Updated on 2024/02/15

写真a

 
FUKUI Akimasa
 
Organization
Faculty of Science and Engineering Professor
Other responsible organization
Biological Sciences Course of Graduate School of Science and Engineering, Master's Program
Biological Sciences Course of Graduate School of Science and Engineering, Doctoral Program
Contact information
The inquiry by e-mail is 《here
External link

Degree

  • 博士(理学) ( 横浜市立大学 )

  • 理学修士 ( 横浜市立大学 )

Education

  • 1994.3
     

    Yokohama City University   Graduate School, Division of Integrated Science   doctor course   completed

  • 1991.3
     

    Yokohama City University   Graduate School, Division of Integrated Science   master course   completed

  • 1989.3
     

    Yokohama City University   graduated

  • 1984.3
     

    神奈川県立港南台高等学校   graduated

Research History

  • 2017.4 -  

    中央大学理工学部 教授

  • 2010.4 - 2017.3

    北海道大学大学院先端生命科学研究院先端融合科学研究部門 准教授

  • 2006.4 - 2010.3

    北海道大学大学院理学研究院生命理学専攻 准教授

  • 2005.10 - 2006.3

    北海道大学大学院理学院生命理学部門 助教授

  • 1994.8 - 2005.9

    東京大学大学院総合文化研究科生命環境系 助手

  • 1994.4 - 1994.7

    日本学術振興会特別研究員

▼display all

Professional Memberships

  •   - Now

    Society of Evolutinary Studies, Japan

  •   - Now

    公益社団法人 日本動物学会

  •   - Now

    日本発生生物学会

  •   - Now

    特定非営利活動法人 日本分子生物学会

Research Interests

  • gastrulation

  • macrophages

  • regeneration

  • cell migration

  • allotetraploid genome

  • morphogenesis

Research Areas

  • Life Science / Genome biology  / Genome biology

  • Life Science / Cell biology  / 細胞生物学

  • Life Science / Developmental biology  / 発生生物学

Papers

  • Gene Structure Analysis of Chemokines and Their Receptors in Allotetraploid Frog, Xenopus laevis Reviewed International journal

    Fukui, A, Matsunami, M

    Frontiers in Genetics   12   787979 - 787979   2022.1

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Frontiers  

    Chemokines, relatively small secreted proteins, are involved in cell migration and function in various biological events, including immunity, morphogenesis, and disease. Due to their nature, chemokines tend to be a target of hijacking of immunity by virus and therefore show an exceptionally high mutation rate. Xenopus laevis is considered an excellent model to investigate the effect of whole-genome duplication for gene family evolution. Because its allotetraploidization occurred around 17-18 million years ago, ancestral subgenomes L and S were well conserved. Based on the gene model of human and diploid frog Xenopus tropicalis, we identified 52 chemokine genes and 26 chemokine receptors in X. laevis. The retention rate of the gene in the X. laevis L and S subgenomes was 96% (45/47) and 68% (32/47), respectively. We conducted molecular phylogenetic analysis and found clear orthologies in all receptor genes but not in the ligand genes, suggesting rapid divergences of the ligand. dN/dS calculation demonstrated that dN/dS ratio greater than one was observed in the four ligand genes, cxcl8b.1.S, cxcl18.S, ccl21.S, and xcl1.L, but nothing in receptor genes. These results revealed that the whole-genome duplication promotes diversification of chemokine ligands in X. laevis while conserving the genes necessary for homeostasis, suggesting that selective pressure also supports a rapid divergence of the chemokines in amphibians.

    DOI: 10.3389/fgene.2021.787979

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  • A comprehensive reference transcriptome resource for the Iberian ribbed newt Pleurodeles waltl, an emerging model for developmental and regeneration biology. Reviewed International journal

    Masatoshi Matsunami, Miyuki Suzuki, Yoshikazu Haramoto, Akimasa Fukui, Takeshi Inoue, Katsushi Yamaguchi, Ikuo Uchiyama, Kazuki Mori, Kosuke Tashiro, Yuzuru Ito, Takashi Takeuchi, Ken-Ichi T Suzuki, Kiyokazu Agata, Shuji Shigenobu, Toshinori Hayashi

    DNA research : an international journal for rapid publication of reports on genes and genomes   26 ( 3 )   217 - 229   2019.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OXFORD University press  

    Urodele newts have unique biological properties, notably including prominent regeneration ability. The Iberian ribbed newt, Pleurodeles waltl, is a promising model amphibian distinguished by ease of breeding and efficient transgenic and genome editing methods. However, limited genetic information is available for P. waltl. We conducted an intensive transcriptome analysis of P. waltl using RNA-sequencing to build and annotate gene models. We generated 1.2 billion Illumina reads from a wide variety of samples across 12 different tissues/organs, unfertilized egg, and embryos at eight different developmental stages. These reads were assembled into 1,395,387 contigs, from which 202,788 non-redundant ORF models were constructed. The set is expected to cover a large fraction of P. waltl protein-coding genes, as confirmed by BUSCO analysis, where 98% of universal single-copy orthologs were identified. Ortholog analyses revealed the gene repertoire evolution of urodele amphibians. Using the gene set as a reference, gene network analysis identified regeneration-, developmental-stage-, and tissue-specific co-expressed gene modules. Our transcriptome resource is expected to enhance future research employing this emerging model animal for regeneration research as well as for investigations in other areas including developmental biology, stem cell biology, and cancer research. These data are available via our portal website, iNewt (http://www.nibb.ac.jp/imori/main/).

    DOI: 10.1093/dnares/dsz003

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  • A simple culture method for liver and intestinal tissue-resident macrophages from neonatal mice. Reviewed

    Shimizu, Y, Sakuragi, N, Nakamura, K, Taira, T, Ayabe, T, Fukui, A

    In Vitro Cellular & Developmental Biology - Animal   55 ( 6 )   436 - 444   2019.5

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer  

    DOI: 10.1007/s11626-019-00359-y

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    Other Link: http://link.springer.com/article/10.1007/s11626-019-00359-y/fulltext.html

  • Clustered Xenopus keratin genes: A genomic, transcriptomic, and proteomic analysis Reviewed

    Ken-ichi T. Suzuki, Miyuki Suzuki, Mitsuki Shigeta, Joshua D. Fortriede, Shuji Takahashi, Shuuji Mawaribuchi, Takashi Yamamoto, Masanori Taira, Akimasa Fukui

    DEVELOPMENTAL BIOLOGY   426 ( 2 )   384 - 392   2017.6

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Keratin genes belong to the intermediate filament superfamily and their expression is altered following morphological and physiological changes in vertebrate epithelial cells. Keratin genes are divided into two groups, type I and II, and are clustered on vertebrate genomes, including those of Xenopus species. Various keratin genes have been identified and characterized by their unique expression patterns throughout ontogeny in Xenopus laevis; however, compilation of previously reported and newly identified keratin genes in two Xenopus species is required for our further understanding of keratin gene evolution, not only in amphibians but also in all terrestrial vertebrates. In this study, 120 putative type I and II keratin genes in total were identified based on the genome data from two Xenopus species. We revealed that most of these genes are highly clustered on two homeologous chromosomes, XLA9_10 and XLA2 in X. laevis, and XTR10 and XTR2 in X. tropicalis, which are orthologous to those of human, showing conserved synteny among tetrapods. RNA-Seq data from various embryonic stages and adult tissues highlighted the unique expression profiles of orthologous and homeologous keratin genes in developmental stage- and tissue-specific manners. Moreover, we identified dozens of epidermal keratin proteins from the whole embryo, larval skin, tail, and adult skin using shotgun proteomics. In light of our results, we discuss the radiation, diversification, and unique expression of the clustered keratin genes, which are closely related to epidermal development and terrestrial adaptation during amphibian evolution, including Xenopus speciation.

    DOI: 10.1016/j.ydbio.2016.10.018

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  • Genome evolution in the allotetraploid frog Xenopus laevis. Reviewed

    Session, A.M, Uno, Y, Kwon, T, Chapman, J.A, Toyoda, A, Takahashi, S, Fukui, A, Hikosaka, A, Suzuki, A, Kondo, M, van Heeringen, S.J, Quigley, I, Heinz, S, Ogino, H, Ochi, H, Hellsten, U, Lyons, J.B, Simakov, O, Putnam, N, Stites, J, Kuroki, Y, Tanaka, T, Michiue, T, Watanabe, M, Bogdanovic, O, Lister, R, Georgiou, G, Paranjpe, S.S, van Kruijsbergen, I, Shu, S, Carlson, J, Kinoshita, T, Ohta, Y, Mawaribuchi, S, Jenkins, J, Grimwood, J, Schmutz, J, Mitros, T, Mozaffari, S, Suzuki, Y, Haramoto, Y, Yamamoto, T.S, Takagi, C, Heald, R, Miller, K, Haudenschild, C, Kitzman, J, Nakayama, T, Izutsu, Y, Robert, J, Fortriede, J, Burns, K, Lotay, V, Karimi, K, Yasuoka, Y, Dichmann, D.S, Flajnik, M.F, Houston, D.W, Shendure, J, DuPasquier, L, Vize, P.D, Zorn, A.M, Ito, M, Marcotte, E, Wallingford, J.B, Ito, Y, Asashima, M, Ueno, N, Matsuda, Y, Veenstra, G.C, Fujiyama, A, Harland, R.M, Taira, M, Rokhsar, D.S

    NATURE   538 ( 7625 )   336 - 343   2016.9

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  • SDF-1 alpha regulates mesendodermal cell migration during frog gastrulation Reviewed

    Akimasa Fukui, Toshiyasu Goto, Junko Kitamoto, Motohiro Homma, Makoto Asashima

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   354 ( 2 )   472 - 477   2007.3

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    During frog gastrulation, mesendodermal cells become apposed to the blastocoel roof (BCR) by endoderm rotation, and migrate towards the animal pole. The leading edge of the mesendodermal cells (LEM) contributes to the directional migration of involuting marginal zone (IMZ) cells, but the molecular mechanism of this process is not well understood. Here we show that CXCR4/SDF-1 signaling mediates the directional movement of the LEM in Xenopus embryos. Expression of xCXCR4 was detected in the IMZ, and was complemented by xSDF-1 alpha expression in the inner surface of the BCR. Over-expression of xCXCR4 and xSDF-1 alpha caused gastrulation defects. An xCXCR4 N-terminus deletion construct and xSDF-l alpha-MO also inhibited gastrulation. Furthermore, explants of LEM migrate towards the dorsal BCR in the presence of xSDF-1a, and altered xCXCR4 expression in the LEM inhibited LEM migration. These results suggest that CXCR4/SDF-1 signaling is necessary for the migrations of massive numbers of cells during gastrulation. (c) 2007 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2007.01.007

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  • IDENTIFICATION OF ACTIVIN-A, ACTIVIN-AB, AND ACTIVIN-B AND FOLLISTATIN PROTEINS IN XENOPUS-EMBRYOS Reviewed

    A FUKUI, T NAKAMURA, H UCHIYAMA, K SUGINO, H SUGINO, M ASASHIMA

    DEVELOPMENTAL BIOLOGY   163 ( 1 )   279 - 281   1994.5

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS  

    There are several lines of evidence that activin is a crucial molecule for mesoderm induction in early Xenopus embryos. However, it is not known what kind and what amounts of activin proteins exist in cleavage stage embryos. Three isc,forms of activins, A, AB, and B, were demonstrated to be present at least in part as a complex with abundant follistatin, an activin-binding protein, in early Xenopus embryos (stage 1-5). These results suggest that activin stored in eggs has an important role for mesoderm induction during early Xenopus embryogenesis and that follistatin can modulate the function of activin in signaling developmental changes. (C) 1994 Academic Press, Inc.

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  • Sex chromosome differentiation and the W- and Z-specific loci in Xenopus laevis Reviewed

    Shuuji Mawaribuchi, Shuji Takahashi, Mikako Wada, Yoshinobu Uno, Yoichi Matsuda, Mariko Kondo, Akimasa Fukui, Nobuhiko Takamatsu, Masanori Taira, Michihiko Ito

    DEVELOPMENTAL BIOLOGY   426 ( 2 )   393 - 400   2017.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Genetic sex-determining systems in vertebrates include two basic types of heterogamety; XX (female)/ XY (male) and ZZ (male)/ZW (female) types. The African clawed frog Xenopus laevis has a ZZ/ZW-type sex-determining system. In this species, we previously identified a W-specific sex (female)-determining gene dmw, and specified W and Z chromosomes, which could be morphologically indistinguishable (homomorphic). In addition to dmw, we most recently discovered two genes, named scanw and ccdc69w, and one gene, named capn5z in the W-and Z-specific regions, respectively. In this study, we revealed the detail structures of the W/Z-specific loci and genes. Sequence analysis indicated that there is almost no sequence similarity between 278 kb W-specific and 83 kb Z-specific sequences on chromosome 2Lq3233, where both the transposable elements are abundant. Synteny and phylogenic analyses indicated that all the W/Z-specific genes might have emerged independently. Expression analysis demonstrated that scanw and ccdc69w or capn5z are expressed in early differentiating ZW gonads or testes, thereby suggesting possible roles in female or male development, respectively. Importantly, the sex-determining gene (SDG) dmw might have been generated after allotetraploidization, thereby indicating the construction of the new sex-determining system by dmw after species hybridization. Furthermore, by direct genotyping, we confirmed that diploid WW embryos developed into normal female frogs, which indicate that the Z-specific region is not essential for female development. Overall, these findings indicate that sex chromosome differentiation has started, although no heteromorphic sex chromosomes are evident yet, in X laevis. Homologous recombination suppression might have promoted the accumulation of mutations and transposable elements, and enlarged the W/Z-specific regions, thereby resulting in differentiation of the W/Z chromosomes. (C) 2016 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.ydbio.2016.06.015

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  • Conservatism and variability of gene expression profiles among homeologous transcription factors in Xenopus laevis Reviewed

    Minoru Watanabe, Yuuri Yasuoka, Shuuji Mawaribuchi, Aya Kuretani, Michihiko Ito, Mariko Kondo, Haruki Ochi, Hajime Ogino, Akimasa Fukui, Masanori Taira, Tsutomu Kinoshita

    DEVELOPMENTAL BIOLOGY   426 ( 2 )   301 - 324   2017.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Xenopus laevis has an allotetraploid genome of 3.1 Gb, in contrast to the diploid genome of a closely related species, Xenopus tropicalis. Here, we identified 412 genes (189 homeolog pairs, one homeologous gene cluster pair, and 28 singletons) encoding transcription factors (TFs) in the X. laevis genome by comparing them with their orthologs from X. tropicalis. Those genes include the homeobox gene family (Mix/Bix, Lhx, Nkx, Paired, POU, and Vent), Sox, Fox, Pax, Dmrt, Hes, GATA, T-box, and some clock genes. Most homeolog pairs for TFs are retained in two X. laevis subgenomes, named L and S, at higher than average rates (87.1% vs 60.2%). Among the 28 singletons, 82.1% were deleted from chromosomes of the S subgenome, a rate similar to the genome-wide average (82.1% vs 74.6%). Interestingly, nkx2-1, nkx2-8, and pax9, which reside consecutively in a postulated functional gene cluster, were deleted from the S chromosome, suggesting cluster-level gene regulation. Transcriptome correlation analysis demonstrated that TF homeolog pairs tend to have more conservative developmental expression profiles than most other types of genes. In some cases, however, either of the homeologs may show strongly different spatio-temporal expression patterns, suggesting neofunctionalization, subfunctionalization, or nonfunctionalization after allotetraploidization. Analyses of otxl suggests that homeologs with much lower expression levels have undergone greater amino acid sequence diversification. Our comprehensive study implies that TF homeologs are highly conservative after allotetraploidization, possibly because the DNA sequences that they bind were also duplicated, but in some cases, they differed in expression levels or became singletons due to dosage-sensitive regulation of their target genes.

