Production and analysis of model animals for neurodegenerative diseases caused by dysfunction of PKC and their use for drug design

• Project/Area Number: 21390070
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General pharmacology
• Research Institution: Kobe University
• Principal Investigator: SAITO Naoaki 神戸大学, バイオシグナル研究センター, 教授 (60178499)
• Co-Investigator(Kenkyū-buntansha): SHIRAI Yasuhito 神戸大学, バイオシグナル研究センター, 准教授 (60263399) ; UEYAMA Takehiko 神戸大学, バイオシグナル研究センター, 准教授 (80346254)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥18,590,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥4,290,000); Fiscal Year 2011: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2010: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2009: ¥8,450,000 (Direct Cost: ¥6,500,000、Indirect Cost: ¥1,950,000)
• Keywords: プロテインキナーゼ / 神経変性疾患 / モデル動物 / 創薬 / 細胞内情報伝達 / プロテインキナーゼC / 脊髄小脳変性症 / 神経変性 / カルシウム / 遺伝子変異 / アミロイド

Research Abstract

Functional involvement of PKCγin the etiology of SCA14 and Parkinson disease was studied in cells and animals. We found 1) mutant PKCγs form amyloid-like fibrils in Purkinje cells and the formation was inhibitied by trehalose, 2) dysfunction of PKCγcould cause Parkinson disease by the inhibition of dopamine release and the loss of dopaminergic neurons in the substantia nigra.

Research Products

• [Journal Article] MARCKS dephosphorylation is involved in bradykinin-induced neurite outgrowth in neuroblastoma SH-SY5Y cells (2012)
  • Author(s): Tanabe A, Shiraishi M, Negishi M, Saito N, Tanabe M, and Sasaki Y.
  • Journal Title: J Cell Physiol Volume: 227(2) Pages: 618-629
• [Journal Article] Establishment of a Novel Fluorescence-Based Method to Evaluate Chaperone-Mediated Autophagy in a Single Neuron (2012)
  • Author(s): Seki, T., Ken-ich Yoshino, K., Tanaka, S., Dohi, E., Onji, T., Yamamoto, K., Hide, I., Paulson, H. L., Saito, N., Sakai, N.
  • Journal Title: PLoS ONE Volume: 7(2)
• [Journal Article] c-Abl tyrosine kinase regulates serum-induced nuclear export of diacylglycerol kinaseαby phosphorylation at Tyr218 (2012)
  • Author(s): Matsubara, T., Ikeda, M., Kiso, Y., Sakuma, M., Yoshino, K-I., Sakane, F., Merida, I., Saito, N. and Shirai, Y.
  • Journal Title: J. Biol. Chem Volume: 287 Pages: 5507-5517
• [Journal Article] MARCKS dephosphorylation is involved in bradykinin-induced neurite outgrowth in neuroblastoma SH-SY5Y cells (2012)
  • Author(s): Tanabe A, Shiraishi M, Negishi M, Saito N, Tanabe M, Sasaki Y
  • Journal Title: J Cell Physiol Volume: 227 Pages: 618-629
  • Peer Reviewed
• [Journal Article] Establishment of a Novel Fluorescence-Based Method to Evaluate Chaperone-Mediated Autophagy in a Single Neuron (2012)
  • Author(s): Seki, T., Yoshino, K., Tanaka, S., Dohi, E., Onji, T., Yamamoto, K., Hide, I., Paulson, H.L., Saito, N., Sakai, N.
  • Journal Title: PLoS ONE Volume: 7
  • Peer Reviewed
• [Journal Article] c-Abl tyrosine kinase regulates serum-induced nuclear export of diacylglycerol kinase α by phosphorylation at Tyr218 (2012)
  • Author(s): Matsubara, T., Ikeda, M., Kiso, Y., Sakuma, M., Yoshino, K-I., Sakane, F., Merida, I., Saito, N., Shirai, Y.
