哺乳動物網膜をモデル系としたエピジェネティックな制御による細胞運命決定機構の研究

• Project Area: Molecular mechanisms of cell fate determination in the cells that undergo stepwise differentiation to multiple pathways
• Project/Area Number: 25118708
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 渡邉 すみ子 東京大学, 医科学研究所, 特任教授 (60240735)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥11,180,000 (Direct Cost: ¥8,600,000、Indirect Cost: ¥2,580,000); Fiscal Year 2014: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2013: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
• Keywords: 網膜 / ヒストンメチル化 / 分化 / 成熟 / 双極細胞 / 運命決定

Outline of Annual Research Achievements

初年度にEzh2, Jmjd3の網膜発生、特に双極細胞の発生における役割があきらかになったので、この分子メカニズムを明らかにするため、会合分子のスクリーニングを行った。タグ付きのEzh2, Jmjd3を作製し、網膜に発現させ、免疫沈降法による共沈実験をおこなった。また、発現パターンの検討などにより、Jmjd3と会合する可能性のある候補の転写因子がひとつあがり、これについて、過剰発現を単独、Jmjd3との二重過剰発現などをおこない、網膜細胞分化への影響を免疫染色により検討した。その結果、この転写因子がJmjd3を標的遺伝子座に誘導してH3K27トリメチル化にいたる機構が想定された。一方、H3K4トリメチル化の役割をH3K4methyltransferaseのsh-RNAの網膜培養系における発現などで検討した。十数種類のメチル化酵素が存在するためこれらすべてに対しsh-RNAを作製し、順次、あるいは組み合わせにより網膜培養系に発現させ、その増殖、分化への影響を凍結切片の免疫染色により観察した。その結果、双極細胞関連遺伝子については、H3K4トリメチル化はその発現制御について重要な役割を果たしていない、という結論になった。一方で視細胞関連遺伝子には発現パターンとH3K4トリメチル化が強く相関するため、細胞系列特異的な役割が想定された。しかしメチル化酵素の数がおおく、特定のメチル化酵素の役割を同定するにはいたらなかった。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Gene activated-matrix comprised of atelocollagen and plasmid DNA encoding Bmp4 or Runx2 promotes rat cranial bone augmentation (2015)
  • Author(s): Umebayashi M., Sumita, Y., Kawai, Y., Watanabe, S., Asahina, I.
  • Journal Title: Bio Research Open Access Volume: -
  • Peer Reviewed / Open Access
• [Journal Article] Ablation of Kcnj10 expression in retinal explants revealed pivotal roles for Kcnj10 in the proliferation and development of Muller glia (2015)
  • Author(s): Arai, E., Baba, Y., Iwagawa, T.,Kuribayashi, H., Mochizuki, Y., Watanabe, S.
  • Journal Title: Mol Vis Volume: 21 Pages: 148-159
  • Peer Reviewed
• [Journal Article] Roles Histone H3K27 tri-methylase Ezh2 in retinal proliferation and differentiation (2015)
  • Author(s): ida, A., Iwagawa, T., Baba, Y.,Satoh, S., Mochizuki, Y., Nakauchi, H., Furukawa, T., Koseki, H., Murakami, A., Watanabe, S.
  • Journal Title: Developmental Neurobiology Volume: - Issue: 9 Pages: 3751-3756
  • DOI: 10.1002/dneu.22261
  • Peer Reviewed
• [Journal Article] DNA methylation is involved in the expression of miR-142-3p in fibroblasts and induced pluripotent stem cells (2014)
  • Author(s): Abdul Razak, S. A., Baba, Y., Nakauchi, H., Otsu, M., Watanabe, S.
  • Journal Title: Stem Cells International Volume: 101349 Pages: 1-8
  • DOI: 10.1155/2014/101349
  • Peer Reviewed
• [Journal Article] Critical Roles of DNase1l3l in Lens Nuclear Degeneration in Zebrafish (2014)
  • Author(s): Iida, A., Tabata, Y., Baba, Y., Fujii, T., Watanabe, S.
  • Journal Title: Biochimie Volume: 106 Pages: 68-74
  • DOI: 10.1016/j.biochi.2014.07.025
  • Peer Reviewed