    DOI: 10.1016/j.ydbio.2016.09.017

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  • Xenopus laevis全ゲノム解析 転写因子をコードする遺伝子群の初期発生および成体器官における発現パターンの解析

    渡部 稔, 回渕 修治, 安岡 有理, 伊藤 道彦, 近藤 真理子, 越智 陽樹, 荻野 肇, 福井 彰雅, 平良 眞規, 木下 勉

    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集   88回・38回   [1P0559] - [1P0559]   2015.12

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  • Real-Time Imaging of Actin Filaments in the Zebrafish Oocyte and Embryo Reviewed

    Yumiko Nukada, Mayu Horie, Akimasa Fukui, Tomoya Kotani, Masakane Yamashita

    CYTOSKELETON   72 ( 9 )   491 - 501   2015.9

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-BLACKWELL  

    Dynamic changes of cytoplasmic and cortical actin filaments drive various cellular and developmental processes. Although real-time imaging of actin filaments in living cells has been developed, imaging of actin filaments in specific cells of living organisms remains limited, particularly for the analysis of gamete formation and early embryonic development. Here, we report the production of transgenic zebrafish expressing the C-terminus of Moesin, an actin filament-binding protein, fused with green fluorescent protein or red fluorescent protein (GFP/RFP-MoeC), under the control of a cyclin B1 promoter. GFP/RFP-MoeC was expressed maternally, which labels the cortical actin cytoskeleton of blastula-stage cells. High levels of GFP/RFP fluorescence were detected in the adult ovary and testis. In the ovaries, GFP/RFP-MoeC was expressed in oocytes but not in follicle cells, which allows us to clearly visualize the organization of actin filaments in different stages of the oocyte. Using full-grown oocytes, we revealed the dynamic changes of actin columns assembled in the cortical cytoplasm during oocyte maturation. The number of columns slightly decreased in the early period before germinal vesicle breakdown (GVBD) and then significantly decreased at GVBD, followed by recovery after GVBD. Our transgenic fish are useful for analyzing the dynamics of actin filaments in oogenesis and early embryogenesis. (C) 2015 Wiley Periodicals, Inc.

    DOI: 10.1002/cm.21253

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  • Application of Multichannel Collagen Gels in Construction of Epithelial Lumen-like Engineered Tissues Reviewed

    Kazuya Furusawa, Takeomi Mizutani, Hiromi Machino, Saki Yahata, Akimasa Fukui, Naoki Sasaki

    ACS BIOMATERIALS SCIENCE & ENGINEERING   1 ( 7 )   539 - 548   2015.7

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    Introduction of epithelial lumen-like structures such as blood and lymphatic vessels, as well as renal tubules, is a prerequisite for successful construction and function of artificially engineered giant tissues. Here, we demonstrate a methodology for construction of various epithelial lumen-like structures by using multichannel collagen gels (MCCGs). MCCGs were prepared and used as template scaffolds for constructing epithelial lumen structures in a controlled fashion. The effect of NaCl concentration on the multichannel structure of MCCGs was investigated by using confocal laser scanning microscopy along with fluorescent staining. The channel diameter increased with increasing NaCl concentrations in the collagen solution and the phosphate buffer solution. In contrast, the channel number decreased with increasing NaCl concentrations. Engineered tissues with various lumen-like structures were constructed by seeding and culturing Madin-Darby canine kidney cells on MCCGs. The diameter of the lumen and the number of lumens per unit area were controllable by regulating the multichannel structure of cylindrical MCCG. We believe that our methodology for the construction of engineered tissues possessing epithelial lumen-like structures will prove helpful in regeneration of giant tissues with various hierarchical structures.

    DOI: 10.1021/acsbiomaterials.5b00003

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  • Molecular Role of RNF43 in Canonical and Noncanonical Wnt Signaling Reviewed

    Tadasuke Tsukiyama, Akimasa Fukui, Sayuri Terai, Yoichiro Fujioka, Keisuke Shinada, Hidehisa Takahashi, Terry P. Yamaguchi, Yusuke Ohba, Shigetsugu Hatakeyama

    MOLECULAR AND CELLULAR BIOLOGY   35 ( 11 )   2007 - 2023   2015.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC MICROBIOLOGY  

    Wnt signaling pathways are tightly regulated by ubiquitination, and dysregulation of these pathways promotes tumorigenesis. It has been reported that the ubiquitin ligase RNF43 plays an important role in frizzled-dependent regulation of the Wnt/beta-catenin pathway. Here, we show that RNF43 suppresses both Wnt/beta-catenin signaling and noncanonical Wnt signaling by distinct mechanisms. The suppression of Wnt/beta-catenin signaling requires interaction between the extracellular protease-associated (PA) domain and the cysteine-rich domain (CRD) of frizzled and the intracellular RING finger domain of RNF43. In contrast, these N-terminal domains of RNF43 are not required for inhibition of noncanonical Wnt signaling, but interaction between the C-terminal cytoplasmic region of RNF43 and the PDZ domain of dishevelled is essential for this suppression. We further show the mechanism by which missense mutations in the extracellular portion of RNF43 identified in patients with tumors activate Wnt/beta-catenin signaling. Missense mutations of RNF43 change their localization from the endosome to the endoplasmic reticulum (ER), resulting in the failure of frizzled-dependent suppression of Wnt/beta-catenin signaling. However, these mutants retain the ability to suppress noncanonical Wnt signaling, probably due to interaction with dishevelled. RNF43 is also one of the potential target genes of Wnt/beta-catenin signaling. Our results reveal the molecular role of RNF43 and provide an insight into tumorigenesis.

    DOI: 10.1128/MCB.00159-15

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  • Effects of a flow field on amyloid fibrillogenesis in a beta-lactoglobulin solution Reviewed

    R. K. Sharma, K. Furusawa, A. Fukui, N. Sasaki

    INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES   70   490 - 497   2014.9

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    The effects of a flow field on the amyloid fibrillogenesis of beta-lactoglobulin (beta LG) were investigated using a flow birefringence method and AFM imaging experiments. A 4 wt% beta LG aqueous solution was incubated at pH 2 and 80 degrees C. A flow field was then applied by stirring at 250 and 474 rpm. An incubation without stirring was used as a control sample. Flow birefringence measurements were taken at room temperature from the incubated sample solutions in which an elongational flow field was used. The birefringence pattern obtained indicated that the fibrils formed by the incubation were rigid rod-like molecules. Birefringence relaxation experiments revealed at least two relaxation processes, suggesting a double peaked distribution function for fibrils length. The length distribution of fibrils expected from the birefringence experiments was confirmed by the AFM images of amyloid fibrils. The order of the expected length of the resultant fibrils in both longer and shorter length distributions was those stirred at 250 rpm congruent to 474 rpm > 0 rpm. The effects of the flow field applied during the incubation on amyloid fibrillogenesis was discussed on the basis of the rate process consideration. The present results demonstrated that the flow field should be considered as an important factor that regulates the fibrillogenesis of globular proteins. (C) 2014 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.ijbiomac.2014.06.034

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  • Control of Structure of Multi-Channel Collagen Gel Beads. Reviewed

    Kazuya Furusawa, Akimasa Fukui, Naoki Sasaki

    2014 INTERNATIONAL SYMPOSIUM ON MICRO-NANOMECHATRONICS AND HUMAN SCIENCE (MHS)   1 - 3   2014

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    Language:English   Publishing type:Research paper (international conference proceedings)   Publisher:IEEE  

    To control the structure of collagen gel with multichannel structure (Multi-Channel Collagen Gel: MCCG), the effect of gelation temperature on the multichannel structure of MCCG have been investigated. The diameter of channel decreased with increasing gelation temperature. By contrast, the number of channel increased with increasing the gelation temperature. The results showed that the multi-channel structure of MCCG can be controlled by regulating the gelation temperature.

    DOI: 10.1109/MHS.2014.7006141

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  • Multiscale Analysis of Changes in an Anisotropic Collagen Gel Structure by Culturing Osteoblasts Reviewed

    Yohei Hanazaki, Jyun-ichi Masumoto, Shoichi Sato, Kazuya Furusawa, Akimasa Fukui, Naoki Sasaki

    ACS APPLIED MATERIALS & INTERFACES   5 ( 13 )   5937 - 5946   2013.7

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    Mimicking the complicated anisotropic structures of a native tissue is extremely important in tissue engineering. In a previous study, we developed an anisotropic collagen gel scaffold (ACGS) having a hierarchical structure and a properties gradient. In this study, our objective was to see how cells remodel the scaffolds through the cells-ACGS interaction. For this purpose, we cultured osteoblastic cells on ACGS, which we regarded as a model system for the cells-extracellular matrix (cell-ECM) interaction. Changes in the ACGS-cell composites structure by cell-ECM interactions was investigated from a macroscopic level to a microscopic level. Osteoblastic cells were also cultured on an isotropic collagen gel (ICGS) as a control. During the cultivation, mechanical stimuli were applied to collagen-cell composites for adequate matrix remodeling. Confocal laser scanning microscope (CLSM) was used to observe macroscopic changes in the ACGS-cell composite structure by osteoblastic cells. Small-angle X-ray scattering (SAXS) measurements were performed to characterize microscopic structural changes in the composites. Macroscopic observations using CLSM revealed that osteoblastic cells remained only in the diluted phase in ACGS and they collected collagen fibrils or formed a toroidal structure, depending on the depth from the ACGS surface in the tubular diluted phase. The cells were uniformly distributed in ICGS. SAXS analysis suggests that collagen fibrils were remodeled by osteoblastic cells, and this remodeling process would be affected by the structure difference between ACGS and ICGS. These results suggest that we directly regulate cell-ECM interaction by the unique anisotropic and hierarchical structure of ACGS. The cell-gel composite presented in this study would promise an efficient scaffold material in tissue engineering

    DOI: 10.1021/am303254e

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  • Changes in the viscoelastic properties of cortical bone by selective degradation of matrix protein Reviewed

    Hideki Shirakawa, Kazuya Furusawa, Akimasa Fukui, Shigeru Tadano, Naoki Sasaki

    JOURNAL OF BIOMECHANICS   46 ( 4 )   696 - 701   2013.2

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    We have studied stress relaxation of bovine femoral cortical bone specimens treated with KOH aqueous solution which had been known to degrade selectively protein molecules in bone. With the KOH treatment, we found an increase in specimens' volume. This increase was regarded as swelling of the bone specimen, presumably due to matrix protein network degradation including that of collagen. In an analogy of bone to gel structure, an increasing ratio of specimen volume was used as an indicating parameter for the matrix protein network degradation by the treatment. Although an empirical equation with a linearly combined form of two Kohlrausch-Williams-Watts (KWW) functions has been shown to describe the stress relaxation of bone specimens, a single KWW function was suitable for the bone specimens treated with KOH solution for as little as 3 h. In KOH treated specimens, both the initial modulus and the relaxation time decreased with the volume-increasing ratio, while the relaxation time distribution did not change. A chemo-rheological consideration attributed the reduction of modulus values to the network degradation in the organic matrix phase. The relaxation time of KOH treated specimens was thought to be related to the longer relaxation time of untreated bones, although there was a discontinuity between the extrapolated relaxation time values for KOH treated specimens and untreated specimens. This discontinuity may have originated from the release of residual stress existing in the bone by the matrix protein degradation. The results of the present study suggest that the state of matrix protein is crucial for integrating the mechanical properties of bone. (c) 2012 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.jbiomech.2012.11.038

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  • Change in the viscoelastic properties of agarose gel by HAp precipitation by osteoblasts cultured in an agarose gel matrix Reviewed

    Yohei Hanazaki, Daisuke Ito, Kazuya Furusawa, Akimasa Fukui, Naoki Sasaki

    Journal of Biorheology   26 ( 1-2 )   21 - 28   2013

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    The viscoelastic properties of cell-seeded agarose gel were measured as a function of culture time. Because the seeded cells, MC3T3-E1, were osteoblast-like cells, the system can be regarded as a model osteogenesis system. For all specimens the characteristic stress relaxation curve of agarose gel was observed-a large relaxation up to 104 s followed by a gel plateau, where the former was attributed to molecular motion of polymer chains between two adjacent cross-links of the gel and the latter to the elasticity of the gel network. The viscoelasticity was quantified by fitting stress relaxation data to an empirical equation. The relaxation time and its distribution did not change with culture time. The initial and equilibrium moduli, E0 and Ee, respectively, and relaxation strength, ΔE = E0 - Ee, did not change up to day 15 of culture but changed significantly at day 18 of culture. The change in ΔE with culture period correlated well with that in E0. The changes in the mechanical properties of the cell-seeded agarose gel system were explained in terms of the function of MC3T3-E1 in precipitating calcium phosphate mineral particles. The precipitation was detected by alizarin red S staining of the system at day 9 of culture. The precipitated calcium phosphate was confirmed to be hydroxyapatite (HAp) and the amount of HAp increased monotonically with culturing time, both of which were observed by X-ray diffraction studies. The dependence of the modulus of the composite on mineral fraction is discussed. A simple model of mixing of the components based on the continuum material concept was not applicable, but a model considering percolation of mineral particles in a network chain with culture time was suitable to explain the observed results. The results may be particularly important for predicting the stiffness of functionally engineered bony tissue implanted in a fractured bone. © 2011 Japanese Society of Biorheology.

