聴・平衡覚器官のイメージング解析～１分子から個体レベルまで～

• Project Area: Mutli-dimensional fluorescence live imaging of cellular function and molecular activity
• Project/Area Number: 25113715
• 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: Kobe University
• Principal Investigator: 齋藤 尚亮 神戸大学, バイオシグナル研究センター, 教授 (60178499)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥10,660,000 (Direct Cost: ¥8,200,000、Indirect Cost: ¥2,460,000); Fiscal Year 2014: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
• Keywords: Cdc42 / 内耳 / 有毛細胞 / イメージング / 難聴 / アクチン

Outline of Annual Research Achievements

内耳有毛細胞特異的Cdc42 KOマウスが、1)「正常聴覚獲得後に多彩な聴毛の形態異常による緩徐進行性の75dB程度の高度難聴を呈する」、2)「Cdc42は聴毛の形成過程よりむしろ維持期に機能する」、3)「Cdc42は聴毛膜に局在する」 、4)「その活性化は聴毛の上半（先端）部の方が下半（基）部よりも強い（FRET法により証明）」等の４つの事実を世界に先駆けて明らかにした。
聴毛の形態異常については、欠損・短縮・伸長・癒合など所見を電子顕微鏡（図中矢印）にて明らかにした。このような聴毛の形態変化を引き起こす原因として、正常では聴毛及び頂側結合複合体（apical junctional complexes）に局在・活性化して、アクチン代謝を巧妙に調節しているCdc42が機能しないため、N-WASPによるアクチン重合とcofilinによるアクチン脱重合の両者が障害され、アクチン代謝が非常に低い状態で保たれるようになるためであることを証明した。

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] The Extracellular A-loop of Dual Oxidases Affects the Specificity of Reactive Oxygen Species Release. (2015)
  • Author(s): Ueyama T, Sakuma M, Ninoyu Y, Hamada T, Dupuy C, Geiszt M, Leto TL, Saito N.
  • Journal Title: J Biol. Chem Volume: 290 Issue: 10 Pages: 6495-6506
  • DOI: 10.1074/jbc.m114.592717
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification and characterization of PKCγ, a kinase associated with SCA14, as an amyloidogenic protein. (2015)
  • Author(s): Takahashi H, Adachi N, Shirafuji T, Danno S, Ueyama T, Vendruscolo M, Shuvaev AN, Sugimoto T, Seki T, Hamada D, Irie K, Hirai H, Sakai N, *Saito N.
  • Journal Title: Hum Mol Genet. Volume: 24 Issue: 2 Pages: 525-539
  • DOI: 10.1093/hmg/ddu472
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The role of Pak-interacting exchange factor-β phosphorylation at serines 340 and 583 by PKCγ in dopamine release. (2014)
  • Author(s): Shirafuji T, Ueyama T, Yoshino K, Takahashi H, Adachi N, Ago Y, Koda K, Nashida T, Hiramatsu N, Matsuda T, Toda T, Sakai N, Saito N.
  • Journal Title: J Neurosci Volume: 34 Issue: 28 Pages: 9268-9280
  • DOI: 10.1523/jneurosci.4278-13.2014
  • NAID: 120005619641
  • Peer Reviewed / Open Access
• [Journal Article] Maintenance of stereocilia and apical junctional complexes by Cdc42 in cochlear hair cells (2014)
  • Author(s): Ueyama T, Sakaguchi H, Nakamura T, Goto A, Morioka S, Shimizu A, Nakao K, Hishikawa Y, Ninoyu Y, Kassai H, Suetsugu S, Koji T, Fritzsch B, Yonemura S, Hisa Y, Matsuda M, Aiba A, Saito N.
  • Journal Title: J Cell Sci. Volume: 127 Pages: 2040-2052
  • DOI: 10.1242/jcs.143602
  • NAID: 120006986582
  • Peer Reviewed / Open Access
• [Journal Article] Heparin inhibits melanosome uptake and inflammatory response coupled with phagocytosis through blocking PI3k/Akt and MEK/ERK signaling pathways in human epidermal keratinocytes. (2014)
  • Author(s): Makino-Okamura C, Niki Y, Takeuchi S, Nishigori C, Declercq L, Yaroch DB, Saito N.
