2024/04/05 更新

写真a

シバタ ユウキ
柴田 侑毅
Shibata Yuki
所属
医学部 生物学 講師
職名
講師

研究キーワード

  • ゲノム編集

  • 変態

  • 再生

  • トランスジェネシス

  • 両生類

  • 甲状腺ホルモン(TH)

  • 甲状腺ホルモン受容体

研究分野

  • ライフサイエンス / 発生生物学  / 両生類 変態 再生 ゲノム編集 甲状腺ホルモン受容体

学歴

  • 静岡大学創造科学技術大学院   自然科学系教育部   バイオサイエンス

    2012年4月 - 2015年3月

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    国名: 日本国

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  • 静岡大学   大学院理学研究科   生物科学専攻

    2010年4月 - 2012年3月

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  • 静岡大学   理学部   生物科学科

    2006年4月 - 2010年3月

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経歴

  • 日本医科大学   生物学教室   講師

    2022年8月 - 現在

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  • 基礎生物学研究所   新規モデル生物開発センター   特任研究員

    2021年4月 - 2022年8月

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    国名:日本国

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  • National Institute of Health (NIH)   NICHD   JSPS 海外特別研究員NIH (海特NIH)

    2017年3月 - 2019年2月

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    国名:アメリカ合衆国

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  • National Institutes of Health (NIH)   Eunice Kennedy Shriver National Institute Child Health and Human Development (NICHD)   Visiting fellow

    2015年7月 - 2021年3月

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    国名:アメリカ合衆国

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  • 静岡大学   理学部 生物科学科   博士研究員

    2015年4月 - 2015年6月

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    国名:日本国

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  • 静岡大学   創造科学技術大学院   特別研究員 (DC2)

    2014年4月 - 2015年3月

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    国名:日本国

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▼全件表示

所属学協会

  • アメリカ甲状腺学会

    2017年 - 現在

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  • 日本比較内分泌学会

    2010年 - 現在

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  • 日本動物学会

    2010年 - 現在

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委員歴

  • 日本比較内分泌学会   若手交流企画委員  

    2021年 - 現在   

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    団体区分:学協会

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論文

  • Protocols for transgenesis at a safe harbor site in the Xenopus laevis genome using CRISPR-Cas9 査読

    Yuki Shibata, Akinori Okumura, Makoto Mochii, Ken-ichi T. Suzuki

    STAR Protocols   4 ( 3 )   102382 - 102382   2023年9月

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    担当区分:筆頭著者   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    添付ファイル: 2782.pdf

    DOI: 10.1016/j.xpro.2023.102382

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  • CRISPR/Cas9-based simple transgenesis in Xenopus laevis 査読 国際誌

    Yuki Shibata, Miyuki Suzuki, Nao Hirose, Ayuko Takayama, Chiaki Sanbo, Takeshi Inoue, Yoshihiko Umesono, Kiyokazu Agata, Naoto Ueno, Ken-ichi T. Suzuki, Makoto Mochii

    Developmental Biology   489   76 - 83   2022年6月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    Transgenic techniques have greatly increased our understanding of the transcriptional regulation of target genes through live reporter imaging, as well as the spatiotemporal function of a gene using loss- and gain-of-function constructs. In Xenopus species, two well-established transgenic methods, restriction enzyme-mediated integration and I-SceI meganuclease-mediated transgenesis, have been used to generate transgenic animals. However, donor plasmids are randomly integrated into the Xenopus genome in both methods. Here, we established a new and simple targeted transgenesis technique based on CRISPR/Cas9 in Xenopus laevis. In this method, Cas9 ribonucleoprotein (RNP) targeting a putative harbor site (the transforming growth factor beta receptor 2-like (tgfbr2l) locus) and a preset donor plasmid DNA were co-injected into the one-cell stage embryos of X. laevis. Approximately 10% of faithful reporter expression was detected in F0 crispants in a promoter/enhancer-specific manner. Importantly, efficient germline transmission and stable transgene expression were observed in the F1 offspring. The simplicity of this method only required preparation of a donor vector containing the tgfbr2l genome fragment and Cas9 RNP targeting this site, which are common experimental procedures used in Xenopus laboratories. Our improved technique allows the simple generation of transgenic X. laevis, so is expected to become a powerful tool for reporter assay and gene function analysis.

    DOI: 10.1016/j.ydbio.2022.06.001

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  • Thyroid Hormone Receptor Is Essential for Larval Epithelial Apoptosis and Adult Epithelial Stem Cell Development but Not Adult Intestinal Morphogenesis during Xenopus tropicalis Metamorphosis. 査読 国際誌

    Yuki Shibata, Yuta Tanizaki, Hongen Zhang, Hangnoh Lee, Mary Dasso, Yun-Bo Shi

    Cells   10 ( 3 )   2021年3月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Vertebrate postembryonic development is regulated by thyroid hormone (T3). Of particular interest is anuran metamorphosis, which offers several unique advantages for studying the role of T3 and its two nuclear receptor genes, TRα and TRβ, during postembryonic development. We have recently generated TR double knockout (TRDKO) Xenopus tropicalis animals and reported that TR is essential for the completion of metamorphosis. Furthermore, TRDKO tadpoles are stalled at the climax of metamorphosis before eventual death. Here we show that TRDKO intestine lacked larval epithelial cell death and adult stem cell formation/proliferation during natural metamorphosis. Interestingly, TRDKO tadpole intestine had premature formation of adult-like epithelial folds and muscle development. In addition, T3 treatment of premetamorphic TRDKO tadpoles failed to induce any metamorphic changes in the intestine. Furthermore, RNA-seq analysis revealed that TRDKO altered the expression of many genes in biological pathways such as Wnt signaling and the cell cycle that likely underlay the inhibition of larval epithelial cell death and adult stem cell development caused by removing both TR genes. Our data suggest that liganded TR is required for larval epithelial cell degeneration and adult stem cell formation, whereas unliganded TR prevents precocious adult tissue morphogenesis such as smooth-muscle development and epithelial folding.

    DOI: 10.3390/cells10030536

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  • Organ-Specific Requirements for Thyroid Hormone Receptor Ensure Temporal Coordination of Tissue-Specific Transformations and Completion of Xenopus Metamorphosis. 査読 国際誌

    Yuki Shibata, Luan Wen, Morihiro Okada, Yun-Bo Shi

    Thyroid : official journal of the American Thyroid Association   30 ( 2 )   300 - 313   2020年2月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Background: Thyroid hormone (triiodothyronine [T3]) is essential for the development throughout vertebrates. Anuran metamorphosis mimics mammalian postembryonic development, a period around birth when plasma T3 level peaks and many organs/tissues mature into their adult forms. Compared with the uterus-enclosed mammalian embryos, tadpoles can be easily manipulated to study the roles of T3 and T3 receptors (TRs) in tissue remodeling and adult organ development. We and others have previously knocked out TRα or TRβ in the diploid anuran Xenopus tropicalis and reported distinct effects of the two receptor knockouts on metamorphosis. However, animals lacking either TRα or TRβ can complete metamorphosis and develop into reproductive adults. Methods: We have generated TRα and TRβ double knockout animals and carried out molecular and morphological analyses to determine if TR is required for Xenopus development. Results: We found that the TR double knockout tadpoles do not respond to T3, supporting the view that there are no other TR genes in X. tropicalis and that TR is essential for mediating the effects of T3 in vivo. Surprisingly, the double knockout tadpoles are able to initiate metamorphosis and accomplish many metamorphic changes, such as limb development. However, all double knockout tadpoles stall and eventually die at stage 61, the climax of metamorphosis, before tail resorption takes place. Analyses of the knockout tadpoles at stage 61 revealed various developmental abnormalities, including precocious ossification and extra vertebrae. Conclusions: Our data indicate that TRs are not required for the initiation of metamorphosis but is essential for the completion of metamorphosis. Furthermore, the differential effects of TR knockout on different organs/tissues suggest tissue-specific roles for TR to control temporal coordination and progression of metamorphosis in various organs.

