Analyses of genes involved in cell polarity in tissues and animals in situ

• Project/Area Number: 24390046
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: General anatomy (including Histology/Embryology)
• Research Institution: Osaka University
• Principal Investigator: HARADA Akihiro 大阪大学, 医学(系)研究科(研究院), 教授 (40251441)
• Co-Investigator(Kenkyū-buntansha): YOSHIMURA Shin'ichiro 大阪大学, 大学院医学系研究科, 助教 (60584521) ; KUNII Masataka 大阪大学, 大学院医学系研究科, 助教 (80614768)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥18,720,000 (Direct Cost: ¥14,400,000、Indirect Cost: ¥4,320,000); Fiscal Year 2014: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2012: ¥7,020,000 (Direct Cost: ¥5,400,000、Indirect Cost: ¥1,620,000)
• Keywords: 細胞極性 / 極性輸送 / ノックアウトマウス / apical / 上皮細胞 / 神経細胞

Outline of Final Research Achievements

We published papers containing analyses of Rab8a,b, PKD1,2, and FAPP2 knockout mice. We finished analysis of pancreas-specific SNAP23 knockout mice and identified molecular mechanism for increased insulin secretion in these mice. The results were summarized and submitted to a journal. We are analyzing the molecular mechanisms underlining phenotypes of syntaxin3 knockout mice by generating KO cell lines by CRISPR/Cas9 method. For identifying molecular mechanism of apical transport, we discovered a Rab8a binding protein and its role for apical transport. For monitoring the transport of apical proteins, we generated a model protein and introduced it into cultured intestinal organoids. We found out novel candidate molecules for apical transport using C. elegans. We generated knockout mice lacking these molecules and analyzing their phenotypes.