  • Journal Title: J.Biol.Chem. Volume: 287 Pages: 5507-5517
  • Peer Reviewed
• [Journal Article] Cooperation of p40phox with p47phox for Nox2 activation during Fc{lower case gamma} R-mediated phagocytosis-Mechanism for acquisition of p40phox PI(3) P binding- (2011)
  • Author(s): Ueyama, T., Nakakita, J., Kobayashi, T., Kobayashi, T., Son, J-h., Sakaguchi, H., Leto, T., and Saito, N.
  • Journal Title: J. Biol. Chem Volume: 286 Pages: 40693-40705
• [Journal Article] Molecular pathophysiology of neurodegenerative disease caused byγPKC mutations (2011)
  • Author(s): Sakai, N., Saito, N. and Seki, T.
  • Journal Title: World J. Biological Psychiatry Volume: 12(S1) Pages: 95-98
• [Journal Article] Elucidation of the Molecular Mechanism and Exploration of Novel Therapeutics for Spinocerebellar Ataxia Caused by Mutant Protein Kinase Cγ (2011)
  • Author(s): Seki, T., Adachi, N., Abe-Seki, N., Shimahara, T., Takahashi, H., Yamamoto, K., Saito, N. and Sakai, N.
  • Journal Title: J Pharmacol Sci Volume: 116 Pages: 239-247
  • NAID: 10029894921
• [Journal Article] Streptococcus mutans diacylglycerol kinase homologue : a potential target for anti-caries chemotherapy (2011)
  • Author(s): Shibata, Y., Kawada-Matsuo, M., Shirai, Y., Saito, N., Li. D., and Yamashita, Y.
  • Journal Title: J. Med. Microbiol Volume: 60 Pages: 625-630
  • NAID: 120004951599
• [Journal Article] Direct binding of RalA to PKCh and its crucial role in morphological change during keratinocytes differentiation (2011)
  • Author(s): Shirai, Y., Morioka, S., Sakuma, M., Yoshino, K., Otsuji, C., Sakai, N., Kashiwagi, K., Chida, K., Shirakawa, R., Horiuchi, H., Nishigori, C., Ueyama, T., Saito, N.
  • Journal Title: Mol. Biol. Cell Volume: 22 Pages: 1340-1352
• [Journal Article] Nuclear Choline Acetyltransferase Activates Transcription of a High-affinity Choline Transporter (2011)
  • Author(s): Matsuo, A., Bellier, J-P, Nishimura, M., Yasuhara, O., Saito, N. and Kimura, H.
  • Journal Title: J. Biol. Chem Volume: 286 Pages: 5836-5845
• [Journal Article] Cooperation of p40phox with p47phox for Nox2 activation during Fc{lower case gamma} R-mediated phagocytosis--Mechanism for acquisition of p40phox PI(3)P binding-- (2011)
  • Author(s): Ueyama, T., Nakakita, J., Kobayashi, T., Kobayashi, T.,-hyun Son, J-h., Sakaguchi, H., Leto, T., Saito, N.
  • Journal Title: J.Biol.Chem. Volume: 286 Pages: 40693-40705
  • Peer Reviewed
• [Journal Article] Molecular pathophysiology of neurodegenerative disease caused by γPKC mutations (2011)
  • Author(s): Sakai, N., Saito, N., Seki, T.
  • Journal Title: World J.Biological Psychiatry Volume: 12 Pages: 95-98
  • Peer Reviewed
• [Journal Article] Streptococcus mutans diacylglycerol kinase homologue : a potential target for anti-caries chemotherapy (2011)
  • Author(s): Shibata, Y., Kawada-Matsuo, M., Shirai, Y, Saito, N., Li.D., Yamashita, Y.
  • Journal Title: J.Med.Microbiol. Volume: 60 Pages: 625-630
  • NAID: 120004951599
  • Peer Reviewed
• [Journal Article] Direct binding of RalA to PKCη and its crucial role in morphological change during keratinocytes differentiation (2011)
  • Author(s): Shirai, Y., Morioka, S., Sakuma, M., Yoshino, K., Otsuji, C., Sakai, N., Kashiwagi, K., Chida, K., Shirakawa, R., Horiuchi, H., Nishigori, C., Ueyama, T., Saito, N.