• [Journal Article] Histone demethylase Jmjd3 is required for the development of subsets of retinal bipolar cells. (2014)
  • Author(s): Iida A, Iwagawa T, Kuribayashi H, Satoh S, Mochizuki Y, Baba Y, Nakauchi H, Furukawa T, Koseki H, Murakami A, Watanabe S.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 111 Issue: 10 Pages: 3751-6
  • DOI: 10.1073/pnas.1311480111
  • Peer Reviewed / Open Access
• [Journal Article] BMP signaling participates in late phase differentiation of the retina, party via upregulation of Hey2 (2014)
  • Author(s): Kuribayashi, H., Baba, Y., Watanabe, S.
  • Journal Title: Developmental Neurobiology Volume: 74 Issue: 12 Pages: 1172-1183
  • DOI: 10.1002/dneu.22196
  • Peer Reviewed
• [Journal Article] Transposon mutagenesis identifies Foxr2 as a putative oncogene in medulloblastoma (2014)
  • Author(s): Koso, H., Tsuhako, A., Lyons, E., Ward, J. M., Rust, A. G., Adams, D. J., Jenkins, N. A., Copeland, N. G., Watanabe, S.
  • Journal Title: Can Re Volume: 74 Pages: 2351-2361
  • Peer Reviewed
• [Journal Article] Use of cell type-specific transcriptome to identify genes specifically involved in Müller glia differentiation during retinal development. (2014)
  • Author(s): Mochizuki Y, Iida A, Lyons E, Kageyama R, Nakauchi H, Murakami A, Watanabe S.
  • Journal Title: Dev Neurobiol. Volume: 74 Issue: 4 Pages: 426-37
  • DOI: 10.1002/dneu.22131
  • Peer Reviewed
• [Journal Article] Transposon mutagenesis identifies Foxr2 as a putative oncogene in medulloblastoma (2014)
  • Author(s): Koso, H. et al.
  • Journal Title: Can Res Volume: 74 Pages: 2351-2361
  • Peer Reviewed
• [Journal Article] The early retinal progenitor-expressed gene Sox11 regulates the timing of the differentiation of retinal cells (2013)
  • Author(s): Usui, A. et al.
  • Journal Title: Development Volume: 140 Issue: 4 Pages: 740-750
  • DOI: 10.1242/dev.090274
  • Peer Reviewed
• [Journal Article] Robust long-term transduction of common marmoset neuromusculartissue with rAAV1 and rAAV9. (2013)
  • Author(s): Hironori Okada, et al.
  • Journal Title: Molecular Therapy-Nucleic Acids Volume: 2 Pages: e95-e95
  • DOI: 10.1038/mtna.2013.21
  • Peer Reviewed
• [Journal Article] Enhancer/promoter activities of the long/middle wavelength-sensitive opsins of vertebrates mediated by thyroid hormone receptor β2 and COUP-TFII (2013)
  • Author(s): Iwagawa, T. et al.
  • Journal Title: Plos One Volume: 8 Issue: 8 Pages: e72065-e72065
  • DOI: 10.1371/journal.pone.0072065
  • Peer Reviewed / Open Access
• [Journal Article] Profiling of MicroRNA in Human and Mouse ES and iPS Cells Reveals Overlapping but Distinct MicroRNA Expression Patterns. (2013)
  • Author(s): Razak SR, Ueno K, Takayama N, Nariai N, Nagasaki M, Saito R, Koso H, Lai CY, Murakami M, Tsuji K, Michiue T, Nakauchi H, Otsu M, Watanabe S.
  • Journal Title: PLoS One. Volume: 8 Issue: 9 Pages: e73532-e73532
  • DOI: 10.1371/journal.pone.0073532
  • Peer Reviewed / Open Access