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  • Expression of xSDF-l alpha, xCXCR4 and xCXCR7 during gastrulation in Xenopus laevis Reviewed

    Surabhi-Kirti Mishra, Tomoko Nagata, Kazuya Furusawa, Naoki Sasaki, Akimasa Fukui

    INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY   57 ( 1 )   95 - 100   2013

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    Chemokines play a crucial role in developmental processes and recent studies have revealed that they also control gastrulation movements. In this paper, we report the expression patterns of xSDF-l alpha, xCXCR4 and xCXCR7 and regulation of the expression of xSDF-la and xCXCR4 during gastrulation. We performed whole mount in situ hybridization (WISH) and quantitative realtime RT-PCR (qRT-PCR) analyses to examine the distribution of transcripts. The effect of activin/nodal signaling on the expression of xSDP-1 alpha and its receptors was examined by animal cap assay and microinjection of cer-s mRNA. We have demonstrated that the xSDF-la transcript is increased in the blastocoel roof during gastrulation, but not in the involuted mesoderm. xCXCR4 was expressed in the mesendoderm at late blastula and was retained throughout gastrulation. xCXCR7 was found in the dorsal lip around the blastopore in the early gastrula stage and became localized in the presumptive notochord later. We also show that the expression of xCXCR4 and xSDF-l alpha were reciprocally regulated by activin/nodal signaling. These results suggest that xSDP-1 alpha and its receptors contribute to the cell arrangement of mesoderm cells and their expression patterns are partially regulated by activin/nodal signaling.

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  • Modulation of cell-cell adhesion in Xenopus mesendoderm cells by CXCR7 signaling Reviewed

    Keita Masuda, Kazuya Furusawa, Naoki Sasaki, Akimasa Fukui

    14th International Xenopus Conference program book   101   2012.9

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  • Studies on the Formation Mechanism and the Structure of the Anisotropic Collagen Gel Prepared by Dialysis-Induced Anisotropic Gelation Reviewed

    Kazuya Furusawa, Shoichi Sato, Jyun-ichi Masumoto, Yohei Hanazaki, Yasuyuki Maki, Toshiaki Dobashi, Takao Yamamoto, Akimasa Fukui, Naoki Sasaki

    BIOMACROMOLECULES   13 ( 1 )   29 - 39   2012.1

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    We have found that dialysis of S mg/mL collagen solution into the phosphate solution with a pH of 7.1 and an ionic strength of 256 mM at 25 degrees C results in a collagen gel with a birefringence and tubular pores aligned parallel to the growth direction of the gel. The time course of averaged diameter of tubular pores during the anisotropic gelation was expressed by a power law with an exponent of 1/3, suggesting that the formation of tubular pores is attributed to a spinodal decomposition-like phase separation. Small angle light scattering patterns and high resolution confocal laser scanning microscope images of the anisotropic collagen gel suggested that collagen fibrils are aligned perpendicular to the growth direction of the gel. The positional dependence of the order parameter of the collagen fibrils showed that the anisotropic collagen gel has an orientation gradient.

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  • Wide Angle X-ray Diffraction and Small Angle X-ray scattering Studies on the Effect of Demineralization on Hierarchical Structure of Bone Reviewed

    Kazuya Furusawa, Hideki Shirakawa, Shoichi Sato, Yuka Tomimori, Bijey Giri, Akimasa Fukui, Naoki Sasaki

    Trans. Mat. Res. Soc. Japan   36 ( 3 )   387 - 391   2011

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  • Xenopus furry contributes to release of microRNA gene silencing Reviewed

    Toshiyasu Goto, Akimasa Fukui, Hiroshi Shibuya, Ray Keller, Makoto Asashima

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   107 ( 45 )   19344 - 19349   2010.11

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    A transcriptional corepressor, Xenopus furry (Xfurry), is expressed in the chordamesodermal region and induces secondary dorsal axes when overexpressed on the ventral side of the embryo. The N-terminal furry domain functions as a repressor, and the C-terminal leucine zipper (LZ) motifs /coiled-coil structure, found only in vertebrate homologs, contributes to the nuclear localization. The engrailed repressor (enR)+LZ repressor construct, which has properties similar to Xfurry, induced several chordamesodermal genes. In contrast, an antisense morpholino oligonucleotide, Xfurry-MO, and the activating construct, herpes simplex virus protein (VP16)+LZ, had effects opposite those of Xfurry overexpression. Because blocking protein synthesis with cycloheximide superinduced several Xfurry transcriptional targets, and because expression of enR+LZ induced such genes under cycloheximide treatment, we analyzed the role of an Xfurry transcriptional target, microRNA miR-15. Cycloheximide reduced the expression of primary miR-15 (pri-miR-15), whereas miR-15 reduced the expression of genes superinduced by cycloheximide treatment. These results show that Xfurry regulates chordamesodermal genes by contributing to repression of pretranscriptional gene silencing by miR-15.

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  • Highly conserved functions of the Brachyury gene on morphogenetic movements: Insight from the early-diverging phylum Ctenophora Reviewed

    Atsuko Yamada, Mark Q. Martindale, Akimasa Fukui, Shin Tochinai

    DEVELOPMENTAL BIOLOGY   339 ( 1 )   212 - 222   2010.3

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    Brachyury, a member of the T-box transcription family identified in a diverse array of metazoans, was initially recognized for its function in mesoderm formation and notochord differentiation in vertebrates: however, its ancestral role has been suggested to be in control of morphogenetic movements. Here, we show that morpholino oligonucleotide knockdown of Brachyury (MlBra) in embryos of a ctenophore, one of the most ancient groups of animals, prevents the invagination of MlBra expressing stomodeal cells and is rescued with corresponding RNA injections. Injection of RNA encoding a dominant-interfering construct of MlBra causes identical phenotypes to that of RNA encoding a dominant-interfering form of Xenopus Brachyury (Xbra) in Xenopus embryos. Both injected embryos down-regulate Xbra downstream genes, Xbra itself and Xwnt11 but not axial mesodermal markers, resulting in failure to complete gastrulation due to loss of convergent extension movements. Moreover, animal cap assay reveals that MlBra induces Xwnt11 like Xbra. Overall results using Xenopus embryos show that these two genes are functionally interchangeable. These functional experiments demonstrate for the first time in a basal metazoan that the primitive role of Brachyury is to regulate morphogenetic movements, rather than to specify endomesodermal fates, and the role is conserved between non-bilaterian metazoans and vertebrates. (C) 2009 Elsevier Inc. All rights reserved.

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  • Effect of mineral dissolution from bone specimens on the viscoelastic properties of cortical Reviewed

    Naoki Sasaki, Tsutomu Nozoe, Ryoji Nishihara, Akimasa Fukui

    JOURNAL OF BIOMECHANICS   41 ( 16 )   3511 - 3514   2008.12

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    Although physiological saline (0.15 M NaCl aqueous solution) has been used as a storage Solution to prevent bone specimens from drying, there have been reports that Ca(2+) ions dissolve front bone specimens during the storage in saline. In order to determine whether Such storage has a marked effect oil mechanical properties, the relaxation Modulus of bovine cortical bone stored in physiological saline was compared with that stored in a buffer solution containing a Sufficient amount of Ca(2+) ions. After storage in saline, the modulus Value of specimens was significantly reduced from that before storage. Oil the other hand, the modulus value of specimens soaked in the solution containing Sufficient Ca(2+) ions did not change after storage. The relaxation rate of a bone specimen stored in physiological saline was larger than that of a specimen stored in Ca(2+)-buffered saline solution and that of the control specimens. The results Suggest that by the dissolution of Ca(2+) ions from a bone specimen during storage in physiological saline, percolated paths of mineral phase and of reinforced matrix phase are disjoined, resulting ill reduction in the elastic modulus and change in the viscoelastic properties of bone. (C) 2008 Elsevier Ltd. All rights reserved.

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  • XHAPLN3 plays a key role in cardiogenesis by maintaining the hyaluronan matrix around heart anlage Reviewed

    Yuzuru Ito, Satsuki Seno, Hiroaki Nakamura, Akimasa Fukui, Makoto Asashima

    DEVELOPMENTAL BIOLOGY   319 ( 1 )   34 - 45   2008.7

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    Hyaluronan matrix plays an important role during vertebrate cardiogenesis. Transcripts for the hyaluronan synthase Has2 gene are expressed in heart anlage, and disruption of either Has2 or versican, a hyaluronan matrix component, abrogates normal cardiac morphogenesis. However, the mechanisms by which hyaluronan matrix contributes to early heart development are largely unknown. Here we show that Xenopus hyaluronan and proteoglycan-binding link protein 3 (XHAPLN3) helps to maintain hyaluronan matrix around the cardiac anlage, and thereby contribute to cardiogenesis. XHAPLN3 mRNA transcript localization overlapped with the mRNA expression of both Xhas2 and Xversican at the heart anlage of early tailbud (stage 23) embryos. Furthermore, knockdown of XHAPLN3 or Xhas2 with morpholino antisense oligos caused a heart deficiency in developing tadpoles. Our results show when and how components of the hyaluronan matrix function in cardiogenesis, improving our understanding of how extracellular matrix participates in embryogenesis. (C) 2008 Elsevier Inc. All rights reserved.

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  • 原腸陥入運動とケモカインシグナル Reviewed

    福井彰雅

    生物物理   48 ( 1 )   23 - 29   2008.2

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  • Blood cell and vessel formation following transplantation of activin-treated explants in Xenopus Reviewed

    Kentaro Nagamine, Miho Furue, Akirnasa Fukui, Akira Matsuda, Takarmtsu Hori, Makoto Asashima

    BIOLOGICAL & PHARMACEUTICAL BULLETIN   30 ( 10 )   1856 - 1859   2007.10

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    Treatment of Xenopus blastula with activin converts undifferentiated presumptive ectoderm (animal cap) into mesoderm and endoderm in a dose-dependent manner. At low concentrations, activin induces ventral mesoderm such as blood cells. Here we show that activin-treated aggregates of animal cap cells prepared from undifferentiated presumptive ectoderm and transplanted into Xenopus embryos differentiated to form red blood cells and vascular endothelial cells. We compared gene expression profiles of the activin-treated with untreated aggregates of animal cap cells using microarray analysis. This revealed 838 clones including vascular-related genes that were expressed at levels at least 2-fold greater in the activin-treated aggregates than in the untreated controls. Of these, 356 were known Xenopus genes, 296 had homologues, and 186 were unknown genes. These findings identified novel vascular-related genes and provided insights into how the blood vessel system establishes in normal development.

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  • Hydrodynamic properties of DNA and DNA-lipid complex in an elongational flow field Reviewed

    Naoki Sasaki, Hidetomo Ashitaka, Kenji Ohtomo, Akimasa Fukui

    INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES   40 ( 4 )   327 - 335   2007.3

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    The aim of this study was to determine the difference between hydrodynamic properties of DNA-cetyltrimethylammonium (CTA) complex and those of DNA, which may be related to the difference in fibre-forming ability of DNA-CTA from that of DNA. Responses of DNA and DNA-CTA complex to an elongational flow field were investigated. In both solution systems, results suggesting a coil-stretch transition were obtained. From a critical strain rate value, the radius of gyration of DNA-CTA molecules in ethanol-glycerol solution was revealed to be 0.3-0.5 times of that of DNA in aqueous NaCl solution. Shear viscosity of DNA-CTA solution was much smaller than that of DNA solution, also suggesting a smaller size of DNA-CTA in ethanol-glycerol solution than that of DNA in aqueous NaCl solution. The plateau birefringence value of the DNA-CTA system, a parameter that indicates the local molecular conformation and the molecular arrangement, was only about 1/10 of that of the DNA system. There is an empirically determined molecular model of DNA-CTA complex in which a DNA molecule is sheathed by a cylindrical crust made of CTA chains. This structure reduces the DNA molecular density in a pure elongational flow field region but cannot explain the observed reduction of birefringence intensity. The small plateau birefringence value of DNA-CTA compared with that of DNA was attributed to the reduced molecular polarizability by the particular conformation of DNA molecules and CTA chains in the DNA-CTA system such as that expected by the conformational models. (c) 2006 Elsevier B.V. All rights reserved.

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  • A novel gene, BENI is required for the convergent extension during Xenopus laevis gastrulation Reviewed

    Motohiro Homma, Masafumi Inui, Akimasa Fukui, Tatsuo Michiue, Koji Okabayashi, Makoto Asashima

    DEVELOPMENTAL BIOLOGY   303 ( 1 )   270 - 280   2007.3

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    Activin-like signaling plays an important role in early embryogenesis. Activin A, a TGF beta family protein, induces mesodermal/endodermal tissues in animal cap assays. In a screen for genes expressed early after treatment with activin A, we isolated a novel gene, denoted as BENI (Brachyury Expression Nuclear Inhibitor). The BENI protein has a conserved domain at the N-terminus that contains a nuclear localization signal (NLS), and two other NLSs in the C-terminal domain. BENImRNA was localized to the animal hemisphere at the gastrula stages and to ectoderm except for neural regions at stage 17; expression persisted until the tadpole stage. The overexpression of BENI caused gastrulation defects and inhibition of elongation of activin-treated animal caps with reduction of Xbra expression. Moreover, whole-mount in situ hybridization revealed reduced expression of Xbra in BENI mRNA-injected regions of gastrula embryos. Functional knockdown of BENI using an antisense morpholino oligonucleotide also resulted in an abnormal phenotype of embryos curling to the dorsal side, and excessive elongation of activin-treated animal caps without altered expression of mesodermal markers. These results suggested that BENI expression is regulated by activin-like signaling, and that this regulation is crucial for Xbra expression. (c) 2006 Elsevier Inc. All rights reserved.

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  • Xapelin and Xmsr are required for cardiovascular development in Xenopus laevis Reviewed

    Masafumi Inui, Akimasa Fukui, Yuzuru Ito, Makoto Asashima

    DEVELOPMENTAL BIOLOGY   298 ( 1 )   188 - 200   2006.10

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    The cardiovascular development is the elaborate process, and despite the extensive studies, the mechanisms underlying endothelial, hematopoietic, and cardiac developments, as well as the interrelation between these processes, are not fully understood. In this study, we demonstrated that Xenopus apelin and Xmsr play pivotal roles in cardiovascular development. Apelin is a recently identified ligand for an orphan G-protein-coupled receptor APJ and is involved in fluid homeostasis in mammals. Xenopus preproapelin (Xpreproapelin) was isolated and its mRNA localized to the region around the presumptive blood vessels, which are overlapping or adjacent to those expressing Xmsr, the Xenopus homologue of APJ Overexpression of Xpreproapelin disorganized the expression of the endothelial precursor cell marker XlFli and the hematopoietic precursor cell marker SCL at the neurula, whereas embryos injected with morpholino antisense oligonucleotides for Xapelin and Xmsr displayed attenuated expression of Tie2, alpha-globin, XPOX2, and cTnI, markers of endothelium, erythrocytes, myeloid cells, and cardiomyocytes, respectively. XlFli morpholino had similar effects to Xapelin and Xmsr morpholinos on cardiac differentiation, suggesting an unexpected potential relationship between the endothelium and cardiac differentiation. Forced expression of constitutive active G alpha i rescued the phenotypes of Xmsr morpholino-injected embryos, indicating that the i/o type of G protein alpha subunit acts downstream of Xmsr. (c) 2006 Elsevier Inc. All rights reserved.