  • Journal Title: Mol.Biol.Cell Volume: 22 Pages: 1340-1352
  • Peer Reviewed
• [Journal Article] Nuclear Choline Acetyltransferase Activates Transcription of a High-affinity Choline Transporter (2011)
  • Author(s): Matsuo, A., Bellier, J-P, Nishimura, M., Yasuhara, O., Saito, N., Kimura, H.
  • Journal Title: J.Biol.Chem. Volume: 286 Pages: 5836-5845
  • Peer Reviewed
• [Journal Article] Diacylglycerol kinaseβknockout mice exhibit lithium-sensitive behavioral abnormalities (2010)
  • Author(s): Kakefuda K., Oyagi A., Ishisaka M., Tsuruma K., Shimazawa M., Yokota K., Shirai Y., Horie K., Saito N., Takeda J. and Hara H.
  • Journal Title: PLoS One Volume: 5(10)
• [Journal Article] A critical role of conventional protein kinase C in morphological changes of rodent mast cells (2010)
  • Author(s): Yanase Y., Hide, I., Mihara, S., Shirai, Y., Saito, N., Nakata, Y., Hide, M., and Sakai, N.
  • Journal Title: Immunol. Cell. Biol Volume: 89(1) Pages: 149-59
• [Journal Article] Supression mechanisms of flavonoids on aryl hydrocarbon receptor-mediated signal transduction (2010)
  • Author(s): Mukai, R., Shirai, Y., Saito, N., Fukuda, I., Nishiumi, S., Yoshida, K., Ashida, H.
  • Journal Title: Arch. Biochem. Biophys Volume: 501 Pages: 134-141
• [Journal Article] The effect of trehalose on the properties of mutantγPKC, which causes spinocerebellar ataxia type 14(SCA14), in neuronal cell lines and cultured Purkinje cells (2010)
  • Author(s): Seki, T., Abe-Seki, N., Kikawada, T., Takahashi, H., Yamamoto, K., Adachi, N., Tanaka, S., Hide, I., Saito, N., and Sakai, N.
  • Journal Title: J. Biol. Chem Volume: 285 Pages: 33252-33264
• [Journal Article] PKC-induced phosphorylation of Orai1 regulates intracellular Ca2+ level via store operated Ca2+ channel (2010)
  • Author(s): Kawasaki, T., Ueyama, T., Lange, I., Feske, S. Saito, N.
  • Journal Title: J. Biol. Chem Volume: 285(33) Pages: 25720-30
• [Journal Article] Essential role of diacylglycerol kinase beta(DGKb) in neurite spine formation, contributing to cognitive function including spatial and long-term memory (2010)
  • Author(s): Shirai, Y., Kozuki, T., Kakefuda, K., Moriguchi, S., Oyagi, A., Horie, K., Shimazawa, M., Fukunaga, K., Takeda, J., Saito, N., Hara, H.
  • Journal Title: PLoS One Volume: 5(7)
• [Journal Article] Mutant protein kinase C gamma that causes spinocerebellar ataxia type 14(SCA14) is selectively degraded by autophagy (2010)
  • Author(s): Yamamoto, K., Seki, T., Adachi, N., Takahashi, T., Tanaka, S., Hide, I., Saito, N., Sakai, N.
  • Journal Title: Genes to Cells Volume: 15 Pages: 425-38
• [Journal Article] Potential Antitumor Activity of a Low Molecular Weight Protein Fraction(MLP-Fraction) from Grifola frondosa through Enhancement of Cytokine Production (2010)
  • Author(s): Kodama, N., Mizuno, S., Nanba, H. and Saito, N.
  • Journal Title: Journal of Medicinal Food Volume: 13 Pages: 20-30
• [Journal Article] Stimulatory effects of the soy phytoestrogen genistein on noradrenalin transporter and serotonin transporter activity (2010)
  • Author(s): Toyohira, Y., Ueno, S., Tsutsui, M., Itho, H., Sakai, N., Saito, N., Takahashi, K., and Yanagihara, N.