    DOI: 10.1089/thy.2019.0366

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  • Fgf10 mutant newts regenerate normal hindlimbs despite severe developmental defects 査読

    Miyuki Suzuki, Akinori Okumura, Akane Chihara, Yuki Shibata, Tetsuya Endo, Machiko Teramoto, Kiyokazu Agata, Marianne E. Bronner, Ken-ichi T. Suzuki

    Proceedings of the National Academy of Sciences   121 ( 11 )   2024年3月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Proceedings of the National Academy of Sciences  

    In amniote limbs, Fibroblast Growth Factor 10 (FGF10) is essential for limb development, but whether this function is broadly conserved in tetrapods and/or involved in adult limb regeneration remains unknown. To tackle this question, we established Fgf10 mutant lines in the newt Pleurodeles waltl which has amazing regenerative ability. While Fgf10 mutant forelimbs develop normally, the hindlimbs fail to develop and downregulate FGF target genes. Despite these developmental defects, Fgf10 mutants were able to regenerate normal hindlimbs rather than recapitulating the embryonic phenotype. Together, our results demonstrate an important role for FGF10 in hindlimb formation, but little or no function in regeneration, suggesting that different mechanisms operate during limb regeneration versus development.

    DOI: 10.1073/pnas.2314911121

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  • Ontogenetic Expression of Aquaporins in the Kidney and Urinary Bladder of the Japanese Tree Frog, Dryophytes japonicus 招待 査読

    Masatoshi Hibino, Ryota Aoki, Duy Anh Ha, Haruna Sano, Shiori Yamashita, Haruto Ogasawara, Kazuma Nishio, Kohei Kotake, Md. Main Uddin Mamun, Reiko Okada, Yuki Shibata, Masakazu Suzuki

    Zoological Science   41 ( 1 )   2024年2月

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    担当区分:責任著者   掲載種別:研究論文(学術雑誌)   出版者・発行元:Zoological Society of Japan  

    DOI: 10.2108/zs230069

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  • Simplifying Genotyping of Mutants from Genome Editing with a Parallel qPCR-Based iGenotype Index

    Liezhen Fu, Shouhong Wang, Lusha Liu, Yuki Shibata, Morihiro Okada, Nga Luu, Yun-Bo Shi

    Cells   2024年1月

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    掲載種別:研究論文(学術雑誌)  

    DOI: 10.3390/cells13030247

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  • Thyroid Hormone-Activated Signaling Pathways are Essential for Development of Intestinal Stem Cells 査読

    Kenta Fujimoto, Yuki Shibata, Takashi Hasebe

    Journal of Nippon Medical School   90 ( 3 )   246 - 252   2023年6月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Medical Association of Nippon Medical School  

    DOI: 10.1272/jnms.jnms.2023_90-308

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  • Cell cycle activation in thyroid hormone-induced apoptosis and stem cell development during Xenopus intestinal metamorphosis

    Yuta Tanizaki, Yuki Shibata, Wonho Na, Yun-Bo Shi

    Frontiers in Endocrinology   14   2023年5月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Frontiers Media SA  

    Amphibian metamorphosis resembles mammalian postembryonic development, a period around birth when many organs mature into their adult forms and when plasma thyroid hormone (T3) concentration peaks. T3 plays a causative role for amphibian metamorphosis. This and its independence from maternal influence make metamorphosis of amphibians, particularly anurans such as pseudo-tetraploid Xenopus laevis and its highly related diploid species Xenopus tropicalis, an excellent model to investigate how T3 regulates adult organ development. Studies on intestinal remodeling, a process that involves degeneration of larval epithelium via apoptosis and de novo formation of adult stem cells followed by their proliferation and differentiation to form the adult epithelium, have revealed important molecular insights on T3 regulation of cell fate during development. Here, we review some evidence suggesting that T3-induced activation of cell cycle program is important for T3-induced larval epithelial cell death and de novo formation of adult intestinal stem cells.

    DOI: 10.3389/fendo.2023.1184013

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  • Competitive PCR with dual fluorescent primers enhances the specificity and reproducibility of genotyping animals generated from genome editing 査読

    Liezhen Fu, Emily Ma, Morihiro Okada, Yuki Shibata, Yun-Bo Shi

    Cell & Bioscience   13 ( 1 )   2023年5月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer Science and Business Media LLC  

    Abstract

    Targeted genome editing is a powerful tool for studying gene function in almost every aspect of biological and pathological processes. The most widely used genome editing approach is to introduce engineered endonucleases or CRISPR/Cas system into cells or fertilized eggs to generate double-strand DNA breaks within the targeted region, leading to DNA repair through homologous recombination or non-homologous end joining (NHEJ). DNA repair through NHEJ mechanism is an error-prone process that often results in point mutations or stretches of indels (insertions and deletions) within the targeted region. Such mutations in embryos are germline transmissible, thus providing an easy means to generate organisms with gene mutations. However, point mutations and short indels present difficulty for genotyping, often requiring labor intensive sequencing to obtain reliable results. Here, we developed a single-tube competitive PCR assay with dual fluorescent primers that allowed simple and reliable genotyping. While we used Xenopus tropicalis as a model organism, the approach should be applicable to genotyping of any organisms.

    DOI: 10.1186/s13578-023-01042-2

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    その他リンク: https://link.springer.com/article/10.1186/s13578-023-01042-2/fulltext.html

  • Comparative Analysis of Transcriptome Profiles Reveals Distinct and Organ-Dependent Genomic and Nongenomic Actions of Thyroid Hormone in Xenopus tropicalis Tadpoles 査読

    Shouhong Wang, Yuki Shibata, Yuta Tanizaki, Hongen Zhang, Wei Yan, Liezhen Fu, Yun-Bo Shi

    Thyroid   2023年4月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Mary Ann Liebert Inc  

    DOI: 10.1089/thy.2022.0469

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  • Thyroid hormone receptor knockout prevents the loss of Xenopus tail regeneration capacity at metamorphic climax 査読

    Shouhong Wang, Yuki Shibata, Liezhen Fu, Yuta Tanizaki, Nga Luu, Lingyu Bao, Zhaoyi Peng, Yun-Bo Shi

    Cell & Bioscience   13 ( 1 )   2023年2月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer Science and Business Media LLC  

    Abstract

    Background

    Animal regeneration is the natural process of replacing or restoring damaged or missing cells, tissues, organs, and even entire body to full function. Studies in mammals have revealed that many organs lose regenerative ability soon after birth when thyroid hormone (T3) level is high. This suggests that T3 play an important role in organ regeneration. Intriguingly, plasma T3 level peaks during amphibian metamorphosis, which is very similar to postembryonic development in humans. In addition, many organs, such as heart and tail, also lose their regenerative ability during metamorphosis. These make frogs as a good model to address how the organs gradually lose their regenerative ability during development and what roles T3 may play in this. Early tail regeneration studies have been done mainly in the tetraploid Xenopus laevis (X. laevis), which is difficult for gene knockout studies. Here we use the highly related but diploid anuran X. tropicalis to investigate the role of T3 signaling in tail regeneration with gene knockout approaches.