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  • Novel Daple-like protein positively regulates both the Wnt/beta-catenin pathway and the Wnt/JNK pathway in Xenopus Reviewed

    H Kobayashi, T Michiue, A Yukita, H Danno, K Sakurai, A Fukui, A Kikuchi, M Asashima

    MECHANISMS OF DEVELOPMENT   122 ( 10 )   1138 - 1153   2005.10

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    Wnt signaling pathways are essential in various developmental processes including differentiation, proliferation, cell migration, and cell polarity. Wnt proteins execute their multiple functions by activating distinct intracellular signaling cascades, although the mechanisms underlying this activation are not fully understood. We identified a novel Daple-like protein in Xenopus and named it xDal (Xenopus Daple-like). As with Daple, xDal contains several leucine zipper-like regions (LZLs) and a putative PDZ domain-binding motif, and can interact directly with the dishevelled protein. In contrast to mDaple, injection of xDal mRNA into the dorso-vegetal blastomere does not induce ventralization and acted synergistically with xdsh in secondary axis induction. XDal also induced expression of siamois and xnr-3, suggesting that XDal functions as a positive regulator of the Wnt/-catenin pathway. Injection of xDal mRNA into the dorso-animal blastomere, however, induced gastrulation-defective phenotypes in a dose-dependent manner. In addition, xDal inhibited activin-induced elongation of animal caps and enhanced c-jun phosphorylation. Based on these findings, xDal is also thought to function in the Wnt/JNK pathway. Moreover, functional domain analysis with several deletion mutants indicated that xDaI requires both a putative PDZ domain-binding motif and at least one LZL for its activity. These findings with xDal will provide new information on the Wnt signaling pathways. 2005 Elsevier Ireland Ltd. All rights reserved.

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  • Transcriptional profiles of activin-treated explants in Xenopus laevis Reviewed

    A. Fukui, M. Asashima

    MECHANISMS OF DEVELOPMENT   122   S140 - S140   2005.9

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  • Induction of cells expressing vascular endothelium markers from undifferentiated Xenopus presumptive ectoderm by co-treatment with activin and angiopoietin-2 Reviewed

    K Nagamine, M Furue, A Fukui, M Asashima

    ZOOLOGICAL SCIENCE   22 ( 7 )   755 - 761   2005.7

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    Activin is a potent inducer of mesoderm in amphibian embryos. We previously reported that low concentrations of activin could induce the formation of blood cells from Xenopus explants (animal caps). Both hematopoietic and vascular endothelial cell lineages are believed to share a common precursor, termed hemangioblasts. In this study, we tried to induce differentiation of vascular endothelial cells in aggregates derived from Xenopus animal caps. Aggregates formed from cells that were co-treated with activin and angiopoietin-2 expressed the vascular endothelial markers, X-msr, Xtie2 and Xegf17. However, none of these aggregates expressed the hematopoietic marker genes, globin alpha T3, alpha T5, alpha A or GATA-1. We used microarray analysis to compare the gene expression profiles of aggregates treated with activin alone or with activin and angiopoietin. The combination, but not activin alone, induced expression of vascular-related genes such as XI-fli and VEGF. These results demonstrate that treatment of dissociated animal cap cells with activin and angiopoietin-2 can induce differentiation of endothelial cells, and provides a promising model system for the in vitro study of blood vessel induction in vertebrates.

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  • Temporal regulation of global gene expression and cellular morphology in Xenopus kidney cells in response to clinorotation Reviewed

    J Kitamoto, A Fukui, M Asashima

    SPACE LIFE SCIENCES: CLOSED ECOLOGICAL SYSTEMS: EARTH AND SPACE APPLICATIONS   35 ( 9 )   1654 - 1661   2005

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    Here, we report changes gene expression and morphology of the renal epithelial cell line, A6, which was derived from Xenopus laevis adult kidney that had been induced by long-term culturing with a three-dimensional clinostat. An oligo microarray analysis on the A6 cells showed that mRNA levels for 52 out of 8091 genes were significantly altered in response to clinorotation. On day 5, there was no dramatic change in expression level, but by day 8 and day 10, either upregulation or downregulation of gene expression became evident. By day 15, the expression levels of 18 out of 52 genes had returned to the original levels, while the remaining 34 genes maintained the altered levels of expression. Quantitative analyses of gene expression by real-time PCR confirmed that changes in the mRNA levels of selected genes were found only under clinorotation and not under hypergravity (7g) or ground control. Morphological changes including loss of dome-like structures and disorganization of both E-cadherin adherence junctions and cortical actin were also observed after 10 days of culturing with clinorotation. These results revealed that the expression of selected genes was altered specifically in A6 cells cultured under clinorotation. (c) 2005 COSPAR. Published by Elsevier Ltd. All rights reserved.

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  • Global and temporal regulation of gene expression in Xenopus kidney cells in response to presumed microgravity generated by 3D clinostats.

    Junko Kitamoto, Akimasa Fukui, Makoto Asashima

    Uchu Seibutsu Kagaku   18 ( 3 )   152 - 3   2004.11

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    We documented changes in morphology and gene expression of the renal epithelial cell line A6 derived from Xenopus leavis adult kidney induced by long-term culturing with three dimensional clinostats. An oligo microarray analysis on A6 cells showed that mRNA levels of 52 out of 8091 genes were significantly altered in response to clinorotation. Upregulation or downregulation of gene expression became evident on day 8 and day 10 while there was no significant change on day 5. However, on day 15, expression of 18 out of 52 genes resumed to the levels similar to its original levels while remaining 33 genes maintained altered levels of expression. Quantitative analyses of gene expression by real-time PCR confirmed that changes in mRNA levels of selected genes were found only under clinorotation but not under hypergravity (7 G) and ground control (1 G). Morphological changes including loss of dome-like structures, disassembly of E-cadherin adherence junctions and disassembly of cortical actin were also observed over 10 days of culturing with clinorotation. The results revealed genes which expression was altered specifically in A6 cells cultured under clinorotation.

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  • Profiles of gene expression by activin-A in Xenopus presumptive ectoderm Reviewed

    Akimasa Fukui, Motohiro Homma, Makoto Asashima

    10th International Xenopus Meeting Program Book   64   2004.9

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  • XSENP1, a novel sumo-specific protease in Xenopus, inhibits normal head formation by down-regulation of Wnt/beta-catenin signalling Reviewed

    A Yukita, T Michiue, A Fukui, K Sakurai, H Yamamoto, M Ihara, A Kikuchi, M Asashima

    GENES TO CELLS   9 ( 8 )   723 - 736   2004.8

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    Small Ubiqutin-related modifier (SUMO), which is responsible for the ubiquitination-like post-translational modification 'sumoylation', regulates a number of biological processes including, in particular, transcription. The rat protein Axam, which possesses SUMO-specific protease activity, was shown to inhibit the Wnt signalling pathway. Several other components of the pathway are also sumoylated, so the mechanism of this modification has itself been linked to Wnt signalling. However, the functional interactions between SUMO and Wnt signalling are not well understood. This study identified a novel SUMO-specific protease in Xenopus, which was denoted XSENP1. The C-terminus of XSENP1 is highly conserved across the SUMO-specific protease family, and in vitro XSENP1 possesses hydrolase and desumoylation activity. Over-expression of XSENP1 in vivo inhibited dorso-anterior development of Xenopus embryos and suppressed Wnt signalling target gene expression in a manner similar to Axam. Deletion analysis of XSENP1 showed that inhibition of the Wnt signalling pathway requires protease activity. Moreover, XSENP1 inhibits ectopic axis induction by Dvl, beta-catenin and the constitutively active form of beta-catenin, but not by siamois. These results indicate that the dorsal expression of XSENP1 obstructs head development in Xenopus laevis and that this effect may result from inhibition of the canonical Wnt pathway downstream of beta-catenin, but upstream of siamois.

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  • Xldax, an inhibitor of the canonical Wnt pathway, is required for anterior neural structure formation in Xenopus Reviewed

    T Michiue, A Fukui, A Yukita, K Sakurai, H Danno, A Kikuchi, M Asashima

    DEVELOPMENTAL DYNAMICS   230 ( 1 )   79 - 90   2004.5

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    Wnt signaling pathways are involved during various stages in the development of many species. In Xenopus, the accumulation of beta-catenin on the dorsal side of embryo is required for induction of the organizer, while the head structure formation requires inhibition of Wnt signaling. Here, we report a role for xldax, a negative regulator of Wnt signaling. Xldax is expressed in neural tissues at the neurula stage, and in the restricted region of the tadpole brain. Ectopic expression of xldax inhibits the target gene expression, suggesting that xldax can inhibit canonical Wnt signaling. To examine the function of xldax, a morpholino oligo for xldax (xldaxMO) was designed. An injection into an animal pole cell caused a loss of forebrain. The anterior neural marker expression is decreased in xldaxMO-injected embryo, suggesting that xldax is required for anterior neural development. Moreover, a negative regulator that acts downstream of xldax rescued this defect. We propose that Idax functions are dependent on the canonical Wnt pathway and are crucial for the anterior neural development. (C) 2004 Wiley-Liss, Inc.

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  • Duplin, a novel Wnt signaling regulation factor, inhibit axis formation in Xenopus embryos

    Akimasa Fukui

    The 6th conference Asia-Pacific International Molecular Biology Network Abstracts   35   2003.11

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  • Characterization of a gene respondent to clinorotation in Xenopus A6 cells. Reviewed

    Kyuno J, Fukui A, Michiue T, Asashima M

    Uchu Seibutsu Kagaku   17 ( 3 )   171 - 172   2003.10

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  • Identification and characterization of Xenopus NDRG1 Reviewed

    J Kyuno, A Fukui, T Michiue, M Asashima

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   309 ( 1 )   52 - 57   2003.9

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    NDRG1 is a member of the N-myc downstream-regulated gene (NDRG) family and is involved in cellular differentiation, activation of p53, cell cycle arrest, metastasis, and hypoxia. Expression of NDRG1 is repressed by the proto-oncogene, N-myc during mouse development, although the exact functional role of NDRG1 in development remains unknown. Here, we report the characterization of Xenopus laevis NDRG1 (xNDRG1) during X laevis development. Expression of xNDRG1 transcript was first detected at stage 15, and was localized to the presumptive pronephric anlagen at stage 26 and to pronephros, eye, branchial arches, and tail-bud at stage 32. Overexpression of xNDRG1 results in a reduced pronephros and disorganized somites. Depletion of xNDRG1, using morpholinos, causes failure of pronephros development. These results suggest that xNDRG1 is required for pronephros development in X. laevis. (C) 2003 Elsevier Inc. All rights reserved.

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  • Immunocytochemical study of activin type IB receptor (XALK4) in Xenopus oocytes Reviewed

    A Fukui, S Komazaki, O Miyoshi, M Asashima

    DEVELOPMENT GROWTH & DIFFERENTIATION   45 ( 2 )   113 - 119   2003.4

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    Studies have shown that the activin type IB receptor is specific for activin/nodal signaling. Activin is produced by follicle cells in the ovary, and is incorporated into the oocytes. Antisera against three peptides were prepared, encompassing the extracellular, intracellular and serine/threonine kinase domains of the Xenopus type IB activin receptor (XALK4). Immunocytochemistry was done using these antisera to investigate the distribution of XALK4 in the Xenopus ovary. All three antisera stained the mitochondrial cloud of Xenopus previtellogenic oocytes. Purified antibody against the intracellular domain also recognized the mitochondrial cloud. Immunoelectron microscopy localized XALK4 on the endoplasmic reticulum of the mitochondrial cloud, although not on mitochondria.

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  • Desumoylation activity of Axam, a novel Axin-binding protein, is involved in downregulation of beta-catenin Reviewed

    T Kadoya, H Yamamoto, T Suzuki, A Yukita, A Fukui, T Michiue, T Asahara, K Tanaka, M Asashima, A Kikuchi

    MOLECULAR AND CELLULAR BIOLOGY   22 ( 11 )   3803 - 3819   2002.6

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    Axam has been identified as I novel Axin-binding protein that inhibits the Wnt signaling pathway. We studied the molecular mechanism by which Axam stimulates the downregulation of beta-catenin. The C-terminal region of Axam has an amino acid sequence similar to that of the catalytic region of SENP1, a SUMO-specific protease (desumoylation enzyme), Indeed, Axam exhibited activity to remove SUMO from sumoylated proteins in vitro and in intact cells. The Axin-binding domain is located in the central region of Axam, which is different from the catalytic domain. Neither the Axin-binding domain nor the catalytic domain alone was sufficient for the downregulation of beta-catenin. An Axam fragment which contains both domains was able to decrease the level of beta-catenin. On substitution of Ser for Cys(547) in the catalytic domain, Axam lost its desumoylation activity. Further, this Axam mutant decreased the activity to downregulate beta-catenin. Although Axam strongly inhibited axis formation and expression of siamois, a Wnt-response gene, in Xenopus embryos, Axam(C547S) showed weak activities. These results demonstrate that Axam functions as a desumoylation enzyme to downregulate beta-catenin and suggest that sumoylation is involved in the regulation of the Wnt signaling pathway.

    DOI: 10.1128/MCB.22.11.3803-3819.2002

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  • Synthesis and release of activin and noggin by cultured human amniotic epithelial cells Reviewed

    S Koyano, A Fukui, S Uchida, K Yamada, M Asashima, N Sakuragawa

    DEVELOPMENT GROWTH & DIFFERENTIATION   44 ( 2 )   103 - 112   2002.4

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    Recent studies suggest that extra-embryonic tissues may be essential sources of early organizing signals for the mouse embryo. In vitro studies of human amniotic epithelial cells (HAEC) have shown that the amnion can produce various biologically active substances. In this study, the synthesis and release of activin A and noggin, and the activin signaling pathway, was investigated in HAEC. Conditioned medium from cultured HAEC contained activin A which was functionally active in Xenopus laevis animal cap assays. Immunohistochemistry, western blotting and reverse transcription-polymerase chain reaction confirmed that HAEC also synthesize and release noggin. Noggin transcripts were induced by the addition of recombinant activin A, and activin A was inhibited by activin antibody except in the presence of cycloheximide (CHX). These data demonstrate that noggin mRNA expression is induced directly by activin A without new protein synthesis, indicating that noggin is a primary response gene. The results suggest that there is an activin signaling pathway in HAEC, and that the human amnion might therefore be involved in neural formation during early development.