  • Journal Title: Mol. Nutr. Food Res Volume: 54 Pages: 516-524
• [Journal Article] Diacylglycerol kinase β knockout mice exhibit lithium-sensitive behavioral abnormalities. (2010)
  • Author(s): Kakefuda K, et al.
  • Journal Title: PLoS One Volume: 5(10)
  • Peer Reviewed
• [Journal Article] Supression mechanisms of flavonoids on aryl hydrocarbon receptor-mediated signal transduction. (2010)
  • Author(s): Mukai, R., et al.
  • Journal Title: Arch.Biochem.Biophys Volume: 501 Pages: 134-141
  • Peer Reviewed
• [Journal Article] The effect of trehalose on the properties of mutant γPKC, which causes spinocerebellar ataxia type 14 (SCA14), in neuronal celllines and cultured Purkinie cells. (2010)
  • Author(s): Seki, T., et al.
  • Journal Title: J.Biol.Chem Volume: 285 Pages: 33252-264
  • Peer Reviewed
• [Journal Article] PKC-induced phosphorylation of Orai1 regulates intracellular Ca2+level via store operated Ca2+channel. (2010)
  • Author(s): Kawasaki, T., et al
  • Journal Title: J.Biol.Chem Volume: 285 Pages: 25720-30
  • Peer Reviewed
• [Journal Article] Essential role of diacylglycerol kinase beta (DGKb) in neurite spine formation, contributing to cognitive function including spatial and long-term memory. (2010)
  • Author(s): Shirai, Y., et al.
  • Journal Title: PLoS One Volume: 5(7)
  • Peer Reviewed
• [Journal Article] Potential Antitumor Activity of a Low Molecular Weight Protein Fraction(MLP-Fraction)from Grifola frondosa through Enhancement of Cytokine Production (2010)
  • Author(s): Kodama, N., et al
  • Journal Title: Journal of Medicinal Food 13 Pages: 20-30
  • Peer Reviewed
• [Journal Article] Inflammatory Cytokine TNF-alpha enhances NGF Production in Human Keratinocytes, HaCaT Cells (2010)
  • Author(s): Takaoka, K., et al
  • Journal Title: J.Pharmacol.Sci. 111 Pages: 381-391
  • Peer Reviewed
• [Journal Article] Mutant protein kinase C gamma that causes spinocerebellar ataxia type 14(SCA14)is selectively degraded by autophagy (2010)
  • Author(s): Yamamoto, K., et al
  • Journal Title: Genes to Cells (印刷中)
  • Peer Reviewed
• [Journal Article] Stimulatory effects of the soy phytoestrogen genistein on noradrenalin transporter and serotonin transporter activity (2010)
  • Author(s): Toyohira, Y., et al
  • Journal Title: Mol.Nutr.Food Res. (印刷中(掲載確定))
  • Peer Reviewed
• [Journal Article] Duox maturation factors form cell surface complexes with Duox affecting the specificity of reactive oxygen species generation (2009)
  • Author(s): Morand S, et al
  • Journal Title: FASEB J. 23 Pages: 1205-1218
  • Peer Reviewed
• [Journal Article] Mutant γPKC found in spinocerebellar ataxia type 14 induces aggregate-independent maldevelopment of dendrites in primary cultured Purkinje cells (2009)
  • Author(s): Seki, T, et al
  • Journal Title: Neurobiology of Disease 33 Pages: 260-273
  • Peer Reviewed
• [Journal Article] A Simple Analog of Tumor-promoting Aplysiatoxin Is an Antineo plastic Agent rather than a Tumor Promoter : Development of a Synthetically Accessible PKC Activator with Bryostatin-like Activity (2009)
  • Author(s): Nakagawa, Y., et al.