    Results

    We discovered that X. tropicalis tadpoles could regenerate their tail from premetamorphic stages up to the climax stage 59 then lose regenerative capacity as tail resorption begins, just like what observed for X. laevis. To test the hypothesis that T3-induced metamorphic program inhibits tail regeneration, we used TR double knockout (TRDKO) tadpoles lacking both TRα and TRβ, the only two receptor genes in vertebrates, for tail regeneration studies. Our results showed that TRs were not necessary for tail regeneration at all stages. However, unlike wild type tadpoles, TRDKO tadpoles retained regenerative capacity at the climax stages 60/61, likely in part by increasing apoptosis at the early regenerative period and enhancing subsequent cell proliferation. In addition, TRDKO animals had higher levels of amputation-induced expression of many genes implicated to be important for tail regeneration, compared to the non-regenerative wild type tadpoles at stage 61. Finally, the high level of apoptosis in the remaining uncut portion of the tail as wild type tadpoles undergo tail resorption after stage 61 appeared to also contribute to the loss of regenerative ability.

    Conclusions

    Our findings for the first time revealed an evolutionary conservation in the loss of tail regeneration capacity at metamorphic climax between X. laevis and X. tropicalis. Our studies with molecular and genetic approaches demonstrated that TR-mediated, T3-induced gene regulation program is responsible not only for tail resorption but also for the loss of tail regeneration capacity. Further studies by using the model should uncover how T3 modulates the regenerative outcome and offer potential new avenues for regenerative medicines toward human patients.

    DOI: 10.1186/s13578-023-00989-6

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    その他リンク: https://link.springer.com/article/10.1186/s13578-023-00989-6/fulltext.html

  • Liver development during Xenopus tropicalis metamorphosis is controlled by T3-activation of WNT signaling. 査読

    Yuta Tanizaki, Shouhong Wang, Hongen Zhang, Yuki Shibata, Yun-Bo Shi

    iScience   106301 - 106301   2023年2月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    DOI: 10.1016/j.isci.2023.106301

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  • Organ‐specific effects on target binding due to knockout of thyroid hormone receptor α during Xenopus metamorphosis 査読

    Yuta Tanizaki, Hongen Zhang, Yuki Shibata, Yun‐Bo Shi

    Development, Growth & Differentiation   2023年1月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Wiley  

    DOI: 10.1111/dgd.12825

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  • Thyroid hormone receptor α controls larval intestinal epithelial cell death by regulating the CDK1 pathway 査読

    Yuta Tanizaki, Hongen Zhang, Yuki Shibata, Yun-Bo Shi

    Communications Biology   5 ( 1 )   2022年2月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer Science and Business Media LLC  

    <title>Abstract</title>Thyroid hormone (T3) regulates adult intestine development through T3 receptors (TRs). It is difficult to study TR function during postembryonic intestinal maturation in mammals due to maternal influence. We chose intestinal remodeling during <italic>Xenopus tropicalis</italic> metamorphosis as a model to study TR function in adult organ development. By using ChIP (chromatin immunoprecipitation)-Seq, we identified over 3000 TR-bound genes in the intestine of premetamorphic wild type or TRα (the major TR expressed during premetamorphosis)-knockout tadpoles. Surprisingly, cell cycle-related GO (gene ontology) terms and biological pathways were highly enriched among TR target genes even though the first major event during intestinal metamorphosis is larval epithelial cell death, and TRα knockout drastically reduced this enrichment. More importantly, treatment of tadpoles with cell cycle inhibitors blocked T3-induced intestinal remodeling, especially larval epithelial cell death, suggesting that TRα-dependent activation of cell cycle is important for T3-induced apoptosis during intestinal remodeling.

    DOI: 10.1038/s42003-022-03061-0

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    その他リンク: https://www.nature.com/articles/s42003-022-03061-0

  • Thyroid Hormone Receptor α Controls the Hind Limb Metamorphosis by Regulating Cell Proliferation and Wnt Signaling Pathways in Xenopus tropicalis 査読

    Yuta Tanizaki, Yuki Shibata, Hongen Zhang, Yun-Bo Shi

    International Journal of Molecular Sciences   23 ( 3 )   1223 - 1223   2022年1月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:MDPI AG  

    Thyroid hormone (T3) receptors (TRs) mediate T3 effects on vertebrate development. We have studied Xenopus tropicalis metamorphosis as a model for postembryonic human development and demonstrated that TRα knockout induces precocious hind limb development. To reveal the molecular pathways regulated by TRα during limb development, we performed chromatin immunoprecipitation- and RNA-sequencing on the hind limb of premetamorphic wild type and TRα knockout tadpoles, and identified over 700 TR-bound genes upregulated by T3 treatment in wild type but not TRα knockout tadpoles. Interestingly, most of these genes were expressed at higher levels in the hind limb of premetamorphic TRα knockout tadpoles than stage-matched wild-type tadpoles, suggesting their derepression upon TRα knockout. Bioinformatic analyses revealed that these genes were highly enriched with cell cycle and Wingless/Integrated (Wnt) signaling-related genes. Furthermore, cell cycle and Wnt signaling pathways were also highly enriched among genes bound by TR in wild type but not TRα knockout hind limb. These findings suggest that direct binding of TRα to target genes related to cell cycle and Wnt pathways is important for limb development: first preventing precocious hind limb formation by repressing these pathways as unliganded TR before metamorphosis and later promoting hind limb development during metamorphosis by mediating T3 activation of these pathways.

    DOI: 10.3390/ijms23031223

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  • 甲状腺ホルモン受容体αはCDK1経路の調節を介して幼生型小腸上皮細胞の細胞死を制御する

    祐太 谷崎, Hongen Zhang, 侑毅 柴田, Yun Bo Shi

    Comparative Endocrinology   48 ( 175 )   1 - 3   2022年

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Japan Society for Comparative Endocrinology  

    DOI: 10.5983/nl2008jsce.48.175_1_35

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  • Genetic basis for an evolutionary shift from ancestral preaxial to postaxial limb polarity in non-urodele vertebrates 査読

    Anna Trofka, Bau-Lin Huang, Jianjian Zhu, William F. Heinz, Valentin Magidson, Yuki Shibata, Yun-Bo Shi, Basile Tarchini, H. Scott Stadler, Mirindi Kabangu, Nour W. Al Haj Baddar, S. Randal Voss, Susan Mackem

    Current Biology   2021年11月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    DOI: 10.1016/j.cub.2021.09.010

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  • Analysis of Thyroid Hormone Receptor α-Knockout Tadpoles Reveals That the Activation of Cell Cycle Program Is Involved in Thyroid Hormone-Induced Larval Epithelial Cell Death and Adult Intestinal Stem Cell Development During Xenopus tropicalis Metamorphosis. 査読 国際誌