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  • Nuclear localization of Duplin, a beta-catenin-binding protein, is essential for its inhibitory activity on the Wnt signaling pathway Reviewed

    M Kobayashi, S Kishida, A Fukui, T Michiue, Y Miyamoto, T Okamoto, Y Yoneda, M Asashima, A Kikuchi

    JOURNAL OF BIOLOGICAL CHEMISTRY   277 ( 8 )   5816 - 5822   2002.2

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    Duplin binds to beta-catenin and inhibits the Wnt signaling pathway, thereby leading to repression of the beta-catenin-mediated transactivation and Xenopus axis formation. To find an additional function of Duplin, yeast two-hybrid screening was carried out. Importin a was isolated as a binding protein of Duplin. Importin a bound directly to basic amino acid clusters of Duplin. Although Duplin was present in the nucleus, deletion of the basic amino acid clusters (Duplin(Delta500-584)) retained Duplin in the cytoplasm. Duplin(Delta500-584) bound to beta-catenin as efficiently as wild-type Duplin, but it neither repressed Wnt-dependent Tcf transcriptional activation in mammalian cells nor showed ventralization in Xenopus embryos. The Duplin mutant without a beta-catenin-binding region lost the ability to inhibit the Wnt-dependent Tcf activation, but retained its ventralizing activity. Furthermore, Duplin not only suppressed beta-catenin-dependent axis duplication and expression of siamois, a Wnt-regulated gene, but also inhibited siamois-dependent axis duplication. These results indicate that Duplin is translocated to the nucleus by interacting with importin a, and that nuclear localization is essential for the function of Duplin. Moreover, Duplin has an additional activity of inhibiting the Wnt signaling pathway by affecting the downstream beta-catenin target genes.

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  • Role of BMP-4 in the inducing ability of the head organizer in Xenopus laevis Reviewed

    A Sedohara, A Fukui, T Michiue, M Asashima

    ZOOLOGICAL SCIENCE   19 ( 1 )   67 - 80   2002.1

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    BMP-4 has been implicated in the patterning of the Dorsal-Ventral axis of mesoderm and ectoderm. In this study, we describe the posteriorizing effect of BMP-4 on the neural inducing ability of dorsal mesoderm (dorsal lip region) in Xenopus gastrulae. Dorsal lip explants dissected from stage 10.25 embryos retained anterior inducing ability when precultured for 6 hrs until sibling embryos reach stage 12. When the dorsal lips from stage 10.25 embryos were treated with a range of BMP-4 concentrations, posterior tissues were induced in adjacent ectoderm in a dose-dependent manner. Thus activin-treated explants able to act as head inducers can also induce posterior structures in the presence of BMP-4. To investigate whether BMP-4 directly affects the inducing ability of dorsal mesoderm, we blocked the BMP4 signaling pathway by injection of mRNA encoding a truncated form of the BMP-4 receptor (tBR) mRNA. Under these conditions, activin-treated explants induced anterior tissues following BMP-4 treatment. Taken together, these results indicate that BMP-4 may affect the head inducing ability of dorsal mesoderm and confer trunk-tail inducing ability during Xenopus gastrulation.

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  • Synergistic activation of the Wnt signaling pathway by Dvl and casein kinase I epsilon Reviewed

    M Kishida, S Hino, T Michiue, H Yamamoto, S Kishida, A Fukui, M Asashima, A Kikuchi

    JOURNAL OF BIOLOGICAL CHEMISTRY   276 ( 35 )   33147 - 33155   2001.8

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    Although casein kinase I epsilon (CKI epsilon) has been shown to regulate the Wnt signaling pathway positively, its mode of action is not clear. In this study we show that CKI epsilon activates the Wnt signaling pathway in co-operation with Dv1. CKI epsilon and Axin associated with different sites of Dv1, and CKI epsilon and Dv1 interacted with distinct regions on Axin. Therefore, these three proteins formed a ternary complex. Either low expression of Dv1 or CKI epsilon alone did not accumulate beta -catenin, but their co-expression accumulated greatly. Dv1 and CKI epsilon activated the transcriptional activity of T cell factor (Tcf) synergistically. Although the Dv1 mutant that binds to Axin but not to CKI epsilon activated Tcf, it did not synergize with CKI epsilon Another Dv1 mutant that does not bind to Axin did not activate Tcf irrespective of the presence of CKI epsilon Furthermore, Dv1 and CKI epsilon co-operatively induced axis duplication of Xenopus embryos. These results indicate that Dv1 and CKI epsilon synergistically activated the Writ signaling pathway and that the binding of the complex of Dv1 and CKI epsilon to Axin is necessary for their synergistic action.

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  • Inhibition of the Wnt signaling pathway by the PR61 subunit of protein phosphatase 2A Reviewed

    H Yamamoto, T Hinoi, T Michiue, A Fukui, H Usui, Janssens, V, C Van Hoof, J Goris, M Asashima, A Kikuchi

    JOURNAL OF BIOLOGICAL CHEMISTRY   276 ( 29 )   26875 - 26882   2001.7

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    Axin, a negative regulator of the Wnt signaling pathway, forms a complex with glycogen synthase kinase-3 beta (GSK-3 beta), beta -catenin, adenomatous polyposis coli (APC) gene product, and Dvl, and it regulates GSK-3 beta -dependent phosphorylation in the complex and the stability of beta -catenin. Using yeast two-hybrid screening, we found that regulatory subunits of protein phosphatase 2A, PR61 beta and -gamma, interact with Axin. PR61 beta or -gamma formed a complex with Axin in intact cells, and their interaction was direct. The binding site of PR61 beta on Axin was different from those of GSK-3 beta, beta -catenin, APC, and Dvl. Although PR61 beta did not affect the stability of beta -catenin, it inhibited Dvl- and beta -catenin-dependent T cell factor activation in mammalian cells. Moreover, it suppressed beta -catenin-induced axis formation and expression of siamois, a Wnt target gene, in Xenopus embryos, suggesting that PR61 beta acts either at the level of beta -catenin or downstream of it. Taken together with the previous observations that PR61 interacts with APC and functions upstream of beta -catenin, these results demonstrate that PR61 regulates the Wnt signaling pathway at various steps.

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  • Inhibition of the Wnt signaling pathway by Idax, a novel Dvl-binding protein Reviewed

    SI Hino, S Kishida, T Michiue, A Fukui, Sakamoto, I, S Takada, M Asashima, A Kikuchi

    MOLECULAR AND CELLULAR BIOLOGY   21 ( 1 )   330 - 342   2001.1

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    In attempting to clarify the roles of Dvl in the Wnt signaling pathway, we identified a novel protein which binds to the PDZ domain of Dvl and named it Idax (for inhibition of the Dvl and Axin complex). Idax and Axin competed with each other for the binding to Dvl. Immunocytochemical analyses showed that Idax was localized to the same place as DvI in cells and that expression of Axin inhibited the colocalization of Dvl and Idax. Further, Wnt-induced accumulation of beta -catenin and activation of T-cell factor in mammalian cells were suppressed by expression of Idax. Expression of Idax in Xenopus embryos induced ventralization with a reduction in the expression of siamois, a Wnt-inducible gene. Idax inhibited Wnt- and Dvl- but not beta -catenin-induced axis duplication. It is known that Dvl is a positive regulator in the Wnt signaling pathway and that the PDZ domain is important for this activity. Therefore, these results suggest that Idax functions as a negative regulator of the Wnt signaling pathway by directly binding to the PDZ domain of Dvl.

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  • Meiotic maturation induces animal-vegetal asymmetric distribution of aPKC and ASIP/PAR-3 in Xenopus oocytes Reviewed

    M Nakaya, A Fukui, Y Izumi, K Akimoto, M Asashima, S Ohno

    DEVELOPMENT   127 ( 23 )   5021 - 5031   2000.12

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    The asymmetric distribution of cellular components is an important clue for understanding cell fate decision during embryonic patterning and cell functioning after differentiation. In C, elegans embryos, PAR-3 and aPKC form a complex that colocalizes to the anterior periphery of the one-cell embryo, and are indispensable for anterior-posterior polarity that is formed prior to asymmetric cell division. In mammals, ASIP (PAR-3 homologue) and aPKC lambda form a complex and colocalize to the epithelial tight junctions, which play critical roles in epithelial cell polarity. Although the mechanism by which PAR-3/ASIP and aPKC regulate cell polarization remains to be clarified, evolutionary conservation of the PAR-3/ASIP-aPKC complex suggests their general role in cell polarity organization. Here, we show the presence of the protein complex in Xenopus laevis, In epithelial cells, XASIP and XaPKC colocalize to the cell-cell contact region. To our surprise, they also colocalize to the animal hemisphere of mature oocytes, whereas they localize uniformly in immature oocytes. Moreover, hormonal stimulation of immature oocytes results in a change in the distribution of XaPKC 2-3 hours after the completion of germinal vesicle breakdown, which requires the kinase activity of aPKC, These results suggest that meiotic maturation induces the animal-vegetal asymmetry of aPKC.

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  • Inhibition of Wnt signaling pathway by a novel axin-binding protein Reviewed

    T Kadoya, S Kishida, A Fukui, T Hinoi, T Michiue, M Asashima, A Kikuchi

    JOURNAL OF BIOLOGICAL CHEMISTRY   275 ( 47 )   37030 - 37037   2000.11

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    Axin forms a complex with adenomatous polyposis coli gene product, glycogen synthase kinase-3 beta (GSK-3 beta), beta -catenin, DvI, and protein phosphatase 2A and functions as a scaffold protein in the Wnt signaling pathway. In the Axin complex, GSK-3 beta efficiently phosphorylates beta -catenin, which is then ubiquitinated and degraded by proteasome. We isolated a novel protein that binds to Axin and named it Axam (for Axin associating molecule). Axam formed a complex with Axin in intact cells and bound directly to Axin. Axam inhibited the complex formation of DvI with Axin and the activity of DvI to suppress GSK-3 beta -dependent phosphorylation of Axin. Furthermore, Axam induced the degradation of beta -catenin in SW480 cells and inhibited Wnt-dependent axis duplication in Xenopus embryos. These results suggest that Axam regulates the Wnt signaling pathway negatively by inhibiting the binding of DvI to Axin.

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  • Effects of rat Axin domains on axis formation in Xenopus embryos Reviewed

    A Fukui, S Kishida, A Kikuchi, M Asashima

    DEVELOPMENT GROWTH & DIFFERENTIATION   42 ( 5 )   489 - 498   2000.10

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    Wnt signaling plays an important role in axis formation in early vertebrate development. Axin is one Wnt signaling regulator that inhibits this pathway. The effects of the injection of mRNA of several rat Axin (rAxin) mutants on axis formation in Xenopus embryos were examined. It was found that rAxin mutants containing only a regulation of G-protein signaling (RGS) domain fragment or with deletion of the RGS domain induced axis formation. Because the RGS domain is a major adenomatous polyposis coli gene product (APC)-binding domain, APC association with glycogen synthase kinase 3 beta (GSK3 beta) on the Axin molecule may be important in inhibition of axis formation. The ventralizing activities of wild-type rAxin and a mutant in which the Dishevelled and Axin (DIX) domain was deleted (Delta DIX mutant) were examined. Histological examination and gene expression revealed that the ventralizing activity of the Delta DIX mutant was weaker than that of wild-type rAxin. This finding suggests that the C-terminus of rAxin contributes to the inhibition of Wnt signaling in Xenopus embryos. Furthermore, an rAxin mutant that contained both the RGS and GSK3 beta -binding domains affected both the dorsal and ventral sides of blastomeres, mediated ectodermal fate and induced expansion of notochord and/or endoderm, but did not induce axis formation.

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  • A novel beta-catenin-binding protein inhibits beta-catenin-dependent Tcf activation and axis formation Reviewed

    Sakamoto, I, S Kishida, A Fukui, M Kishida, H Yamamoto, S Hino, T Michiue, S Takada, M Asashima, A Kikuchi

    JOURNAL OF BIOLOGICAL CHEMISTRY   275 ( 42 )   32871 - 32878   2000.10

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    beta -Catenin is efficiently phosphorylated by glycogen synthase kinase-3 beta in the Axin complex in the cytoplasm, resulting in the down-regulation. In response to Wnt, beta -catenin is stabilized and translocated into the nucleus where it stimulates gene expression through Tcf/Lef, Here we report a novel protein, designated Duplin (for axis duplication inhibitor), which negatively regulates the function of beta -catenin in the nucleus. Duplin was located in the nucleus. Duplin bound directly to the Armadillo repeats of beta -catenin, thereby inhibiting the binding of Tcf to beta -catenin. It did not affect the stability of beta -catenin but inhibited Wnt- or beta -catenin-dependent Tcf activation. Furthermore, expression of Duplin in Xenopus embryos inhibited the axis formation and beta -catenin-dependent axis duplication, and prevented the beta -catenin's ability to rescue ventralizing phenotypes induced by ultraviolet light irradiation. Thus, Duplin is a nuclear protein that inhibits beta -catenin signaling.

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  • Spontaneous thyroid-containing teratoma associated with impaired development in the African clawed frog, Xenopus laevis Reviewed

    SW Cheong, A Fukui, M Asashima, CJ Pfeiffer

    JOURNAL OF COMPARATIVE PATHOLOGY   123 ( 2-3 )   110 - 118   2000.8

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    Teratomas are rare in amphibians and the neoplasm described here, which had a significant thyroid carcinoma component, is the first tumour of this type to be reported in Xenopus laevis. The thyroid component contained moderately to well-differentiated acinar glands showing much hyperplasia, dysplasia, and reduced and distorted colloid reservoirs. Cartilaginous, neural, muscular, mesenchymal and gut-like epithelial components were also observed in this ventral mediastinal neoplasm, indicating aberrant proliferation from all three germ layers. This teratoma was only one abnormality in a complex of developmental changes, followed for 28 months, which appeared in a single generation of sibling 2-week-old Xenopus larvae. Two hundred larvae produced by an apparently normal adult pair initially showed ocular defects. including microphthalmia, anophthalmia and tumours projecting near the eyes. During further development up to 28 months, mediastinal tumours developed in nine frogs; these tumours were associated with reduced growth, the frogs reaching only 13-20% of normal weight, and greater enhanced ventral pigmentation. (C) 2000 Harcourt Publishers Ltd.

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  • Expression of the Xenopus GTP-binding protein gene Ran during embryogenesis Reviewed

    Y Onuma, R Nishihara, S Takahashi, K Tanegashima, A Fukui, M Asashima

    DEVELOPMENT GENES AND EVOLUTION   210 ( 6 )   325 - 327   2000.6

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    The Ran gene family encodes small GTP binding proteins that are associated with a variety of nuclear processes. We isolated a Xenopus Ran cDNA and analyzed the pattern of expression of this gene during embryogenesis. Ran is expressed maternally and later in the CNS, neural crest, mesenchyme, eyes, and otic vesicles. However, expression is not detected in the somites or the notochord.