  • Journal Title: J Am.Chem.Soc. 131 Pages: 7573-7579
  • Peer Reviewed
• [Journal Article] Kinase activity of the dgk gene product is involved 1 in the virulence of Streptococcus mutans (2009)
  • Author(s): Shibata, Y., et al
  • Journal Title: Microbiology 155 Pages: 557-565
  • Peer Reviewed
• [Journal Article] A technique for monitoring multiple signals with a combination of prism-based total internal reflection fluorescence microscopy and epifluorescence microscopy (2009)
  • Author(s): Adachi, E., et al
  • Journal Title: Pflugers Arch 459 Pages: 227-234
  • Peer Reviewed
• [Journal Article] Subcellular localization of flavonol aglycone inhepatocytes visualized by confocal laser scanning fluorescent microscope (2009)
  • Author(s): Mukai, R., et al
  • Journal Title: Cytotechnology 59 Pages: 177-182
  • Peer Reviewed
• [Journal Article] Direct binding of RaIA to PKCη and its crucial role in morphological change during keratinocytes differentiation.
  • Author(s): Shirai, Y., et al.
  • Journal Title: Mol.Biol.Cell Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] Nuclear Choline Acetyltransferase Activates Transcription of a High-affinity Choline Transporter.
  • Author(s): Matsuo, A., et al.
  • Journal Title: J.Biol.Chem Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] A critical role of conventional protein kinase C in morphological changes of rodent mast cells.
  • Author(s): Yanase Y., et al
  • Journal Title: Immunol.Cell.Biol. Volume: (印刷中)
  • Peer Reviewed
• [Presentation] 細胞内シグナルの可視化と細胞機能 (2012)
  • Author(s): 齋藤尚亮
  • Organizer: 第89日本生理学会大会
  • Place of Presentation: (長野県)松本文化会館
  • Year and Date: 2012-03-31
• [Presentation] 中枢神経損傷とRho family GTPases (2012)
  • Author(s): 上山健彦、齋藤尚亮
  • Organizer: 第85回日本薬理学会年会
  • Place of Presentation: (京都府)国立京都国際会館
  • Year and Date: 2012-03-15
• [Presentation] 創薬ターゲットとしてのジアシルグリセロールキナーゼ (2012)
  • Author(s): 白井康仁、齋藤尚亮
  • Organizer: 第85回日本薬理学会年会
  • Place of Presentation: (京都府)国立京都国際会館(口頭発表、日本語)
  • Year and Date: 2012-03-15
• [Presentation] 細胞内シグナル分子の可視化と細胞機能 (2012)
  • Author(s): 齋藤尚亮
  • Organizer: 第89回日本生理学大会
  • Place of Presentation: 松本
• [Presentation] Novel phosphorylation site(s) of cofilin by PKCα and its negative role in the degranulation from RBL-2H3 mast cells (2011)
  • Author(s): Sakuma, M., Shirai, Y., Yoshino K., Kuramasu, M., Nakamura, T., Mizuno, K., Hide, I., Nakata, Y., Saito, N.
  • Organizer: The American society for cell biology
  • Place of Presentation: デンバー、米国
• [Presentation] PKCgノックアウトは黒質線条体ドパミン神経系の異常をもたらす (2010)
  • Author(s): 白藤俊彦、足立直子、高橋英之、香田健、梨子田哲明、吾郷由希夫、松田敏夫、齋藤尚亮
  • Organizer: 第118回日本薬理学会近畿部会
  • Place of Presentation: 大阪
  • Year and Date: 2010-11-19
• [Presentation] Visualization of PKC targeting and its involvement in the pathogenesis of spinocerebellar ataxia (2010)
  • Author(s): Saito, N.
  • Organizer: 第51回日本組織細胞化学会
  • Place of Presentation: 東京
• [Presentation] PKC targeting and its involvement in the pathogenesis of spinocerebellar ataxia (2010)
  • Author(s): Saito, N.
  • Organizer: 14^<th> International congress of Endocrinology
  • Place of Presentation: Kyoto, Japan
• [Presentation] Molecular pathophysiology of spinocerebellar ataxia type14(SCA14), a hereditary neurodegenerative disease caused byγPKC mutations (2009)
  • Author(s): Sakai, N., Seki, T., Adachi, N., Takahashi, H. and Saito, N.
  • Organizer: Japan-Taiwan joint symposium on Cell Sygnaling and Gene Regulation
  • Place of Presentation: Kobe, Japan
• [Remarks]
  • URL: http://www.research.kobe-u.ac.jp/brce-saito/