫, 安部 眞一

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    配分額:5100000円 ( 直接経費:5100000円 )

    (1)1998年5月23日に米子(動物・植物・生態三学会中四国支部大会)で、「第2回科研費フォーラム」を開催した。「科学研究助成費の日米比較」をテーマとして、林正男氏(お茶の水女子大学理学部助教授)と宮嶌和男氏(文部省学術国際局研究 助成課助成企画室長)に講演していただいた。(2)4月27日「日本の動物学の推進のために」という提言を動物学会理事会に提出した。学生・ボスドク・教職会員への啓蒙活動を行うこと、動物学会大会の活性化に向けて本大会の課題と改革すべき点があること、学会内外での充分な広報活動を可能にするために、広報委員会を設置すること、留学生や在日研究者の研究活動を支援し、アジアの動物学を振興すること、などの提言を行った。(3)7月23日に高知で、広報委員会の具体的形態と、科研費の審査体制と評価体制、について議論した。また、広島本大会で開催する予定の第3回科研費フォーラムについて話し合った.
    (4)9月26日広島の動物学会本大会時に科研費フォーラムを開き、谷口直之氏(大阪大学医学部教授、日本生化学会副会長)、遠藤啓氏(日本学術振興会総務部長、前 研究助成課課長)、増本健氏(東北大学名誉教授、電気磁気材料研究所所長、日本学術会議会員)の3氏をパネリストとして招待し、「審査員の人数」、「プロジェクトの規模」、「審査結果の開示」のテーマに絞って会場との討論を行った。
    (5)1999年1月22〜23日に京都で委員会を開き、第3期将来計画委員会の総括を行い、第4期に引き継ぐ問題点について議論した。科研費問題については、文部省に対する提言をまとめるべく議論した。

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  • ピコメータ精度計測システムを使った蛋白分子間相互作用の解析法

    研究課題/領域番号:07558096  1995年 - 1997年

    日本学術振興会  科学研究費助成事業  基盤研究(A)  東京大学

    上村 慎治

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    配分額:13600000円 ( 直接経費:13600000円 )

    本研究では、精密測定を目的とした水平式の光学系を持つ光学顕微鏡システムの開発に成功した。光学系を水平式とする利点は、光軸を安定して固定できるために機械的に極めて安定した装置ができる点、対物レンズと接眼レンズ間の距離を自由に変更できるため拡大率を調節可能な点、光学素子を支えるための鏡筒部分が不要なため対物・接眼の間に自由に他の部品、たとえば、UV光学系、ダイクロイックミラー、シャッターなどを挿入できる点、観察試料のマイクロマニピュレーションを行うために装置が設置しやすい点など多くのものがある。また計測装置の光センサを時間分解能の高い設定に改良することで時間分解能が10KHzまで向上できるように改良した。この装置を使ってウニ精子微小管の振動波形を詳しく解析した結果、振動には一定速度の滑り運動を行う相(A)と周期的なゆらぎのみられるプラトー相(B)に分けることができることがわかった。Aはダイニンとチューブリンの強い相互作用(架橋)が生じている状態、Bは、その架橋が解離してATPの再結合までの状態ではないかと考えられる。また、この装置を用い、以下のcaged-ATPを使った新しい実験を試みた。トリトン処理したウニ精子鞭毛を実験材料として、瞬間的(数ミリ秒以内)にATPを付与し、その時の微小管滑り運動をnm〜数百pmの精度で計測することができた。その結果、鞭毛軸糸の中のダイニンがATPと結合し、ダイニン・ADPの反応中間体となったものが、滑り力発生(運動の開始)に関与している可能性が最も高いことが証明できた。これはこれまでテトラヒメナを使って行われてきたダイニンATP加水分解酵素に関する生化学的な報告を裏付ける結果であり、ダイニンのin situでの酵素化学反応論的な実測としては、はじめての報告である。反面、ダイニン・ADPと急速に結合するために強い阻害効果を持つと考えられて来たバナジン酸は、この運動開始に関わっていると考えられるダイニン・ADP反応中間体には全く作用しないということもこの研究で判明した。力発生のメカニズムを理解する上で非常に重要な知見である。