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  • XSIP1, a member of two-handed zinc finger proteins, induced anterior neural markers in Xenopus laevis animal cap Reviewed

    A Eisaki, H Kuroda, A Fukui, M Asashima

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   271 ( 1 )   151 - 157   2000.4

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    We characterized Xenopus SIP1 (XSIP1), Smad interacting protein, from activin-treated animal caps by differential screening. The XSIP1 is very similar to mouse SIP1 in the protein coding region including the zinc huger domain and homeodomain. The expression pattern was analyzed by RT-PCR and whole mount in situ hybridization. XSIP1 expression was initially restricted to the dorsal marginal zone in the late gastrula and was subsequently expressed at the lateral edge of neural plate and, in the tailbud stage, in the forebrain, neural tube, and eye. Overexpression of XSIP1 at the animal caps resulted in activation of anterior neural markers without mesodermal markers. Ectopic expression of XSIP1 induced enlargement of neural cells and disordered eye formation. In addition to abnormal head phenotypes, many embryos were short-tailed. Our findings suggest that XSIP1 is a transcriptional repressor, which may be involved in the activin-dependent signal pathway, (C) 2000 Academic Press.

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  • Activin incorporation into vitellogenic oocytes of Xenopus laevis Reviewed

    A Fukui, R Shiurba, M Asashima

    CELLULAR AND MOLECULAR BIOLOGY   45 ( 5 )   545 - 554   1999.7

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    Activin uptake into Xenopus oocytes was studied by several complementary methods. Immunocytochemistry of adult ovary localized activin and follistatin in the cytoplasm of vitellogenic oocytes and surrounding follicle cells. Surface plasmon resonance analysis of protein interaction kinetics indicated that while follistatin or a complex of activin-follistatin bound to yolk vitellogenin, activin alone did not. Radioactive tracer analysis measured specific incorporation of activin by viable oocytes in vitro, Together, the results suggest that vitellogenic oocytes can import activins from follicle cells and that follistatin may act as a chaperone for binding activin to vitellogenin in yolk platelets.

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  • Evidence that far-infrared radiation promotes growth of xenopus laevis

    Robert Shiurba, Tatsuo Hirabayashi, Shin Kiyokawa, Akimasa Fukui, Yuko Miyanaga, Issey Kojima, Makoto Asashima

    Advances in Space Research   23 ( 12 )   2041 - 2044   1999

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    In most ectotherms, environmental temperature has differential effects on growth and differentiation. For example, amphibian size at maturity decreases with increasing temperature. To address how radiant heat in the form of far-infrared radiation (FIR) may affect development of the aquatic ectotherm Xenopus laevis, we continuously irradiated swimming larvae as they developed into young adults. Here we report evidence that FIR promotes growth of these organisms in an aqueous environment. ©1999 COSPAR. Published by Elsevier Science Ltd.

    DOI: 10.1016/S0273-1177(99)00347-6

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  • An interferon regulatory factor-related gene (xIRF-6) is expressed in the posterior mesoderm during the early development of Xenopus laevis Reviewed

    S Hatada, M Kinoshita, S Takahashi, R Nishihara, H Sakumoto, A Fukui, M Noda, M Asashima

    GENE   203 ( 2 )   183 - 188   1997.12

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    Out of a Xenopus neurula cDNA library, we isolated a clone which encodes a 52.4-kDa protein highly similar to the mouse interferon regulatory factor, IRF-6, whose function is unknown. The mRNA of this gene, named xIRF-6, seems to be maternally transmitted, but its amount rapidly decreases after the tailbud stage. Whole-mount in situ hybridization showed that xIRF-6 mRNA is expressed in the presumptive semitic mesoderm in the late gastrula, and then confined to a segment of posterior somite during the neurula through the tailbud stage. The temporally and spatially limited expression of the xIRF-6 gene product may contribute to the transcriptional regulation of specific genes which are necessary for the development of the posterior somites. (C) 1997 Elsevier Science B.V.

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  • The Na+, K+-ATPase alpha subunit requires gastrulation in the Xenopus embryo Reviewed

    T Uochi, S Takahashi, H Ninomiya, A Fukui, M Asashima

    DEVELOPMENT GROWTH & DIFFERENTIATION   39 ( 5 )   571 - 580   1997.10

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    Na+, K+-ATPase participates in reabsorption of ions and water and produces an electrochemical gradient between the intra-and extracellular spaces across the cell membrane. It also plays an important role in many developmental phenomena such as a blastocoele formation and neural formation. To elucidate the expression pattern of Na+, K+-ATPase in the Xenopus embryo, the spatial expression patterns of the Na+, K+-ATPase a subunit were studied in a normal embryo by whole-mount in situ hybridization. These transcripts were localized around the dorsal blastopore at the gastrula stage, in the neural tube at the neurula stage, and then in the pronephros and cloaca at the tail-bud stage. To study the function of Na+, K+-ATPase in embryogenesis after mid-blastula transition, the expression of the Nat, K+-ATPase a subunit was inhibited by the injection of specific antisense RNA. Embryos injected with Na+, K+-ATPase antisense RNA showed inhibition of gastrulation. When antisense RNA was injected into the dorsal blastomeres, head differentiation was markedly inhibited. These results suggest that this transcript plays an important role during gastrulation and head differentiation.

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  • An essay on the similarities and differences between inductive interactions in anuran and urodele embryos Reviewed

    GM Malacinski, T Bessho, C Yokota, A Fukui, M Asashima

    CELLULAR AND MOLECULAR LIFE SCIENCES   53 ( 4 )   410 - 417   1997.4

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    As a first step towards providing a conceptual approach to understanding similarities and differences in the mechanisms which guide inductive interactions among related organisms (e.g. various amphibia), a set of five principles is offered here. These principles were formulated by analyzing literature examples of classical embryological phenomena and by performing experiments with activin, a peptide growth factor which is currently suspected to play for a role in mesoderm induction. Mechanisms which account, at least in part, for the observed differences between anuran and urodele inductive processes can be derived from these principles.

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  • 1995 update of morphological abnormalities in wild populations of the Japanese newt Cynopus pyrrhogaster Boie (Urodela, Salamandridae). Reviewed

    Akimasa Fukui, Makoto Asashima, Victor Benno Meyer-Rochow

    ACTA HYDROBIOLOGICA   38   19 - 24   1997

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  • COMPARISON OF MESODERM-INDUCING ACTIVITY WITH MONOMERIC AND DIMERIC INHIBIN ALPHA-SUBUNIT AND BETA-A-SUBUNIT ON XENOPUS ECTODERM Reviewed

    H NAKANO, H UCHIYAMA, A FUKUI, H SUGINO, M ASASHIMA

    HORMONE RESEARCH   44   15 - 22   1995

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    Activin possesses mesoderm-inducing activity, erythroid-differentiating activity, and follicle-stimulating hormone-releasing activity. The chemical structures of the activin molecule are formed by a combination of two beta-subunit peptides of inhibin. Inhibin is a dimer consisting of an alpha and beta subunit. To examine the mesoderm-inducing activity of these substances, we tested several configurations including : (1) two types of alpha-subunit peptide; (2) two types of inhibin A and B dimer; (3) beta(A)-subunit peptide monomer; (4) three types of activins A, AB and B, and (5) follistatin (activin-binding protein) by the animal cap assay using Xenopus laevis ectoderm, and by the erythroid-differentiating factor (EDF) test. Activins, which are composed of dimeric inhibin beta(A)- or beta(B)-subunit peptides, had the highest mesoderm-inducing and EDF activities. The monomeric beta(A)-subunit peptide exhibited mesoderm-inducing and EDF activities that were much lower than activin A. The inhibitory effect of follistatin on mesodermal induction by the beta(A)-subunit peptide was also lower than that of activin. Both inhibins A and B had very weak mesoderm-inducing activity and no EDF activity. The two types of inhibin (a)lpha-subunit monomer had little mesoderm-inducing activity and no EDF activity. The mesoderm induction caused by activin A was not suppressed by the addition of the alpha-subunit monomer and inhibin. The mesoderm-inducing activity in relation to the chemical structures of the monomeric and/or dimeric inhibin alpha and beta(A) subunits is discussed.

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  • CONTROL OF CELL-DIFFERENTIATION AND MORPHOGENESIS IN AMPHIBIAN DEVELOPMENT Reviewed

    A FUKUI, M ASASHIMA

    INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY   38 ( 2 )   257 - 266   1994.6

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    We reviewed cell differentiation and morphogenesis by mesoderm-inducing factors during amphibian embryogenesis. Recently, two kinds of growth factors, activin and FGF, have been identified as influential candidates for natural mesoderm-inducing factor in amphibian development. These factors are present in early Xenopus embryos. In particular, activin has been shown to induce many kinds of mesodermal tissues in a dose-dependent manner. Activin-treated ectodermal sheet (animal cap) actsasan organizer causing gene expression, mesoderm formation and functional events such as secondary axis formation. Follistatin, an activin-specific binding protein, also present in the early Xenopus embryo, makes a complex with activin. Follistatin protein exerts no inducing activity of Xenopus animal cap. Endogenous follistatin may, however, play the role of an activin regulation factor. Endogenous actions of activin and FGF were studied using injection of their receptor mRNAs. Disruption of the FGF signaling pathway by its non-functional dominant negative receptors produced trunk and tail defects. In the case of activin, an embryo cannot form axial structures. Animal-half blastomeres from the late 8-cell stage Xenopus embryo respond to activin, and there are prepatterns in ventral and dorsal cells from very early stages. The timing of mesoderm induction during development and the relationship between the inducing factors and competent cells are discussed in this report. Differentiation of tissues and organized formation of organs can be understood as a system of serial inductive reactions originating from the organizer. We have attempted to construct a model of organizer formation based on the results of recent studies.

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  • ISOLATION AND CHARACTERIZATION OF XENOPUS FOLLISTATIN AND ACTIVINS Reviewed

    A FUKUI, T NAKAMURA, K SUGINO, K TAKIO, H UCHIYAMA, M ASASHIMA, H SUGINO

    DEVELOPMENTAL BIOLOGY   159 ( 1 )   131 - 139   1993.9

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  • ツメガエルにおける中胚葉誘導の分子機構の解明 Reviewed

    福井彰雅, 浅島 誠

    生物物理   33 ( 1 )   41 - 49   1993.2

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  • A NEW CELL-LINE (XTY) FROM A TUMOR OF XENOPUS-LAEVIS Reviewed

    A FUKUI, A TASHIRO, H KOYAMA, Y IWAMURA, M ASASHIMA

    EXPERIENTIA   48 ( 1 )   87 - 91   1992.1

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    A new cell line (XTY) was derived from a tumor of a female Xenopus laevis. This cell line has been proliferating in standard amphibian culture medium for more than 4 years (470 generations) and has a doubling time of 75.5 h at 25-degrees-C. The cells aggregate into large groups, and their stellate morphology and the expression of desmin demonstrated by immunocytochemistry suggest that their origin is not epithelial.

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Books

  • 細胞生物学事典

    石川 統, 黒岩 常祥, 永田和宏編( Role: Joint authorエピジェネティクス、中胚葉誘導)

    朝倉書店  2005 

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    Responsible for pages:78-80,290-292   Language:Japanese  

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MISC

  • ツメガエルケモカイン遺伝子の網羅的解析:異質四倍体化による遺伝子進化

    福井彰雅, 松波雅俊

    日本分子生物学会年会プログラム・要旨集(Web)   39th   ROMBUNNO.1LBA‐009 (WEB ONLY)   2016

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    J-GLOBAL

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  • Xenopus laevis全ゲノム解析 アフリカツメガエルの性染色体とWおよびZ特異的領域の解析

    回渕 修治, 和田 美加子, 高橋 秀治, 宇野 好宣, 松田 洋一, 近藤 真理子, 福井 彰雅, 高松 信彦, 平良 眞規, 伊藤 道彦

    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集   88回・38回   [1P0563] - [1P0563]   2015.12

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    Language:English   Publisher:(公社)日本生化学会  

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  • Studies on the Formation Mechanism and the Structure of the Anisotropic Collagen Gel Prepared by Dialysis-Induced Anisotropic Gelation (vol 13, pg 29, 2012)

    Kazuya Furusawa, Shoichi Sato, Jyun-ichi Masumoto, Yohei Hanazaki, Yasuyuki Maki, Toshiaki Dobashi, Takao Yamamoto, Akimasa Fukui, Naoki Sasaki

    BIOMACROMOLECULES   13 ( 4 )   1232 - 1232   2012.4

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    DOI: 10.1021/bm300346x

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  • Role of CXCR4/SDF-1 signaling in Xenopus gastrulation

    Akimasa Fukui, Toshiyasu Goto, Junko Kitamoto, Motohiro Homma, Makoto Asashima

    ZOOLOGICAL SCIENCE   22 ( 12 )   1465 - 1465   2005.12

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  • Modification of gene expression by activin-A in Xenopus presumptive ectoderm

    Akimasa Fukui, Motohiro Homma, Makoto Asashima

    ZOOLOGICAL SCIENCE   21 ( 12 )   1299 - 1299   2004.12

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  • 研究室・研究所めぐり(49)東京大学大学院総合文化研究科広域科学専攻生命環境系浅島研究室

    福井 彰雅, 浅島 誠

    遺伝   58 ( 1 )   15 - 18   2004.1

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    CiNii Books

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  • The role of Xenopus Idax gene in Noncanonical Wnt signaling pathway

    T Michiue, K Sakurai, H Kobayashi, A Yukita, A Fukui, A Kikuchi, M Asashima

    DEVELOPMENTAL BIOLOGY   259 ( 2 )   487 - 488   2003.7

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  • 総説:未分化細胞からの臓器形成

    福井 彰雅, 浅島 誠

    炎症・再生   22 ( 1 )   21 - 27   2002

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  • 総説:ツメガエルにおける臓器形成

    福井彰雅, 有泉高史, 浅島 誠

    実験医学   ( 19 )   159 - 166   2001

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  • 総説:ツメガエル胚を用いた試験管内での前腎管の形成とその移植

    福井彰雅, 盛屋直美, 浅島 誠

    発達腎研究会誌   ( 8 )   10 - 14   2000

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  • Gene expression and the morphogenesis of amphibian cultured cells

    ICHIGI Junji, FUKUI Akimasa, ASASHIMA Makoto

    13 ( 3 )   134 - 135   1999.9

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  • Localization of the polarity proteins, aPKC end ASIP/PAR-3, in Xenopus italicize laevis/italicize.