    Yuta Tanizaki, Yuki Shibata, Hongen Zhang, Yun-Bo Shi

    Thyroid : official journal of the American Thyroid Association   31 ( 1 )   128 - 142   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Background: There are two highly conserved thyroid hormone (triiodothyronine [T3]) receptor (TR) genes, TRα and TRβ, in all vertebrates, and the expression of TRα but not TRβ is activated earlier than T3 synthesis during development. In human, high levels of T3 are present during the several months around birth, and T3 deficiency during this period causes severe developmental abnormalities including skeletal and intestinal defects. It is, however, difficult to study this period in mammals as the embryos and neonates depend on maternal supply of nutrients for survival. However, Xenopus tropicalis undergoes a T3-dependent metamorphosis, which drastically changes essentially every organ in a tadpole. Of interest is intestinal remodeling, which involves near complete degeneration of the larval epithelium through apoptosis. Concurrently, adult intestinal stem cells are formed de novo and subsequently give rise to the self-renewing adult epithelial system, resembling intestinal maturation around birth in mammals. We have previously demonstrated that T3 signaling is essential for the formation of adult intestinal stem cells during metamorphosis. Methods: We studied the function of endogenous TRα in the tadpole intestine by using knockout animals and RNA-seq analysis. Results: We observed that removing endogenous TRα caused defects in intestinal remodeling, including drastically reduced larval epithelial cell death and adult intestinal stem cell proliferation. Using RNA-seq on intestinal RNA from premetamorphic wild-type and TRα-knockout tadpoles treated with or without T3 for one day, before any detectable T3-induced cell death and stem cell formation in the tadpole intestine, we identified more than 1500 genes, which were regulated by T3 treatment of the wild-type but not TRα-knockout tadpoles. Gene Ontology and biological pathway analyses revealed that surprisingly, these TRα-regulated genes were highly enriched with cell cycle-related genes, in addition to genes related to stem cells and apoptosis. Conclusions: Our findings suggest that TRα-mediated T3 activation of the cell cycle program is involved in larval epithelial cell death and adult epithelial stem cell development during intestinal remodeling.

    DOI: 10.1089/thy.2020.0022

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  • The development of adult intestinal stem cells: Insights from studies on thyroid hormone-dependent anuran metamorphosis. 査読 国際誌

    Yun-Bo Shi, Yuki Shibata, Yuta Tanizaki, Liezhen Fu

    Vitamins and hormones   116   269 - 293   2021年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Vertebrates organ development often takes place in two phases: initial formation and subsequent maturation into the adult form. This is exemplified by the intestine. In mouse, the intestine at birth has villus, where most differentiated epithelial cells are located, but lacks any crypts, where adult intestinal stem cells reside. The crypt is formed during the first 3 weeks after birth when plasma thyroid hormone (T3) levels are high. Similarly, in anurans, the intestine undergoes drastic remodeling into the adult form during metamorphosis in a process completely dependent on T3. Studies on Xenopus metamorphosis have revealed important clues on the formation of the adult intestine during metamorphosis. Here we will review our current understanding on how T3 induces the degeneration of larval epithelium and de novo formation of adult intestinal stem cells. We will also discuss the mechanistic conservations in intestinal development between anurans and mammals.

    DOI: 10.1016/bs.vh.2021.02.010

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  • Thyroid hormone receptor beta is critical for intestinal remodeling during Xenopus tropicalis metamorphosis 査読

    Yuki Shibata, Yuta Tanizaki, Yun-Bo Shi

    Cell & Bioscience   10 ( 1 )   2020年12月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer Science and Business Media LLC  

    <title>Abstract</title><sec>
    <title>Background</title>
    Thyroid hormone (T3) is critical for development in all vertebrates. The mechanism underlying T3 effect has been difficult to study due to the uterus-enclosed nature of mammalian embryos. Anuran metamorphosis, which is dependent on T3 but independent of maternal influence, is an excellent model to study the roles of T3 and its receptors (TRs) during vertebrate development. We and others have reported various effects of <italic>TR</italic> knockout (<italic>TRα</italic> and <italic>TRβ</italic>) during <italic>Xenopus tropicalis</italic> development. However, these studies were largely focused on external morphology.


    </sec><sec>
    <title>Results</title>
    We have generated <italic>TRβ</italic> knockout animals containing an out-frame-mutation of 5 base deletion by using the CRISPR/Cas9 system and observed that <italic>TRβ</italic> knockout does not affect premetamorphic tadpole development. We have found that the basal expression of direct T3-inducible genes is increased but their upregulation by T3 is reduced in the intestine of premetamorphic homozygous <italic>TRβ</italic> knockout animals, accompanied by reduced target binding by TR. More importantly, we have observed reduced adult stem cell proliferation and larval epithelial apoptosis in the intestine during T3-induced metamorphosis.


    </sec><sec>
    <title>Conclusions</title>
    Our data suggest that <italic>TRβ</italic> plays a critical role in intestinal remodeling during metamorphosis.


    </sec>

    DOI: 10.1186/s13578-020-00411-5

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    その他リンク: http://link.springer.com/article/10.1186/s13578-020-00411-5/fulltext.html

  • Knocking out histone methyltransferase PRMT1 leads to stalled tadpole development and lethality in Xenopus tropicalis. 査読 国際誌

    Yuki Shibata, Morihiro Okada, Thomas C Miller, Yun-Bo Shi

    Biochimica et biophysica acta. General subjects   1864 ( 3 )   129482 - 129482   2020年3月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Asymmetric arginine dimethylation of histone H4R3 to H4R3me2a by protein arginine methyltransferase 1 (PRMT1) has been implicated to play a key role in gene activation throughout vertebrates. PRMT1 knockout in mouse leads to embryonic lethality. This and the uterus-enclosed nature of the mouse embryo make it difficult to determine the development role of PRMT1 in mammals. METHODS: We took advantage of the external development of the diploid anuran Xenopus tropicalis and adapted the TALEN genome editing technology to knock out PRMT1 in order to investigate how PRMT1 participates in vertebrate development. RESULTS: We observed that PRMT1 knockout had no apparent effect on embryogenesis because normally feeding tadpoles were formed, despite the reduced asymmetric H4R3 di-methylation (H4R3me2a) due to the knockout. However, PRMT1 knockout tadpoles had severely reduced growth even with normal growth hormone gene expression. These tadpoles were also stalled in development shortly after feeding began at stages 44/45 and died within 2 weeks, well before the onset of metamorphosis. In situ analyses revealed broad cessation or drastic reduction in cell proliferation in diverse organs including the eye, brain, spinal cord, liver, and intestine. CONCLUSIONS: Our findings suggest that PRMT1 is not required for embryogenesis but is a key regulator for normal progression of vertebrate development and growth. GENERAL SIGNIFICANCE: The similarities and differences between PRMT1 knockout Xenopus tropicalis and mouse suggest that two distinct phases of vertebrate development: early embryogenesis and subsequent growth/organ maturation, have different but evolutionally conserved requirement for epigenetic modifications.