  • Journal Title: Pigment Cell Melanoma Res. Volume: 27 Issue: 6 Pages: 1063-1074
  • DOI: 10.1111/pcmr.12287
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] RANTES/CCL5 mediated-biological effects depend on the syndecan-4/PKCa signaling pathway (2014)
  • Author(s): Maillard, L., Saito, N., Hlawaty, H., Friand, V., Suffee, N., Chmilewsky, F., Haddad, O., Laguillier, C., Guyot, E., Ueyama, T., Oudar, O., Sutton, A., Charnaux, N.
  • Journal Title: Biology Open Volume: 3 Issue: 10 Pages: 995-1004
  • DOI: 10.1242/bio.20148227
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Both the C1 domain and a basic amino acid cluster at C-terminus are important for neurite and branch induction ability of DGK. (2014)
  • Author(s): Takuya Kano, Takeshi Kouzuki, Satoru Mizuno, Shuji Ueda, Minoru Yamanoue, Fumio Sakane, Naoaki Saito and Yasuhito Shirai
  • Journal Title: iochem. Biophys. Res. Commun Volume: 印刷中 Issue: 1 Pages: 89-94
  • DOI: 10.1016/j.bbrc.2014.03.113
  • Peer Reviewed / Open Access
• [Journal Article] Diacylglycerol kinase gamma regulates antigen-induced mast cell degranulation by mediating Ca2+ influxes (2014)
  • Author(s): Sakuma, M., Shirai, Y., Ueyama, T., and Saito, N.
  • Journal Title: Biochem. Biophys. Res. Commun, Volume: 445 Issue: 2 Pages: 340-345
  • DOI: 10.1016/j.bbrc.2014.01.197
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Deregulation of the actin cytoskeleton and macropinocytosis in response to phorbol ester by the mutant protein kinase C gamma that causes spinocerebellar ataxia type 14. (2014)
  • Author(s): Yamamoto K, Seki T, Yamamoto H, Adachi N, Tanaka S, Hide I, Saito N, Sakai N
  • Journal Title: Front Physiol. Volume: 5 Pages: 126-126
  • DOI: 10.3389/fphys.2014.00126
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Diacylglycerol kinase as a possible therapeutic target for neuronal diseases. (2014)
  • Author(s): Shirai, Y and Saito, N
  • Journal Title: Journal of Biomedical Science Volume: 21 Pages: 28-36
  • Peer Reviewed
• [Journal Article] Maintenance of stereocilia and apical junctional complexes by Cdc42 in cochlear hair cells, (2014)
  • Author(s): Ueyama, T., Sakaguchi, H., Nakamura, T., Goto, A., Morioka, S., Shimizu, A., Nakao, K., Hishikawa, Y., Ninoyu, Y., Kassai, H., Suetsug, J., Koji, T., Fritzsch, B., Yonemura, S., Hisa, Y., Matsuda, M., Aiba, A., and Saito, N.
  • Journal Title: J. Cell Science Volume: 印刷中
  • NAID: 120006986582
  • Peer Reviewed
• [Journal Article] Negative Charges in the Flexible N-Terminal Domain of Rho GDP-Dissociation Inhibitors (RhoGDIs) Regulate the Targeting of the RhoGDI-Rac1 Complex to Membranes (2013)
  • Author(s): Ueyama T, Son J, Kobayashi T, Hamada T, Nakamura T, Sakaguchi H, Shirafuji T, Saito N
  • Journal Title: J. Immunol. Volume: 191 Issue: 5 Pages: 2560-2569
  • DOI: 10.4049/jimmunol.1300209
  • Peer Reviewed
• [Journal Article] Mutant gammaPKC that causes spinocerebellar ataxia type 14 upregulates Hsp70, which protects cells from the mutant's cytotoxicity. (2013)
  • Author(s): Ogawa, K., Seki, T., Onji, T., Adachi, N., Tanaka, S., Hide, I., Saito, N., Sakai, N.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 440 Issue: 1 Pages: 25-30
  • DOI: 10.1016/j.bbrc.2013.09.013
  • Peer Reviewed
• [Journal Article] PKC-ε pseudosubstrate and catalytic activity are necessary for membrane delivery during IgG-mediated phagocytosis. (2013)
  • Author(s): Wood, TR., Chow, RY., Hanes, CM., Zhang, X., Kashiwagi, K., Shirai, Y., Trebak, M., Loegering, DJ., Saito, N., and Lennartz, MR.