    M Nakaya, A Fukui, Y Izumi, M Asashima, S Ohno

    DEVELOPMENTAL BIOLOGY   210 ( 1 )   240 - 240   1999.6

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  • Control of organogenesis in vitro in amphibian development

    M Asashima, T Uochi, A Fukui, T Ariizumi

    DEVELOPMENTAL BIOLOGY   186 ( 2 )   B113 - B113   1997.6

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  • 技術ノート:実験マニュアル「7.両生類の細胞培養」

    福井彰雅, 浅島 誠

    生物の科学 遺伝   50 ( 10月 )   1996.10

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  • 両生類の細胞培養 (特大号 ツメガエルから何が学べるか--実験マニュアルから分子生物学まで) -- (実験マニュアル)

    福井 彰雅, 浅島 誠

    遺伝   50 ( 10 )   73 - 75   1996.10

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  • アクチビンによる組織分化

    福井彰雅, 浅島 誠

    Molecular Medicine   33 ( 7 )   828 - 829   1996.7

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  • 実験テキスト:基礎実験IIテキスト「動物の受精と初期発生. B 脊椎動物-アフリカツメガエルの発生」

    1996

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  • オーガナイザーの本体

    福井彰雅, 浅島 誠

    医事新報   ( 3745 )   136 - 137   1996

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  • 総説:中胚葉誘導因子としてのアクチビンの作用

    福井彰雅, 浅島 誠

    RADIOISOTOPES   43 ( 43 )   649 - 650   1994

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    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (other)   Publisher:日本アイソトープ協会  

    DOI: 10.3769/radioisotopes.43.10_649

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    Other Link: http://search.jamas.or.jp/link/ui/1995242299

  • アクチビンと誘導のメカニズム−両生類胚における誘導現象の研究

    福井彰雅, 浅島 誠

    細胞工学   ( 12 )   406 - 414   1993.6

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Presentations

  • アフリカツメガエルのcxcl12同祖遺伝子はなぜ保持されているのか

    太田速斗, 森山祐佳, 福井彰雅

    第14回日本ツメガエル研究集会  2021.6 

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  • アフリカツメガエル初期発生におけるCXCL12同祖遺伝子の解析

    森山祐佳, 森山侑輝, 福井彰雅

    第14回日本ツメガエル研究集会関東支部大会(XCIJ-MA)  2019.12 

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  • アフリカツメガエルはピカイアの夢を見るか? ー異質4倍体脊椎動物ゲノムの解析ー Invited

    福井彰雅

    第 298 回 三崎談話会  ( 神奈川県・三浦市 )   2019.10 

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  • 発生・再生生物学の新規モデル生物イベリアトゲイモリの網羅的遺伝子発現解析とデータベース整備

    松波 雅俊, 鈴木 美有紀, 原本 悦和, 福井 彰雅, 井上 武, 山口 勝司, 内山 郁夫, 森 一樹, 田代 康介, 伊藤 弓弦, 竹内 隆, 鈴木 賢一, 阿形 清和, 重信 秀治, 林 利憲

    日本動物学会第90回大会  2019.9 

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  • イベリアトゲイモリマクロ ファージマーカー遺伝子の探索

    長谷部ももこ, 森山侑輝, 福井彰雅

    日本動物学会第90回大会  2019.9 

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  • ツメガエルマクロファージマーカーの探索

    柳澤マリア, 佐藤妃奈乃, 森山侑輝, 福井彰雅

    日本動物学会第90回大会  2019.9 

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  • アフリカツメガエルcxcl12同祖遺伝子の解析

    森山祐佳, 森山侑輝, 福井彰雅

    日本動物学会第90回大会  2019.9 

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  • ツメガエル肝臓由来組織常在性マクロファージ培養の試み

    古作瑛菜, 森山侑輝, 福井彰雅

    第71回動物学会関東支部大会  2019.3 

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  • ツメガエル皮膚常在性マクロファージの同定の試み

    石毛香帆, 谷川実優, 森山侑輝, 福井彰雅

    第71回動物学会関東支部大会  2019.3 

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  • アフリカツメガエルのマクロファージマーカーの探索と同定

    栁澤マリア, 佐藤圭, 森山侑輝, 福井彰雅

    第71回動物学会関東支部大会  2019.3 

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  • アフリカツメガエル非筋ミオシンII同祖遺伝子の解析

    森山祐佳, 森山侑輝, 福井彰雅

    第71回動物学会関東支部大会  2019.3 

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  • CXCL12によるイベリアトゲイモリの予定脊索前板外植体遊走の制御

    伊藤 柾, 中山優希, 森山侑輝, 福井彰雅

    第71回動物学会関東支部大会  2019.3 

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  • イベリアトゲイモリ原腸形成における遺伝子発現の解析

    鳥海夏葉, 難波櫻子, 森山侑輝, 福井彰雅

    第71回動物学会関東支部大会  2019.3 

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  • イベリアトゲイモリ組織常在性マクロファージの同定

    長谷部ももこ, 森山侑輝, 福井彰雅

    第71回動物学会関東支部大会  2019.3 

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  • 両生類ケラチンの網羅的解析

    福井彰雅, 鈴木賢一, 松波雅俊

    第12回日本ツメガエル研究集会・第4回次世代両生類研究会 合同シンポジウム  2018.9 

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  • Transcriptome resource for Pleurodeles waltl: Next-generation model species for comparative studies in developmental biology and regeneration research with vertebrate model animals.

    Masatoshi Matsunami, Miyuki Suzuki, Yoshikazu Haramoto, Akimasa Fukui, Takeshi Inoue, Katsushi Yamaguchi, Ikuo Uchiyama, Kazuki Mori, Kosuke Tashiro, Yuzuru Ito, Takashi Takeuchi, Ken-ichi T Suzuki, Kiyokazu Agata, Shuji Shigenobu, Toshinori Hayashi

    第12回日本ツメガエル研究集会・第4回次世代両生類研究会 合同シンポジウム  2018.9 

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  • ツメガエルプロテオーム解析におけるヒト分子情報の活用と課題

    加藤 奈菜, 佐藤 圭, 渡会 敦子, 福井 彰雅, 加藤 尚志

    第12回ツメガエル研究会首都圏集会 XCIJ-MA  ( 東京 )   2018.7 

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  • ケモカインによる細胞集団運動制御 Invited

    福井彰雅

    2018年度精密工学会春季大会  2018.3 

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  • X. tropicalisのトランスクリプトーム解析:バイオインフォマティクスを始めるために Invited

    福井彰雅

    NBRPネッタイツメガエル第6回技術講習会  2018.2 

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  • イベリアトゲイモリトランスクリプトーム解析: P. waltl特異的遺伝子の探索

    福井彰雅

    第二回イベリアトゲイモリ研究会  2017.12 

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  • アフリカツメガエルゲノムの遺伝子進化: 脊椎動物の誕生に挑む Invited

    中央大学附属高校・ステップ講座  2017.10 

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  • X.laevisのゲノム解析:バイオインフォマティクスを始めるために

    福井彰雅

    第11回日本ツメガエル研究集会  2017.9 

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  • ツメガエルケモカイン関連遺伝子の網羅的解析

    福井彰雅, 松波雅俊

    日本動物学会第88回大会  2017.9 

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  • アフリカツメガエルゲノムプロジェクト: 異質四倍体ゲノムの解析 Invited

    福井彰雅

    琉球大学公開セミナー  2017.9 

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  • ツメガエルケモカイン遺伝子の網羅的解析:異質四倍体化での遺伝子進化

    福井彰雅, 松波雅俊

    第3回次世代両生類研究会  2017.8 

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  • ツメガエル中内胚葉細胞は胞胚腔蓋をハプトタキシスにより移動する

    福井彰雅

    7th Tokyo Vertebrate Morphology Meeting  2017.7 

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  • ゲノムが倍加する意味 Invited

    福井彰雅

    慶応大学SFC・概念構築  2017.5 

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  • ツメガエル中内胚葉細胞は胞胚腔蓋をハプトタキシスにより移動する

    遠藤綾乃

    第38回日本分子生物学会年会  2016.11 

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  • 細胞間接着に関与するCXCR7-C末端領域の解析

    越場貴能, 佐々木直樹, 古澤和也, 福井彰雅

    第38回日本分子生物学会年会  2016.11 

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  • ツメガエルケモカイン遺伝子の網羅的解析:異質四倍体化による遺伝子進化

    福井彰雅, 松波雅俊

    第38回日本分子生物学会年会  2016.11 

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  • ツメガエルケモカイン及びその受容体の遺伝子構造解析

    福井彰雅, 松波雅俊

    第10回日本ツメガエル研究集会  2016.11 

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  • ケモカイン濃度勾配の定量的解析

    福井彰雅

    日本動物学会第85回大会  2015.9 

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  • CXCR7情報伝達経路による細胞間接着機構の研究

    小山晶平

    第37回日本分子生物学会年会  2014.11 

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  • ケモカイン勾配形成とその定量的研究

    泉野奈都子

    第37回日本分子生物学会年会  2014.11 

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  • 組織常在型M2マクロファージ細胞の培養系の確立と間葉系細胞への影響について

    藤岡明博

    第37回日本分子生物学会年会  2014.11 

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  • アフリカツメガエル初期胚におけるnodal関連遺伝子Xnr5の3D-FISH

    山廣 傑, 古澤和也, 佐々木直樹, 福井彰雅

    第37回日本分子生物学会年会  2014.11 

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  • RNF43遺伝子のミスセンス変異によるWnt/βcatenin経路のポジティブフィードバックループ形成

    築山忠維

    第37回日本分子生物学会年会  2014.11 

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  • SDF-1情報伝達経路による細胞の接着制御

    第7回日本ツメガエル研究集会  2013.9 

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  • CXCR7による原腸胚期中内胚葉細胞の細胞間接着制御機構

    福井彰雅, 古澤和也, 佐々木直樹

    日本動物学会第84回大会  2013.9 

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  • RNF43はCanonical/noncanonical Wntシグナルの両方を抑制する

    築山忠維

    第35回日本分子生物学会  2012.12 

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  • xSDF-1情報伝達関連因子の発現パターンについて

    第5回日本ツメガエル研究集会  2011.10 

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  • アフリカツメガエル原腸胚期でのxCXCR7の機能について

    桝田啓太

    第5回日本ツメガエル研究集会  2011.10 

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  • ツメガエルの原腸形成に見られる細胞集団運動 ―SDF-1情報伝達経路による細胞運動制御機構―

    日本動物学会第82回大会  2011.9 

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  • ラット組織におけるマクロファージ多型の分布と局在

    佐藤真希

    日本動物学会第82回大会  2011.9 

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  • アフリカツメガエル原腸胚期でのxCXCR7の機能について

    桝田啓太

    日本動物学会第82回大会  2011.9 

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  • アクチビン初期応答遺伝子の染色体マッピング

    森晴香 ほ

    日本動物学会第82回大会  2011.9 

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  • アフリカツメガエルPAFRの強制発現及び翻訳阻害実験による原腸陥入運動阻害の観察

    井上直樹, 古澤和也, 佐々木直樹, 福井彰雅

    日本動物学会第82回大会  2011.9 

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  • 細胞運動制御におけるBrachyury遺伝子の進化的に保存された役割の研究

    山田温子

    日本動物学会第82回大会  2011.9 

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  • ヒストンH1の過剰発現により誘導される遺伝子のツメガエル染色体座位の同定

    山廣傑 ほ

    日本動物学会第81回大会  2010.9 

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  • ツメガエル中内胚葉性細胞の移動に伴うCa++の変化の観察

    佐藤巧翔

    日本動物学会第81回大会  2010.9 

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  • アフリカツメガエルCXCR7による原腸陥入制御について

    桝田啓太

    日本動物学会第81回大会  2010.9 

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  • 原腸陥入期におけるSDF-1関連遺伝子の発現領域の観察

    長田朋子

    日本動物学会第81回大会  2010.9 

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  • アフリカツメガエル原腸胚期でのPAFR様遺伝子の発現と機能について

    井上直樹, 古澤和也, 佐々木直樹, 福井彰雅

    日本動物学会第81回大会  2010.9 

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  • ラット脂肪由来間質細胞の骨分化における力学刺激の影響

    川村亜沙美, 古澤和也, 佐々木直樹, 福井彰雅

    第32回日本分子生物学会年会  2009.12 

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  • 原腸陥入期におけるツメガエルCXCR7の局在

    第3回日本ツメガエル研究集会  2009.10 

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  • ツメガエル原腸陥入期におけるケモカインSDF-1およびその受容体の分布と機能について

    福井彰雅, 桝田啓太, 長田朋子, 佐々木直樹

    日本動物学会第80回大会  2009.9 

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  • ツメガエルケモカイン受容体の原腸陥入における役割について

    田中麻里子, 佐々木直樹, 福井彰雅

    第31回日本分子生物学会年会第81回日本生化学会 合同大会  2008.12 

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  • ラット脂肪由来間質細胞(ASC)の骨分化能の研究

    川村亜沙美, 佐々木直樹, 福井彰雅

    日本分子生物学会第8回春期シンポジウム  2008.5 

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  • アフリカツメガエル初期胚におけるNucelosome assembly protein-1 (NAP-1)の分布

    山廣 傑, 浦 聖恵, 佐々木 直樹, 福井彰雅

    日本分子生物学会第8回春期シンポジウム  2008.5 

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  • ΦC31インテグラーゼを用いたトランスジェニックイモリ作製技術の検討

    河合 淳一郎, 佐々木 直樹, 福井彰雅

    日本分子生物学会第8回春期シンポジウム  2008.5 

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  • 未分化細胞から誘導した血管内皮細胞を用いた新規血管関連遺伝子の単離

    長嶺憲太郎

    第30回日本分子生物学会年会第80回日本生化学会 合同大会  2007.12 

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  • Leading Edge Mesoderm(先行中胚葉)の運動におけるCXCR4/SDF-1系の役割について

    第1回日本ツメガエル研究集会  2007.8 

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  • 骨の力学特性に及ぼすカルシウムイオン溶出の影響

    野添 勉, 福井彰雅, 佐々木直樹

    日本バイオレオロジー学会  2007.6 

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  • 培養骨芽細胞による石灰化に及ぼす力学刺激の影響

    伊東大輔, 福井彰雅, 佐々木直樹

    日本バイオレオロジー学会  2007.6 

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  • Ia型BMP受容体欠損マウス骨の構造と性質

    斉藤 徹 ほ

    日本バイオレオロジー学会誌  2007.6 

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  • 東京大学における基礎生命科学実験:新カリキュラムに向けた取組み

    箕浦高子

    日本動物学会第77回大会  2006.9 

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  • マイクロアレイを用いたクリノスタットローテーション下のA6細胞における遺伝子発現変化の解析

    生澤昌之, 福井彰雅, 浅島 誠

    日本宇宙生物学会第19回大会  2006.9 

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  • アクチビンAは海馬で樹状突起スパインを調節する

    三橋賢司

    第43回日本生物物理学会年会  2005.11 

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  • CXCR4/SDF-1系のツメガエル原腸陥入時の役割について

    福井彰雅

    日本動物学会第76回大会  2005.9 

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  • ツメガエル原腸陥入時のSDF-1/CXCR4の作用について