    DOI: 10.1016/j.bbagen.2019.129482

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  • Functional Studies of Transcriptional Cofactors via Microinjection-Mediated Gene Editing in Xenopus. 査読 国際誌

    Yuki Shibata, Lingyu Bao, Liezhen Fu, Bingyin Shi, Yun-Bo Shi

    Methods in molecular biology (Clifton, N.J.)   1874   507 - 524   2019年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The anuran Xenopus laevis has been studied for decades as a model for vertebrate cell and developmental biology. More recently, the highly related species Xenopus tropicalis has offered the opportunity to carry out genetic studies due to its diploid genome as compared to the pseudo-tetraploid Xenopus laevis. Amphibians undergo a biphasic development: embryogenesis to produce a free-living tadpoles and subsequent metamorphosis to transform the tadpole to a frog. This second phase mimics the so-called postembryonic development in mammals when many organs/tissues mature into their adult form in the presence of high levels of plasma thyroid hormone (T3). The total dependence of amphibian metamorphosis on T3 offers a unique opportunity to study postembryonic development in vertebrates, especially with the recent development gene editing technologies that function in amphibians. Here, we first review the basic molecular understanding of the regulation of Xenopus metamorphosis by T3 and T3 receptors (TRs), and then describe a detailed method to use CRISPR to knock out the TR-coactivator SRC3 (steroid receptor coactivator 3), a histone acetyltransferase, in order to study its involvement in gene regulation by T3 in vivo and Xenopus development.

    DOI: 10.1007/978-1-4939-8831-0_29

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  • Thyroid Hormone Receptor α Controls Developmental Timing and Regulates the Rate and Coordination of Tissue-Specific Metamorphosis in Xenopus tropicalis. 査読 国際誌

    Luan Wen, Yuki Shibata, Dan Su, Liezhen Fu, Nga Luu, Yun-Bo Shi

    Endocrinology   158 ( 6 )   1985 - 1998   2017年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Thyroid hormone (T3) receptors (TRs) mediate the effects of T3 on organ metabolism and animal development. There are two TR genes, TRα and TRβ, in all vertebrates. During animal development, TRα expression is activated earlier than zygotic T3 synthesis and secretion into the plasma, implicating a developmental role of TRα both in the presence and absence of T3. Using T3-dependent amphibian metamorphosis as a model, we previously proposed a dual-function model for TRs, in particular TRα, during development. That is, unliganded TR represses the expression of T3-inducible genes during premetamorphosis to ensure proper animal growth and prevent premature metamorphosis, whereas during metamorphosis, liganded TR activates target gene transcription to promote the transformation of the tadpole into a frog. To determine if TRα has such a dual function, we generated homozygous TRα-knockout animal lines. We show that, indeed, TRα knockout affects both premetamorphic animal development and metamorphosis. Surprisingly, we observed that TRα is not essential for amphibian metamorphosis, given that homozygous knockout animals complete metamorphosis within a similar time period after fertilization as their wild-type siblings. On the other hand, the timing of metamorphosis for different organs is altered by the knockout; limb metamorphosis occurs earlier, whereas intestinal metamorphosis is completed later than in wild-type siblings. Thus, our studies have demonstrated a critical role of endogenous TRα, not only in regulating both the timing and rate of metamorphosis, but also in coordinating temporal metamorphosis of different organs.

    DOI: 10.1210/en.2016-1953

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  • Molecular and cellular characterization of urinary bladder-type aquaporin in Xenopus laevis. 査読 国際誌

    Yuki Shibata, Izumi Katayama, Takashi Nakakura, Yuji Ogushi, Reiko Okada, Shigeyasu Tanaka, Masakazu Suzuki

    General and comparative endocrinology   222   11 - 9   2015年10月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In contrast to many anuran amphibians, water is not reabsorbed from the urinary bladder in aquatic Xenopus, thereby helping to prevent excessive water influx. However, little is known about the molecular mechanisms for this process. In the present study, we have identified urinary bladder-type aquaporin, AQP-x2, in Xenopus laevis by cDNA cloning. The predicted amino acid sequence contained six putative transmembrane domains and the two conserved Asn-Pro-Ala motifs, characteristic of AQPs. The sequence also contained a putative N-glycosylation site and phosphorylation motifs for protein kinase A and protein kinase C. The oocyte swelling assay showed that AQP-x2 facilitated water permeability. Reverse transcription-PCR analysis indicated that AQP-x2 mRNA was expressed in the urinary bladder and lung, and faintly in the kidney. Immunomicroscopical study further localized AQP-x2 protein to the cytoplasm of granular cells in the luminal epithelium of the urinary bladder whilst AQP3 was observed along the basolateral side of these cells. In vitro stimulation of the urinary bladder with 10(-8)M vasotocin (AVT), 10(-8)M hydrin 1, or 10(-8)M hydrin 2 had no clear effect on the subcellular distribution of AQP-x2. When the AVT concentration was increased to 10(-6)M, however, AQP-x2 was partially transferred to the apical plasma membrane. The treatment with hydrin 1 or hydrin 2 at the same concentration failed to induce the translocation to the apical membrane. On the other hand, AQP3 remained along the basolateral side even after the treatment with vasotocin or hydrins. The results suggest that the poor responsiveness of AQP-x2 to neurohypophyseal peptides may be a main cause for the little water permeability of the urinary bladder of X. laevis.

    DOI: 10.1016/j.ygcen.2014.09.001

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  • Molecular machinery for vasotocin-dependent transepithelial water movement in amphibians: aquaporins and evolution. 査読 国際誌

    Masakazu Suzuki, Yuki Shibata, Yuji Ogushi, Reiko Okada

    The Biological bulletin   229 ( 1 )   109 - 19   2015年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Amphibians represent the first vertebrates to adapt to terrestrial environments, and are successfully distributed around the world. The ventral skin, kidney, and urinary bladder are important osmoregulatory organs for adult anuran amphibians. Water channel proteins, called aquaporins (AQPs), play key roles in transepithelial water absorption/reabsorption in these organs. At least 43 types of AQPs were identified in anurans; a recent phylogenetic analysis categorized anuran AQPs among 16 classes (AQP0-14, 16). Anuran-specific AQPa2 was assigned to AQP6, then was further subdivided into the ventral skin-type (AQP6vs; AQPa2S), whose expression is confined to the ventral skin, and the urinary bladder-type (AQP6ub; AQPa2U), which is basically expressed in the urinary bladder. For the osmoregulatory organs, AQP3 is constitutively located in the basolateral plasma membrane of tight-junctioned epithelial cells. AQP6vs, AQP2 and/or AQP6ub are also expressed in these epithelial cells and are translocated to the apical membrane in response to arginine vasotocin, thereby regulating water absorption/reabsorption. It was suggested recently that two subtypes of AQP6vs contribute to cutaneous water absorption in Ranid species. In addition, AQP5 (AQP5a) and AQP5L (AQP5b) were identified from Xenopus tropicalis Gray, 1864, and AQP5 was localized to the apical membrane of luminal epithelial cells of the urinary bladder in dehydrated Xenopus. This finding suggested that AQP5 may be involved in water reabsorption from this organ under dehydration. Based on the hitherto reported information, we propose models for the evolution of water-absorbing/reabsorbing mechanisms in anuran osmoregulatory organs in association with AQPs.