Research Products

• [Journal Article] The Role of PKD in Cell Polarity, Biosynthetic Pathways, and Organelle/F-actin Distribution (2014)
  • Author(s): Nur Atik他
  • Journal Title: Cell Structure and Function Volume: 39 Issue: 1 Pages: 61-77
  • DOI: 10.1247/csf.13020
  • NAID: 130004137601
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed
• [Journal Article] CLAC-P/collagen type XXV is required for the intramuscular innervation of motoneurons during neuromuscular development. (2014)
  • Author(s): Tanaka T, Wakabayashi T, Oizumi H, Nishio S, Sato T, Harada A, Fujii D, Matsuo Y, Hashimoto T, Iwatsubo T.
  • Journal Title: J Neurosci. Volume: 34 Issue: 4 Pages: 1370-1379
  • DOI: 10.1523/jneurosci.2440-13.2014
  • Peer Reviewed
• [Journal Article] Hypothalamic SIRT1 prevents age-associated weight gain by improving leptin sensitivity in mice. (2014)
  • Author(s): Sasaki T, Kikuchi O, Shimpuku M, Susanti VY, Yokota-Hashimoto H, Taguchi R, Shibusawa N, Sato T, Tang L, Amano K, Kitazumi T, Kuroko M, Fujita Y, Maruyama J, Lee YS, Kobayashi M, Nakagawa T, Minokoshi Y, Harada A, Yamada M, Kitamura T.
  • Journal Title: Diabetologia Volume: 57 Issue: 4 Pages: 819-831
  • DOI: 10.1007/s00125-013-3140-5
  • Peer Reviewed / Open Access
• [Journal Article] Drebrin E depletion in human intestinal epithelial cells mimics Rab8a loss of function. (2014)
  • Author(s): Vacca B.他
  • Journal Title: Hum Mol Genet Volume: 23 Issue: 11 Pages: 2834-46
  • DOI: 10.1093/hmg/ddt670
  • Peer Reviewed
• [Journal Article] Trans-regulation of oligodendrocyte myelination by neurons through small GTPase Arf6-regulated secretion of fibroblast growth factor-2. (2014)
  • Author(s): Akiyama M.他
  • Journal Title: Nat Commun. Volume: 5 Issue: 1 Pages: 4744-4744
  • DOI: 10.1038/ncomms5744
  • Peer Reviewed
• [Journal Article] Caenorhabditis elegans chaperonin CCT/TRiC is required for actin and tubulin biogenesis and microvillus formation in intestinal epithelial cells (2014)
  • Author(s): Keiko Saegusa, Miyuki Sato, Katsuya Sato, Junko Nakajima-Shimada, Akihiro Harada, and Ken Sato
  • Journal Title: Mol. Biol. Cell Volume: 25 Issue: 20 Pages: 3095-3104
  • DOI: 10.1091/mbc.e13-09-0530
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Rab11a is required for apical protein localisation in the intestine. (2014)
  • Author(s): Sobajima T, Yoshimura S, Iwano T, Kunii M, Watanabe M, Atik N, Mushiake S, Morii E, Koyama Y, Miyoshi E, Harada A
  • Journal Title: Biol Open Volume: 4 Issue: 1 Pages: 86-94
  • DOI: 10.1242/bio.20148532
  • Peer Reviewed / Open Access
• [Journal Article] Rab8a and Rab8b are essential for multiple apical transport pathways but insufficient for ciliogenesis. (2013)
  • Author(s): Sato T, Iwano T, Kunii M, Matsuda S, Mizoguchi R, Jung Y, Hagiwara H, Yoshihara Y, Yuzaki M, Harada R, Harada A.
  • Journal Title: J Cell Sci. Volume: 15;127(Pt 2) Pages: 422-31
  • DOI: 10.1242/jcs.136903
  • Peer Reviewed / Open Access
• [Journal Article] Vesicular and non-vesicular transport feed distinct glycosylation pathways in the Golgi. (2013)
  • Author(s): D'Angelo G, Uemura T, Chuang CC, Polishchuk E, Santoro M, Ohvo-Rekila H, Sato T, Di Tullio G, Varriale A, D'Auria S, Daniele T, Capuani F, Johannes L, Mattjus P, Monti M, Pucci P, Williams RL, Burke JE, Platt FM, Harada A, De Matteis MA.
  • Journal Title: Nature Volume: 501 Issue: 7465 Pages: 116-120
  • DOI: 10.1038/nature12423
  • Peer Reviewed
• [Journal Article] Critical roles of type III phosphatidylinositol phosphate kinase in murine embryonic visceral endoderm and adult intestine (2012)
  • Author(s): Takasuga , Wada et al.
  • Journal Title: PNAS 110 Volume: 1726-1731 Issue: 5 Pages: 1726-31
  • DOI: 10.1073/pnas.1213212110
  • Peer Reviewed
• [Journal Article] Uncovering genes required for neuronal morphology by morphology-based gene trap screening with a revertible retrovirus vector. (2012)
  • Author(s): Hashimoto Y
  • Journal Title: FASEB Journal Volume: 26 Issue: 11 Pages: 4662-4674
  • DOI: 10.1096/fj.12-207530
  • Peer Reviewed
• [Journal Article] Delivery of endosomes to lysosomes via microautophagy in the visceral endoderm of mouse embryos (2012)
  • Author(s): Kawamura, Wada et al.
  • Journal Title: Nature Communications Volume: 3 Issue: 1 Pages: 1071-1071
  • DOI: 10.1038/ncomms2069
  • Peer Reviewed
• [Presentation] 細胞の極性形成における細胞内小胞輸送の役割 (2015)
  • Author(s): 吉村信一郎
  • Organizer: 第120回日本解剖学会総会・全国学術集会、第92回日本生理学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-03-22
• [Presentation] Analyses of neuronal and epithelial cell polarity of mice lacking genes involved in polarized transport. (2014)
  • Author(s): 原田彰宏
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-13
• [Presentation] 膵外分泌・内分泌における膜融合関連分子SNAP23の機能の解明 (2014)
  • Author(s): 國井政孝、吉村信一郎、高橋倫子、小林雅樹、佐藤隆史、川上良介、根本知己、河西晴郎、北村忠弘、佐藤健、原田彰宏
  • Organizer: 第66回日本細胞生物学大会
  • Place of Presentation: 奈良
  • Year and Date: 2014-06-13
• [Presentation] 表現型を正常に戻すことが可能なレトロウィルスベクターを用いた、形態によるスクリーニングによる神経細胞の形態形成に必要な遺伝子の同定 (2013)
  • Author(s): 原田彰宏
  • Organizer: 第６５回日本細胞生物学会大会
  • Place of Presentation: 名古屋
• [Presentation] 組織において、細胞内極性を司る分子は細胞の極性や分泌などにどのように関わるか？ (2013)
  • Author(s): 原田彰宏
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Invited
• [Presentation] Roles of membrane trafficking in cell polarity and secretion in model animals (2012)
  • Author(s): 原田彰宏
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡(招待講演)
• [Remarks] http://www.med.osaka-u.ac.jp/pub/acb/