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  • nm〜pm精度実時間運動解析技術を用いたキネシン滑り運動機構の研究

    研究課題/領域番号:06044064  1994年    

    日本学術振興会  科学研究費助成事業  国際学術研究  東京大学

    上村 慎治, TRINCZEK Ber, MANDELKOW Ec

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    配分額:5200000円 ( 直接経費:5200000円 )

    キネシンはATPをエネルギー源として微小管にそった滑り運動を担う運動性蛋白質である。はじめは神経軸索輸送にあずかる酵素として発見されたが細胞内輸送機構や細胞分裂など他の重要な機能にも関係した蛋白質であることがわかってきた。おなじような運動性蛋白質であるダイニン(鞭毛繊毛や神経軸索に存在)やミオシン(筋肉)と同じように運動のメカニズムは不明な点が多い。近年の遺伝子工学的な手法の発達にともない分子のアミノ酸1次配列を人為的に変えて運動性に重要な影響を与える配列を調べてゆくという研究が現在各国で始まりつつある。このような研究の進展にともない機能を詳細に確かめる手段,つまり,運動を分子スケールの精密さで解析する手段がますます必要になってきている。この研究の目的は,2つの技術をすでに実用化している日独2つの研究室の間の共同研究の形でこの研究を効率的に進め,運動の分子機構を解明することである。
    まず,本研究ではブタ脳から従来の手法で抽出精製されたキネシンと形質転換させた大腸菌から精製し頭部ドメインの340〜390個のアミノ酸(運動活性の中心と考えられている箇所)との間での運動活性を詳細に比較する実験から始めた.この研究は主にドイツ側での実験設備(微分干渉顕微鏡と画像処理装置)を使用して行われた.ここで使用したキネシンの頭部ドメインは微小管結合部位,およびATP結合部位を有するモーター活性ドメインではあるが,そのままでは運動の有無を調べることはできない.これまでの手法ではこの頭部ドメインのC端側あるいはN端側に別のアミノ酸配列を追加し,この部分をマイクロビーズやガラスに付着させる方法がとられている.この手法の欠点は付加するアミノ酸配列の種類によってはモーター活性が失われたり,極めて低い活性しか見られないケースがあり,付加したペプタイドの2次的な影響を無視できない点である.これは頭部ドメインのアミノ酸の中でどの残基が重要であるかを今後調べて行くとき実験結果の解釈上の難点となる.可能な限り別のアミノ酸配列を付加させずに運動活性を調べる手法をまず開発することが研究をすすめる上で重要である.
    本研究で試みた手法は,ブタ脳から精製されたキネシンと大腸菌キネシン頭部ドメインとのインビトロ競合実験である.大腸菌キネシンの濃度を上昇させることでブタ脳キネシンによる運動速度は減少し,大腸菌キネシンの頭部ドメインの速度に漸近してゆくことがわかった.特にキネシン頭部ドメインN端側340個アミノ酸のペプタイドでは毎秒約0.05μmの速度となり,この部位がブ脳キネシンの約1/10の運動活性を示すことが示唆された.この手法のコンピュータシミュレーション結果との比較,別のアミノ酸配列ペプタイドの比較は今後の重要課題であると考えられる.特に重要な働きを持つと考えられるアミノ酸残基を入れ替えたとき,簡便にその運動活性の有無を検査できる手法として利用できる可能性がある.
    上のような実験と平行してキネシンと微小管との相互作用を計測する装置の開発を主として日本側の研究室で行ってきた.nm〜pmの精度で運動の解析をおこなう上でのもっとも重要な課題は,機械的な振動ノイズに対する対策と光学顕微鏡の改良である。光学ベンチなどの除振台は数Hz以上の振動を防ぐ手段としてはきわめて有効であるが,より低い周波数の振動が問題になることが多い。本研究では光学顕微鏡下の極めて微小な運動を計測し解析できる新しい光学顕微鏡計測システムを試作した.市販の光学顕微鏡は通常のプレパラート観察のために操作性を重視した設計となっているために本研究の様な精度を達成するには不向きである.そのため新しく光学系を設計することから研究を開始した.レンズ系を簡略化した水平式の光学顕微鏡とすることで上の目的に適した装置を完成させることができた.また,レーザー光源の使用による計測精度の千倍以上の向上,顕微操作技術との組み合わせなど改良も行った.この新しく開発された装置を使用し微小管関連タンパク質であるタウタンパク質と微小管との相互作用,とくに微小管の上でどのようなブラウン運動をしているのか,結合解離のダイナミックな変化を生理的な条件下で調べる実験を現在進めている.