    福井彰雅, 本間幹啓, 浅島 誠

    日本発生生物学会第38回大会  2005.6 

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  • ツメガエル初期胚におけるWntシグナル伝達経路関連遺伝子Daple-likeの機能解析

    道上達男

    日本発生生物学会第38回大会  2005.6 

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  • 宇宙実験を想定した試薬保存確認試験

    矢野幸子

    宇宙利用シンポジウム 第21回  2005.3 

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  • 宇宙ステーション冷凍庫を模擬した試料冷却試験

    矢野幸子

    宇宙利用シンポジウム 第21回  2005.3 

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  • アクチビン処理により変動する遺伝子の解析

    福井彰雅, 本間幹啓, 浅島 誠

    日本動物学会第75回大会  2004.9 

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  • アフリカツメガエルにおけるDuplinの機能解析

    福井彰雅, 浅島 誠

    日本動物学会第73回大会  2002.9 

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  • Axin変異体のツメガエル軸形成における影響について

    福井彰雅, 岸田昭世, 菊池 章, 浅島 誠

    日本動物学会第71回大会  2000.9 

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  • XenopusのaPKCとASIP/PAR‐3は卵成熟に伴って動物半球への非対称分布を示す

    中谷雅明, ほか

    第22回日本分子生物学会年会  1999.12 

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  • β-カテニンと結合する新規蛋白質 Duplinの同定および機能解析

    坂本郁夫, ほか

    第22回日本分子生物学会年会  1999.12 

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  • XenopusのaPKCとASIP/PAR‐3は卵成熟に伴って動物半球への非対称分布を示す

    中谷雅明, ほか

    第21回日本分子生物学会年会  1998.12 

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  • アクチビンおよびフォリスタチンとビテロゲニンの分子間相互作用の解析

    福井彰雅, 浅島 誠

    日本発生生物学会第31回大会  1998.5 

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  • アクチビン及びフォリスタチンのツメガエル卵母細胞への取り込みについて

    福井彰雅, 駒崎伸二, 浅島 誠

    日本動物学会第68回大会  1997.10 

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  • 中胚葉誘導因子であるアクチビンの卵母細胞への取り込み機構について

    福井彰雅, 岡林浩嗣, 杉野 弘, 浅島 誠

    第19回日本分子生物学会年会・第69回日本生化学会大会 合同年会  1996.8 

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  • 両生類の初期胚での形づくりと細胞分化の分子的制御

    浅島 誠 ほ

    第18回日本分子生物学会年会  1995.12 

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  • ツメガエル初期胚でのアクチビン・フォリスタチンの役割

    福井彰雅

    日本動物学会第64回大会  1993.11 

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  • ツメガエル初期胚からのアクチビン・フォリスタチンの精製

    福井彰雅

    日本発生生物学会第26回大会  1993.5 

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  • ツメガエル由来の培養細胞からのフォリスタチンの精製

    福井彰雅

    日本発生生物学会第25回大会  1992.5 

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  • ツメガエル卵へのフォリスタチン(アクチビン結合タンパク質)の注入

    岩男めぐみ, 福井彰雅, 内山英穂, 浅島 誠

    日本動物学会第62回大会  1991.10 

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  • ツメガエル卵巣のフォリスタチンの精製とその性質

    福井彰雅

    日本動物学会第62回大会  1991.10 

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  • ツメガエル細胞でのアクチビンA(=EDF)および関連タンパク質の研究

    福井彰雅, 浅島 誠, 西松伸一郎, 上野直人

    日本動物学会第61回大会  1990.10 

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  • アフリカツメガエル腎臓由来のA6細胞の形態的特徴について

    福井彰雅, 浅島 誠

    日本動物学会第60回大会  1989.10 

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Research Projects

  • Construction of morphogenetic mechanism analysis system for shape change and mechanical stimulation by cell origami folding technique

    Grant number:17H03194  2017.4 - 2021.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)  Hokkaido University

    Shigetomi Kaori

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    Grant amount: \18070000 ( Direct Cost: \13900000 、 Indirect Cost: \4170000 )

    In this study, we use microfabrication technology and folding technology of origami engineering to create microplates that serve as scaffolds for cells, and by folding the cells after culturing them, the shape changes due to the folding of the cells during the process of morphogenesis. We aimed to elucidate the relationship between mechanical stimulation and the mechanism of morphogenesis in vivo. During the research period, we produced a magnetic microplate, cells were cultured on the plate, and then the microplate was folded and unfolded using a magnetic field under a microscope, and the shape of the cells was successfully controlled three-dimensionally. By using a magnetic material, it can be driven in a non-contact manner, so it is possible to change the shape of cells without the need for wiring.

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  • Regulation of collective cell movement by chemokine receptor CXCR7

    Grant number:23590215  2011 - 2013

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)  Hokkaido University

    FUKUI AKIMASA

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    Grant amount: \5200000 ( Direct Cost: \4000000 、 Indirect Cost: \1200000 )

    Collective cell movement is a widely observed biological event, and not a few parts of that are controlled by chemoattractants. A chemokine stromal cell-derived factor-1 (SDF-1), also known as CXCL12, functions as chemoattractant through its G-protein coupled receptor CXCR4. CXCR7, which was identified as a second receptor for SDF-1, has been focused since its novel roles have been reported as controlling cell migration to inhibit the signaling by sequestrating the SDF-1 and to signal via non-G-protein signaling pathway. In the present study, several lines of experiment suggest xCXCR7/SDF-1-dependent novel function that CXCR7 signaling modulates intercellular adhesion.

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  • ツメガエルデュプリンの機能解析

    Grant number:14780556  2002 - 2003

    日本学術振興会  科学研究費助成事業  若手研究(B)  東京大学

    福井 彰雅

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    Grant amount: \2400000 ( Direct Cost: \2400000 )

    duplin-short form(Xduls)の単離のため、3-prime RACEにより得られた情報をもとにspecific probeを設計し、これを用いて逆転写したcDNAをライブラリー化してクローニングを行った。約130000クローンの内一つがpositiveであり、これがXdulsの全長配列を含んでいた。新たに分かったXdulsのN末端側はすでに得られたツメガエルduplinと非常に高い相同性を示した。このXdulsとGFPのfusion constructを作製し、ツメガエル卵へ強制発現させて細胞内の局在を調べた結果、核にシグナルが見られた。これよりXdulsは核内移行することが分かった。XdulsのmRNAを合成し、4細胞期胚の異なる領域に微量注入してその結果を観察した。背側植物極へ注入した際は、頭部構造の欠損が見られた。背側動物極側へ注入した際、原腸陥入異常が見られ、背側に屈曲した幼生がみられた。腹側へXwnt-8と同時に注入することにより、Xwnt-8による2次軸形成を抑制した。この遺伝子の翻訳を阻害すると考えられるmorpholino antisense oligomerを設計し、背側動物極側へ注入した胚では、頭部構造の異常が見られた。また、Xdulsの作用部位を調べるため、この遺伝子のさまざまな欠失変異体を作製し、その活性を調べた。その結果、C末端側の塩基性アミノ酸クラスターがこの因子の活性に重要であることがわかった。以前の結果と共に、Xdulsは原腸胚期から発現が始まり、初期発生中には頭部形成と細胞の運動に関与すること、その活性には塩基性アミノ酸クラスターが重要であることが示された。

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  • 新規のWntシグナル伝達分子による形態形成の制御機構の解析

    Grant number:14034239  2002 - 2003

    日本学術振興会  科学研究費助成事業  特定領域研究  広島大学

    岸田 昭世, 菊池 章, 福井 彰雅

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    Grant amount: \6600000 ( Direct Cost: \6600000 )

    Wntシグナルは初期発生時の体軸形成や細胞の増殖や分化などを制御する。Wntシグナルは細胞内でβ-カテニン経路とPCP(平面内細胞極性)経路、Ca^<2+>経路の3つに分岐する。DvlはWntシグナルをβ-カテニン経路とPCP経路に分岐点に当たる分子で、GSK-3βによるβ-カテニンのリン酸化と分解を抑制する一方、ショウジョウバエでの解析から、PCP経路によってJunキナーゼやRhoキナーゼを介して形態形成を制御することが見出されつつあるが、神経系細胞におけるPCP経路の作用は明らかでなかった。
    そこで今年度は、哺乳動物細胞におけるWntやDvl、Rhoキナーゼの作用を検討した。COS細胞において、Junキナーゼ活性化に必須のDEP領域を含まないDvl変異体がRhoキナーゼを活性化する事から、DvlはJunキナーゼとRhoキナーゼを異なった領域を介して活性化する可能性が示された.Wnt-1やWnt-3a、DvlによるRhoキナーゼの活性化はRhoキナーゼのRho結合ドメインを共発現することにより抑制されたことから、この活性化にはRhoが関与することが明らかとなった。Wnt-1やDvlを発現すると褐色細胞腫由来のPC12細胞でのNGF依存性の神経突起伸張や神経芽細胞腫由来N1E-115細胞での血清除去刺激による神経突起伸張が抑制されだが、この現象はRhoキナーゼ阻害剤により解除された。さらに、RhotekinのRho結合領域を用いたpull-down assayにより、Dvlを発現させたPC12細胞ではRhoが活性化することを見出した。高度に精製したWnt-3aでPC12細胞を処理することによりRhoキナーゼの活性化が認められた。したがって、Wnt-3aやDvlのシグナルは哺乳動物の神経系細胞において、Rhoキナーゼを介して神経突起の伸張や退縮に関わる可能性が示唆された。

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  • ツメガエル卵母細胞におけるアクチビン情報伝達の解析

    Grant number:12780551  2000 - 2001

    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

    福井 彰雅

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    Grant amount: \2100000 ( Direct Cost: \2100000 )

    これまでの研究により、中胚葉誘導因子として働くアクチビンが受精前後の卵中にすでにタンパク質として存在していること、さらにアクチビンが卵母細胞へ取り込まれることを明らかにしてきた。アクチビンは発生過程でも働くが、細胞の生存や維持に効果があることも知られている。この研究では、取り込まれたアクチビンがどのように卵母細胞に作用しているかを調べるため、まずその受容体の局在を調べた。免疫抗体染色法による観察では、IB型アクチビン受容体はツメガエル卵母細胞内のミトコンドリア集合体(バルビニア小体)と呼ばれる構造に局在がみられた。凍結切片を用いた免疫電子顕微鏡法による観察では、アクチビン受容体IB抗体によるシグナルはミトコンドリア上では見られず、滑面小胞体で見られた。ミトコンドリア集合体には、ミトコンドリア、小胞体、核由来物質などが存在することが知られているが、実験結果より、アクチビン受容体はミトコンドリアではなく小胞体に存在することがわかった。抗体染色により得られた像がアクチビン受容体であることを確認するため、卵母細胞を試料とし、さまざまな調製をおこなった後にウェスタンブロット法により分子量の同定を試みたが、確認できなかった。これは卵母細胞中のアクチビン受容体分子数が少ないことが原因と考え、IB型アクチビン受容体のmRNAを胚に注入し、強制発現させたところ、60kDa付近にバンドが見られ、用いた抗体がIB型アクチビン受容体を認識していることが確認された。また、アクチビン情報伝達系のメディエーター分子として知られるSmadに対する抗体を作成し、その局在を調べたところ核小体にそのシグナルが見られた。これらのことより、アクチビン情報伝達系が卵母細胞内で機能している可能性が示唆される。

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  • Moleculer mechanisms of cell differentiation and morphogenesis during early animal development

    Grant number:07408023  1995 - 1997

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (A)  The University of Tokyo

    ASASHIMA Makoto, FUKUI Akimasa

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    Grant amount: \35600000 ( Direct Cost: \35600000 )

    We got many important results about the morphogenesis and cell differentiation using the amphibian embryos. (1) After the treatment of activin A on animal caps, we made subtraction methods to clone the neurogenesis-related genes. And we cloned and analyzed the new genes such as Xran, XHMG-2, XIRF-6, XFRP,XGS,XNLRR-1. (2) In vitro system, we succeeded to make the head structure and trunk-tail structures. These structures were made with sandwith methods, in which the activin-treated ectoderm was combined. Depend on the pre-culture time, the induced structures were different. (3) Some kinds of genes which were related with pronephros (kidney) formation were cloned and analyzed. These genes were Na-K-ATPase alpha-subunit, CIRP,XFKBP,and the expression patterns of these genes are examined. (4) Incorporations of activin, follistatin and vitellogenin were examined using the labeling of these substance with ^<125>I and gold colloid particles. (5) Treatment with high concentration of activin to animal cap induced the beating heart, and expressed many kinds of endodermal marker genes. (6) Vitellogenin receptor during oogenesis was also cloned and analyzed. Above these data were very important to understand the cell differentiation and morphogenesis at the molecular level during animal development.

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  • 両生類初期発生におけるTGF/s関連タンパク質の作用機序

    Grant number:08780700  1996    

    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

    福井 彰雅

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    Grant amount: \1000000 ( Direct Cost: \1000000 )

    両生類初期胚では中胚葉誘導、神経誘導といった一連の誘導現象にアクチビンなどのTGFβスーパーファミリーに属するタンパク質因子が強く関与していることが報告されている。すでに初期胚中にアクチビンおよびフォリスタチンがタンパク質の形で存在していることは証明したが、これらの分子の蓄積機構については分かっていない。この研究ではアクチビンの活性化機構に先立ち、まずアクチビンおよびフォリスタチンの卵母細胞への蓄積機構について卵母細胞のin vitro培養系を用いて調べた。
    アクチビンA、Bおよびフォリスタチンを^<125>lでラベルし、これらの卵母細胞への取り込みを観察した。アルビノのツメガエルを用い、^<125>lラベルしたアクチビンおよびフォリスタチンを取り込ませた卵母細胞の切片を作製し、このオートラジオグラフィをとった。この結果、アクチビンやフォリスタチンは確実に卵母細胞の細胞質中に移行することがわかった。この時、コントロールのBSAの取り込みは全く観察されなかった。この結果より、アクチビンおよびフォリスタチンが卵母細胞内に取り込まれることが判明したが、これらの細胞質中での局在に関しては不明である。そこで、アクチビンおよびフォリスタチンを金コロイドでラベルし、この取り込みを調べ、卵母細胞内への蓄積を電子顕微鏡を用いて観察した。この結果、金コロイドは卵母細胞中へ取り込まれ、最終的に卵黄顆粒に移行することが示された。これらの結果はアクチビンおよびフォリスタチンが卵母細胞外から取り込まれ、卵母細胞中の卵黄顆粒に蓄積されることを示唆する。今後はこれら卵中に蓄積されたアクチビンの活性化機構について研究を進める予定である。

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  • 2018.5 -  

    ナショナルバイオリソースプロジェクト(NBRP)   AMED ナショナルバイオリソースプロジェクト(NBRP) ネッタイツメガエル運営委員会委員