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  • Aquaporins and water homeostasis in Xenopus

    Shibata Y, Suzuki M

    Advances in Animal Science and zoology   7   119 - 134   2015年

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    担当区分:筆頭著者  

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  • Gene expression and localization of two types of AQP5 in Xenopus tropicalis under hydration and dehydration. 査読 国際誌

    Yuki Shibata, Takahiro Sano, Nobuhito Tsuchiya, Reiko Okada, Hiroshi Mochida, Shigeyasu Tanaka, Masakazu Suzuki

    American journal of physiology. Regulatory, integrative and comparative physiology   307 ( 1 )   R44-56   2014年7月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Two types of aquaporin 5 (AQP5) genes (aqp-xt5a and aqp-xt5b) were identified in the genome of Xenopus tropicalis by synteny comparison and molecular phylogenetic analysis. When the frogs were in water, AQP-xt5a mRNA was expressed in the skin and urinary bladder. The expression of AQP-xt5a mRNA was significantly increased in dehydrated frogs. AQP-xt5b mRNA was also detected in the skin and increased in response to dehydration. Additionally, AQP-xt5b mRNA began to be slightly expressed in the lung and stomach after dehydration. For the pelvic skin of hydrated frogs, immunofluorescence staining localized AQP-xt5a and AQP-xt5b to the cytoplasm of secretory cells of the granular glands and the apical plasma membrane of secretory cells of the small granular glands, respectively. After dehydration, the locations of both AQPs in their respective glands did not change, but AQP-xt5a was visualized in the cytoplasm of secretory cells of the small granular glands. For the urinary bladder, AQP-xt5a was observed in the apical plasma membrane and cytoplasm of a number of granular cells under normal hydration. After dehydration, AQP-xt5a was found in the apical membrane and cytoplasm of most granular cells. Injection of vasotocin into hydrated frogs did not induce these changes in the localization of AQP-xt5a in the small granular glands and urinary bladder, however. The results suggest that AQP-xt5a might be involved in water reabsorption from the urinary bladder during dehydration, whereas AQP-xt5b might play a role in water secretion from the small granular gland.

    DOI: 10.1152/ajpregu.00186.2013

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  • Novel vasotocin-regulated aquaporins expressed in the ventral skin of semiaquatic anuran amphibians: evolution of cutaneous water-absorbing mechanisms. 査読 国際誌

    Yasunori Saitoh, Yuji Ogushi, Yuki Shibata, Reiko Okada, Shigeyasu Tanaka, Masakazu Suzuki

    Endocrinology   155 ( 6 )   2166 - 77   2014年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Until now, it was believed that only one form of arginine vasotocin (AVT)-regulated aquaporin (AQP) existed to control water absorption from the ventral skin of semiaquatic anuran amphibians, eg, AQP-rj3(a) in Rana japonica. In the present study, we have identified a novel form of ventral skin-type AQP, AQP-rj3b, in R. japonica by cDNA cloning. The oocyte swelling assay confirmed that AQP-rj3b can facilitate water permeability. Both AQP-rj3a and AQP-rj3b were expressed abundantly in the ventral hindlimb skin and weakly in the ventral pelvic skin. For the hindlimb skin, water permeability was increased in response to AVT, although the hydroosmotic response was not statistically significant in the pelvic skin. Isoproterenol augmented water permeability of the hindlimb skin, and the response was inhibited by propranolol. These events were well correlated with the intracellular trafficking of the AQPs. Immunohistochemistry showed that both AQP-rj3 proteins were translocated from the cytoplasmic pool to the apical membrane of principal cells in the first-reacting cell layer of the hindlimb skin after stimulation with AVT and/or isoproterenol. The type-b AQP was also found in R. (Lithobates) catesbeiana and R. (Pelophylax) nigromaculata. Molecular phylogenetic analysis indicated that the type-a is closely related to ventral skin-type AQPs from aquatic Xenopus, whereas the type-b is closer to the AQPs from terrestrial Bufo and Hyla, suggesting that the AQPs from terrestrial species are not the orthologue of the AQPs from aquatic species. Based on these results, we propose a model for the evolution of cutaneous water-absorbing mechanisms in association with AQPs.

    DOI: 10.1210/en.2013-1928

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  • Localization of water channels in the skin of two species of desert toads, Anaxyrus (Bufo) punctatus and Incilius (Bufo) alvarius. 査読

    Yuki Shibata, Hiro-Aki Takeuchi, Takahiro Hasegawa, Masakazu Suzuki, Shigeyasu Tanaka, Stanley D Hillyard, Takatoshi Nagai

    Zoological science   28 ( 9 )   664 - 70   2011年9月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Anuran amphibians obtain water by osmosis across their ventral skin. A specialized region in the pelvic skin of semiterrestrial species, termed the seat patch, contains aquaporins (AQPs) that become inserted into the apical plasma membrane of the epidermis following stimulation by arginine vasotocin (AVT) to facilitate rehydration. Two AVT-stimulated AQPs, AQP-h2 and AQP-h3, have been identified in the epidermis of seat patch skin of the Japanese tree frog, Hyla japonica, and show a high degree of homology with those of bufonid species. We used antibodies raised against AQP-h2 and AQP-h3 to characterize the expression of homologous AQPs in the skin of two species of toads that inhabit arid desert regions of southwestern North America. Western blot analysis of proteins gave positive results for AQP-h2-like proteins in the pelvic skin and also the urinary bladder of Anaxyrus (Bufo) punctatus while AQP-h3-like proteins were found in extracts from the pelvic skin and the more anterior ventral skin, but not the urinary bladder. Immunohistochemical observations showed both AQP-h2- and AQP-h3-like proteins were present in the apical membrane of skin from the pelvic skin of hydrated and dehydrated A. punctatus. Further stimulation by AVT or isoproterenol treatment of living toads was not evident. In contrast, skin from hydrated Incilius (Bufo) alvarius showed very weak labeling of AQP-h2- and AQP-h3-like proteins and labeling turned intense following stimulation by AVT. These results are similar to those of tree frogs and toads that occupy mesic habitats and suggest this pattern of AQP expression is the result of phylogenetic factors shared by hylid and bufonid anurans.

    DOI: 10.2108/zsj.28.664

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▼全件表示

書籍等出版物

  • Aquaporins and water homeostasis in Xenopus

    Shibata Y, Suzuki M( 担当: 共著 範囲: Advances in Animal Science and zoology, 7: 119-134)

    Nova Science Publishers  2015年 

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講演・口頭発表等

  • アフリカツメガエルの新たなトランスジェニック動物作製法 〜NEXTrans (New and Easy Xenopus Transgenesis at a safe harbor site)〜 招待

    柴田侑毅, 奥村晃成, 餅井真, 鈴木賢一

    第93回日本動物学会 シンポジウムS12「新規モデル生物で遺伝子ノックイン技術を駆使する研究の醍醐味〜先人の試行錯誤から成功の秘訣を学ぶ」  2022年9月 

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    開催年月日: 2022年9月

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  • ネッタイツメガエル(Xenopus tropicalis)における AQP6/a2S の発現制御に関する分子生物 学的研究

    沼田理穏, 柴田侑毅, 小竹康平, 岡田令子, 鈴木雅一

    第45回日本比較内分泌学会  2021年11月 

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    開催年月日: 2021年11月

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  • ネッタイツメガエルの変態における甲状腺ホルモン受容体の役割 招待

    柴田侑毅, Yun-Bo Shi

    第92回日本動物学会  2021年9月 

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    開催年月日: 2021年9月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

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  • The function of the endogenous protein arginine methyltransferase 1 (PRMT1) gene during post-embryonic development in Xenopus tropicalis.