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  • 鞭毛軸系高速振動現象における力学・化学カップリングの解析

    研究課題/領域番号:04740399  1992年    

    日本学術振興会  科学研究費助成事業  奨励研究(A)  東京大学

    上村 慎治

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    配分額:1000000円 ( 直接経費:1000000円 )

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  • 滑り運動の多様性と統一性

    研究課題/領域番号:01657001  1989年    

    日本学術振興会  科学研究費助成事業  重点領域研究  名古屋大学

    神谷 律, 豊島 陽子, 茶圓 茂, 真行寺 千佳子, 上村 慎治, 今栄 康雄

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    配分額:22000000円 ( 直接経費:22000000円 )

    本研究は生体運動の共通原理を探るという立場から、細菌鞭毛、真核細胞鞭毛、原形質流動など筋肉以外の生体運動の機構を追及した。方法として、運動に関与した構造一つづつの動態を直視することを重視した。豊島は単離した骨格筋の太い繊維上をアクチン繊維が滑走するという試験管内運動系を開発し、アクチン繊維が双極性のミオシン繊維のどちらの半分とでも相互作用して滑ることができるという現象を観察した。これはアクチンとミオシンの相互作用にはこれまでの想像を超えた自由度があることを意味している。今栄は細菌鞭毛モーターの共有結果性阻害剤を開発し、その存在下で1個のモーターの挙動を追跡したところ、回転運動が段階的に阻害されていくことを見いだした。運動に関与したイオンチヤンネル1つづつが阻害されていく過程を捉えたものと考えられる。真行寺は真核生物の鞭毛に外力を加えてその応答を見る研究を行った。その結果、鞭毛の打つ面を人為的に回転させることができることを発見した。微小管滑り運動の調節という観点からきわめて興味深い現象である。上村は空間精度1nm、時間分解能1msecで運動を検出できる装置を開発し、上村と神谷はそれによって鞭毛軸糸内の微小な構造の揺らぎを検出する試みを行った。その結果、一見運動を停止している軸糸が、ATP存在下で、振幅1nm、周期300Hzという高速微小振動を行う現象を発見した。その振動数は個々のダイニンの動きを反映している可能性がある。今後、この現象の意味が明らかになれば、運動の分子機構の解明に大いに役立つ実験系になるであろう。この方法をアクチンーミオシン系など他の生体運動系に適用し、微少な運動の揺らぎをさまざまな条件下で検出/解析することにより、化学ー力学変換過程に関する新情報が得られる可能性がある。現在そのような試みが茶圓によって準備されている。来年度からの成果が待たれる。

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  • ベン毛などの運動の制御機構・基本構造・力学的な特性が生物進化でどんな意味付けができるのか探っています。

    1984年 -  

    基礎科学研究 

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    資金種別:競争的資金

    真核生物の鞭毛・繊毛の運動の制御機構・基本構造・力学的な特性を調べ、この運動機構が、生物進化の上でどのような意味付けができるのか探っています。特に、真核生物は何故すべてが鞭毛・繊毛を持つに至ったのか(その子孫であるのか)、その利点と欠点を理解することで、真核生物の誕生の過程が解明できると考えています。

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  • Motility mechanisms and mechanical properties of eukaryote falagellum and its evolutional aspects.

    1984年 -  

    Basic Science Research Program 

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    資金種別:競争的資金

    We are now investigating the mechanisms of regulation, structural and mechanical properties of eukaryotic cilia and flagella, expecting such studies would lead us to understand the evolutionary history of eukaryotes. A major question is why all the eukaryotes (or our ancient organisms) had to have cilia and flagella. We can know the answer to this question after the fundamental understanding of the advantage and disadvantage of the motile system of cilia and flagella.

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  • 2009年 -  

    (社)日本動物学会   評議員