  • Journal Title: J Leukoc Biol Volume: 94 Issue: 1 Pages: 109-122
  • DOI: 10.1189/jlb.1212634
  • Peer Reviewed
• [Journal Article] Increased seizure susceptibility in a mouse with diacylglycerol kinase β 2 deficiency. (2013)
  • Author(s): Ishisaka, M., Tsuruma, K., Shimazawa, M., Shirai, Y., Saito, N., and Hara, H.
  • Journal Title: Neuroscience & Medicine. Volume: 4 Pages: 117-122
  • Peer Reviewed
• [Journal Article] Protein kinase Cη is targeted to lipid droplets. (2013)
  • Author(s): Suzuki Michitaka
  • Journal Title: Histochem Cell Biol Volume: 139 Issue: 4 Pages: 505-511
  • DOI: 10.1007/s00418-013-1083-z
  • Peer Reviewed
• [Journal Article] Both C1B domain and pseudosubstrate region are necessary for saturated fatty acid membrane;distinct role of intramolecular domains for different (2013)
  • Author(s): Takaaki Nishimoto, T., Kashiwagi, K., Saito, N., and Shirai, Y.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 432 Issue: 2 Pages: 384-388
  • DOI: 10.1016/j.bbrc.2013.01.048
  • Peer Reviewed
• [Presentation] 内耳有毛細胞におけるRhoファミリー低分子量G蛋白質の役割 (2014)
  • Author(s): 上山健彦、齋藤尚亮
  • Organizer: 日本組織細胞化学会
  • Place of Presentation: 松本
  • Year and Date: 2014-09-27 – 2014-09-28
• [Presentation] Cdc42 is required for maintenance of stereocilia and apical junctional complexes in cochlear hair cells. (2014)
  • Author(s): Ueyama, T., Sakaguchi, H., Goto, A., Hisa, Y., Matsuda, M., Aiba, A. and Saito, N.
  • Organizer: The 37th Naito Conference
  • Place of Presentation: 北海道ニセコ
  • Year and Date: 2014-07-15 – 2014-07-18
• [Presentation] 内耳有毛細胞特異的Cdc42ノックアウトマウスは聴毛及び頂側細胞間接着時の アクチン代謝障害により緩徐進行性の高度難聴を呈する (2014)
  • Author(s): 中村 高志、上山 健彦、坂口 博史、後藤 明弘、森岡 繁文、米村 重信、 久 郁男、松田 道行、饗庭 篤、齋藤 尚亮
  • Organizer: 第87回日本薬理学会年会
  • Place of Presentation: 仙台国際センター、仙台
• [Presentation] Protein kinase C and its involvement in neurodegenerative disease. (2014)
  • Author(s): Naoaki Saito
  • Organizer: Symposium on Membrane Biology
  • Place of Presentation: Seattle, WA, USA
  • Invited
• [Presentation] A protein kinase Cgamma knockout mice as a Parkinsonian model: Involvement of Pak -Interacting Exchange Factorbeta. (2014)
  • Author(s): 齋藤尚亮
  • Organizer: 第87回日本薬理学会年会
  • Place of Presentation: 仙台国際センター、仙台
• [Presentation] Cde-42 is required for maintenance of stereocilia and apical junctional complexes in cochlear hair cells. (2014)
  • Author(s): Ueyama, T. and Saito, N
  • Organizer: Joint Workshop between University Catholic Louvain and Kobe University
  • Place of Presentation: Brussels, Belgium
• [Presentation] Both C1 domain and basic amino acid cluster at C-terminus are important for branching and neurite induction of DGKbeta (2013)
  • Author(s): Shirai,Y., Takuya,K., Kouzuki,T., Ueda,S., Yamanoue,M., Sakane,F. and Saito,N
  • Organizer: Neuro 2013
  • Place of Presentation: 京都国際会館、京都