    Shibata Y, Okada M, Shi YB

    American thyroid association annual meeting  2018年9月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • Thyroid hormone receptor is indispensable for metamorphosis in Xenopus tropicalis

    Shibata Y, Okada M, Shi YB

    NICHD DIR & DIPHR Joint Scientific Retreat  2018年9月 

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    記述言語:英語   会議種別:ポスター発表  

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  • Molecular function of thyroid hormone receptor during frog development

    Shibata Y, Shi YB

    43th Annual meeting of Japan society of comparative endocrinology  2019年 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • Tissue-specific roles of thyroid hormone receptors ensures coordinated and complete metamorphosis in Xenopus tropicalis

    Shibata Y, Shi YB

    90th Annual meeting of the zoological society of japan  2019年 

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    記述言語:日本語   会議種別:口頭発表(一般)  

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  • THYROID HORMONE RECEPTOR DEFFICIENCY ACCELERATES CONNECTIVE TISSUE DEVELOPMENT BUT PREVENTS EPITHELIAL TRANSFORMATION DURING METAMORPHOSIS IN XENOPUS TROPICALIS.

    Shibata Y, Okada M, Shi YB

    5th Biennial meeting, North American Society of Comparative Endocrinology (NASCE)  2019年 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • カエルツボカビ(Batrachochytriumdendrobatidis)感染が無尾両生類の腹側皮膚を介した水 吸収機構に与える影響

    青島泰雅, 柴田侑毅, 嘉手苅将, 石井 凌, 常盤俊大, 宇根有美, 鈴木雅一

    第45回日本比較内分泌学会  2021年11月 

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  • CRISPR/Cas9を用いた新規トランスジェニックカエル作製法の開発

    柴田侑毅, 餅井 真, 鈴木賢一

    第45回日本比較内分泌学会  2021年11月 

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  • Molecular evolution and physiological roles of aquaporins expressed in anuran osmoregulatory organs 招待

    Masakazu Suzuki, Yuki Shibata, Reiko Okada

    The 8th International Symposium on Amphibian and Reptilian Endocrinology and Neurobiology 

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    記述言語:英語  

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  • 乾燥状態におけるネッタイツメガエルに発現する新規アクアポリンAQP-xt5’

    柴田侑毅, 原純也, 岡田令子, 鈴木雅一, 田中滋康

    第81回 日本動物学会大会  2010年9月 

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    記述言語:日本語  

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  • ネッタイツメガエルXenopus tropicalis夏眠時における窒素排出機構の変化

    宮崎翼, 佐野貴太, 柴田侑毅, 鈴木雅一, 岡田令子

    第82回日本動物学会大会  2011年9月 

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  • 乾燥状態におけるネッタイツメガエルの水恒常性維持に関する研究

    柴田侑毅, 佐野貴太, 滝田優, 鈴木雅一, 田中滋康

    第35回日本比較内分泌学会大会  2010年11月 

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    記述言語:日本語   会議種別:ポスター発表  

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  • ネッタイツメガエル(Xenopus tropicalis)の環境適応におけるアクアポリン発現解析

    柴田侑毅, 佐野貴太, 原純也, 鈴木雅一, 田中滋康

    第82回日本動物学会大会  2011年9月 

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  • ネッタイツメガエルの乾燥適応と尿素回路の変化

    宮崎翼, 柴田侑毅, 佐野貴太, 鈴木雅一, 岡田令子

    第37回日本比較内分泌学会大会  2012年11月 

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  • 両生類特異的なAVT調節性アクアポリン(AQP)の分子進化

    柴田侑毅 , SD Hillyard , 鈴木雅一 , 田中滋康 , 長井孝紀

    第37回日本比較内分泌学会大会  2012年11月 

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  • ネッタイツメガエルXenopus tropicalisを用いた環境変化と水恒常性維持機構に関する研究

    柴田侑毅, 宮崎翼, 佐野貴太, 鈴木雅一, 田中滋康

    第36回日本比較内分泌学会大会  2011年11月 

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  • 極限水環境におけるネッタイツメガエル(Xenopus tropicalis)のアクアポリン発現解析

    柴田侑毅 , 宮崎翼 , 佐野貴太 , 岡田令子 , 鈴木雅一

    第83回日本動物学会大会  2012年9月 

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  • 二ホンアマガエルの変態過程におけるアクアポリンの発現解析

    山下詩織 , 佐野晴奈 , 柴田侑毅 , 岡田令子 , 鈴木雅一

    第84回日本動物学会大会  2013年9月 

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  • 有尾両生類に発現する両生類特異的なAVT 調節性アクアポリン(AQP)の機能と分子進化

    柴田侑毅 , S.D. Hillyard , 田中滋康 , 鈴木 雅一 , 長井孝紀

    平成25 年度日本動物学会中部支部大会  2014年3月 

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  • Molecular evolution and function of the orthologue of mammalian aquaporin 5 in anuran and urodele amphibians

    Y. Shibata, SD Hillyard, M. Suzuki, T. Nagai, S. Tanaka

    17th International Congress of Comparative Endocrinology  2013年7月 

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  • ネッタイツメガエルの腹側皮膚型アクアポリン(AQP)を介した極限水環境への順応機構

    柴田侑毅, 佐野貴太, 岡田令子, 鈴木雅一, 田中滋康

    第84回日本動物学会大会  2013年9月 

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  • 有尾両生類における腹側皮膚型アクアポリン(AQPa2S)の機能と分子進化

    柴田侑毅 , Stanley D Hillyard , 岡田令子 , 鈴木雅一 , 長井孝紀

    第85回日本動物学会大会  2014年9月 

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  • Influence of the chytrid fungus on cutaneous water permeability of anuran amphibians

    Yuki Shibata, Sho Kadekaru, Toshihiro Tokiwa, Yumi Une, Masakazu Suzuki

    ELSU symposium 2014  2014年9月 

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  • Anuran-specific aquaporin 2 ortholog in a urodele, evidence for early gene duplication in amphibian evolution

    Yuki Shibata , Masakazu Suzuki , Takatoshi Nagai , Stanley Hillyard

    APS fall meeting  2014年10月 

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  • Spatial and temporal expression of aquaporins, AQP2 and AQP3, in the kidney during metamorphosis of the tree frog, Hyla japonica

    Takayuki Ueno, Yuki Shibata, Haruna Sano, Shiori Yamashita, Masakazu Suzuki

    The 8th International Symposium on Amphibian and Reptilian Endocrinology and Neurobiology  2014年11月 

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  • Anuran-like aquaporins in the skin and urinary bladder of a urodele, evidence for early gene duplication in the lissamphibia

    Stanley D. Hillyard, Yuki Shibata, Takatoshi Nagai, Masakazu Suzuki

    Experimental Biology 2015  2015年3月 

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  • Expression and function of aquaporins in Xenopus tropicalis under hydration and dehydration

    第39回日本比較内分泌学会大会  2014年11月 

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  • カエルツボカビ(Batrachochytrium dendrobatidis)が無尾両生類の皮膚を介した水透過性に与える影響

    柴田侑毅, 嘉手苅将, 常盤俊大, 宇根有美, 鈴木雅一

    平成26年度 日本動物学会中部支部大会能登大会  2014年11月 

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  • Effect on cutaneous absorption of the green tree frog by Batrachochytrium dendrobatidis infection

    Kadekaru S, Shibata S, Tokiwa T, Suzuki M, Une Y

    American college of veterinary pathologists  2017年11月 

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    記述言語:英語   会議種別:ポスター発表  

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  • Function of the histone methyltransferase PRMT1 during post-embryonic development and adult intestinal stem cell formation in Xenopus tropicalis.

    Shibata Y, Okada M, Shi YB

    American thyroid association anural meeting  2017年11月 

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    記述言語:英語   会議種別:ポスター発表  

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  • カエルツボカビ感染が皮膚水吸収に与える影響

    嘉手苅 将, 柴田 侑毅, 常盤 俊大, 鈴木 雅一, 宇根 有美

    第3回獣医病理学専門家協会学術集会  2016年3月 

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  • Further characterization of anuran-like aquaporins in the urinary bladder of urodeles, Notopthalmus viridescens and Cynops pyrrhogaster.

    Hillyard S, Shibata Y, Ishii R, Nagai T, Suzuki M

    The Experimental Biology meeting  2016年4月 

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    記述言語:英語   会議種別:ポスター発表  

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  • Investigating the function of the histone methyltransferase PRMT1 during post-embryonic development in Xenopus tropicalis

    Shibata Y, Okada M, Shi YB

    The Experimental Biology  2018年4月 

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    記述言語:英語   会議種別:ポスター発表  

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  • Genetic analysis of thyroid hormone regulation of intestinal remodeling in Xenopus 招待

    Yuki Shibata, Kenta Fujimoto, Ken-ichi T Suzuki, Takashi Hasebe

    7th Biennial Meeting of The North American Society for Comparative Endocrinology  2023年5月 

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  • NEXTrans: Simple transgenesis at a novel safe harbor site by using CRISPR-Cas9 in Xenopus laevis

    Yuki Shibata, Akinori Okumura, Makoto Mochii, Ken-ichi T. Suzuki

    19th International Xenopus Conference  2023年8月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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▼全件表示

受賞

  • NIBB若手研究者賞

    2021年12月  

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  • Ming K. Jeang Award for Excellence in Cell & Bioscience:

    2021年10月   Cell and Bioscience (BMC)   Thyroid hormone receptor beta is critical for intestinal remodeling during Xenopus tropicalis metamorphosis:

    Yuki Shibata, Yuta Tanizaki, Yun-Bo Shi

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  • Fellows Award for Research Excellence (FARE)

    2020年   National Institute of Health (NIH)  

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  • NASCE Student Hotel Award

    2019年5月   North American Society for Comparative Endocrinology (NASCE)  

    Yuki Shibata

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  • Trainee Grant Program (Travel award)

    2018年   American Thyroid Association  

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  • 若手最優秀発表賞

    2018年   日本比較内分泌学会  

    柴田侑毅

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  • Trainee Grant Program (Travel award)

    2017年   American Thyroid Association  

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  • 優秀発表賞(博士後期課程ポスター発表の部)

    2014年   日本動物学会 中部支部大会  

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共同研究・競争的資金等の研究課題

  • 空間トランスクリプトームによる消化管上皮幹細胞形成に関わる遺伝子の探索と機能解析

    研究課題/領域番号:23K05834  2023年4月 - 2026年3月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    柴田 侑毅

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    配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

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  • Cas13dを用いた両生類の四肢発生に関わる遺伝子の時期特異的ノックダウン

    研究課題/領域番号:21K20667  2021年8月 - 2023年3月

    日本学術振興会  科学研究費助成事業 研究活動スタート支援  研究活動スタート支援

    柴田 侑毅

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    配分額:3120000円 ( 直接経費:2400000円 、 間接経費:720000円 )

    研究課題であるCas13dとTetOnシステムを組み合わせた系を確立するため、本年度はまずヒートショックプロモーターの下流でTetOn3Gタンパクを発現するトランスジェニック(Tg)カエル(Tg(hsp:teton3g:cry:tdtomato))の作製に取り組んだ。この際、CRISPR/Cas9を用いて、ゲノムターゲティングを基とした新規Tgアフリカツメガエルの作製法(NEXTrans)を開発した。NEXTrans法は、ホストゲノムの特定領域が組み込まれたベクター、特定領域内で設計されたsgRNAおよびCas9タンパク質をインジェクションすることで、高効率で特定の領域に標的DNAを組み込んだTgを簡便に作製できる(ゲノムターゲティング)。本手法の開発にあたり、Fin and gill keratin (FGK) promoter、Crysterin promoter、CMV promoter、Sox2 promoterの下流にGFPやtdTomatoを組み込み、組織特異的に蛍光タンパク質が発現することを確認している。しかしながら、組み込まれた外来遺伝子が次世代に継承するgermline transmissionを確認するに至っていないため、性成熟の完了を待ち、早急に解析を進める予定である。本手法は広くTg作製に用いられているREMI法やI-SceI法に加え、第3のTg作製オプションとなることが期待される。NEXTrans法を用いて、今年度の目標であったTg(hsp:teton3g:cry:tdtomato)個体のF0世代を得ることに成功した。

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  • Investigating the function of the histone methyltransferase PRMT1 during thyroid hormone dependent metamorphosis and adult intestinal stem cell development in Xenopus tropicalis

    2017年3月 - 2019年2月

    JSPS Research Fellowship for Japanese Biomedical and Behavioral Researchers at NIH (海特NIH) 

    柴田侑毅

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  • 両生類におけるアクアポリンの生理学的役割、遺伝子発現調節機構、および進化プロセス

    研究課題/領域番号:13J04065  2013年4月 - 2015年3月

    日本学術振興会  科学研究費助成事業 特別研究員奨励費 (DC2)  特別研究員奨励費

    柴田 侑毅

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    配分額:2000000円 ( 直接経費:2000000円 )

    減水環境で飼育したネッタイツメガエルは、水中環境では示さない腹側皮膚を介した水吸収を行うようになり、乾燥した環境に順応する可能性があることを1年目の研究で示した。さらに、減水環境で飼育するとAQPa2S mRNAおよびタンパク質の発現が確認された。そこで水棲無尾両生類の乾燥環境適応に最も重要であると考えられるAQPa2Sの転写調節機構の解明を目的として以下の実験を行った。減水環境または水中で飼育したネッタイツメガエルの大腿部皮膚をcollagenase 処理し、表皮と真皮を分離した。このうち表皮のみを液体窒素で凍結し、次世代シーケンサーMiseq (illumina)を用いたトランスクリプトーム解析を行った。その結果、減水環境において発現量が増加した203個の因子および発現量が低下した424個の因子を同定した。今後AQPa2Sの転写制御に関わる因子の同定が期待される。
    共同研究者であるHillyard教授の研究室において陸棲の有尾両生類ブチイモリを用いて、腹側皮膚を介した水吸収機構の生理学的な解析を行った。その結果、他の水棲(アカハライモリ)・半水棲(シリケンイモリ)のイモリに対して、ブチイモリのみがホルモン投与に応答して、腹側皮膚を介した水吸収を行うこと示した。また、免疫組織学的な解析からAQPa2SはAVTに応答して最外顆粒細胞のアピカル側細胞膜に局在することを示した。これらの結果は、陸棲のブチイモリが多くの陸棲や樹上棲の無尾両生類と同様に、抗利尿ホルモン応答性AQPa2Sの局在を変化させることで、腹側皮膚から水を吸収して水恒常性を維持していることを示している。本研究により、有尾両生類が無尾両生類と同様の腹側皮膚型AQP(AQPa2S)を介した水吸収機構を有していることが示された。これは同時に、本水吸収機構の起源が無尾目と有尾目の共通の祖先にまで遡ることが出来ることを示している。

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