Imaging analysis reveals mechanistic differences between first and second phase insulin exocytosis

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 17590277
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pathological medical chemistry
• Research Institution: Kyorin University
• Principal Investigator: IMAIZUMI Mika Kyorin University, Faculty of Medicine, Associate professor (40201941)
• Co-Investigator(Kenkyū-buntansha): NAGAMATSU Shinya Kyorin University, Faculty of Medicine, Professor (80231489) ; AOYAGI Kyota Kyorin University, Faculty of Medicine, Instructor (50453527) ; KIKUTA Toshiteru Waseda University, Faculty of Science and Engineering, Assistant Professor (80267468)
• Project Period (FY): 2005 – 2007
• Keywords: insulin / exocytosis / pancreatic beta cell / TIRF microscopy / type 2 diaetes / secretory granule

Research Abstract

The mechanism of glucose-induced biphasic insulin release is unknown. We used total internal reflection fluorescence (TIRF) imaging analysis to reveal the process of first- and second-phase insulin exocytosis in pancreatic 13 cells. This analysis showed that previously docked insulin granules fused at the. site of syntaxin (Synt) 1A clusters during the first phase; however, the newcomers fused during the second phase external to the Synt1A clusters. In order to reveal the function of Synt1A in phasic insulin exocytosis, we generated Synt1A-knockout (Synt1A^<-/->) mice. Synt1A^<-/->β cells showed fewer previously docked granules with no fusion during the first phase; second-phase fusion from newcomers was well preserved. Rescue experiments using adenovirus-expressed Synt1A in Synt1A^<-/->β cells showed restoration of granule docking status and fusion events during the first phase. Inhibition of the functions of other plasma membrane syntaxins, Synt3 and Synt4, did not affect second-phase insulin exocytosis by experiments with dominant negative forms of Synt3 and Synt4. Thus, our findings show that the first phase is Synt1A-dependent, but the second phase is not. This indicates that insulin exocytosis underlying biphasic release differs spatially and mechanistically.

Research Products

• [Journal Article] Adiponectin induces insulin secretion in vitro and in vivo at a low glucose concentration (2008)
  • Author(s): Okamoto M., Ohara-Imaizumi M., Nagamatsu S.(他9名)
  • Journal Title: Diabetologia 51 Pages: 827-835
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Imaging of insulin exocytosis in GK rat beta cells subjected to long-term glinide or sulphonylurea treatment (2008)
  • Author(s): Kawai J., Ohara-Imaizumi M., Nagamatsu S.(他9名)
  • Journal Title: Biochem.J. 412 Pages: 93-101
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Adiponectin induces insulin secretion in vitro and in vivo at a low glucose concentration (2008)
  • Author(s): Okamoto, M*, Ohara-Imaizumi, M*, Kubota, N, Hashimoto, S, Eto, K, Kanno, T, Kubota, T, Wakui, M, Nagai, R, Noda, M, Nagamatsu, S, Kadowaki, T
  • Journal Title: Diabetologia 51 Pages: 827-835
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Imaging of insulin exocytosis in GK rat beta cells subjected to long-term glinide or sulphonylurea treatment (2008)
  • Author(s): Kawai, J., Ohara-Imaizumi, M., Nakamichi, Y., Okamura, T., Akimoto, Y., Matsushima, S., Aovagi, K., Kawakami, H., Watanabe, T., Watada, H., Kawamori, R., Nagamatsu, S
  • Journal Title: Biochem. J 412 Pages: 93-101
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Elevation of the post-translational modification of proteins by O-linked N-acetylglucosamine leads to deterioration of the glucose-stimulated insulin secretion in the pancreas of diabetic Goto-Kakizaki rats (2007)
  • Author(s): Akimoto, Y., Ohara-Imaizumi M., Nagamatsu S.(他8名)
  • Journal Title: Glycobiology 17 Pages: 127-140
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] EphA-Ephrin-A-mediated beta cell communication regulates insulin secretion from pancreatic islets (2007)
  • Author(s): Konstantinova, I., Ohara-Imaizumi M., Nagamatsu S.(他8名)
  • Journal Title: Cell 129 Pages: 359-370
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Imaging analysis reveals mechanistic differences between first- and second-phase insulin exocytosis (2007)
  • Author(s): Ohara-lmaizumi M., Nagamatsu S.(他9名)
  • Journal Title: J Cell Biol 177 Pages: 695-705
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Protein kinase Cdelta plays a non-redundant role in insulin secretion in pancreatic beta cells (2007)
  • Author(s): Uchida, T., Ohara-ImaizumiM., Nagamatsu S.(他8名)
  • Journal Title: J Biol Chem 282 Pages: 2707-2716
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] InsP_7 debut in insulin release (2007)
  • Author(s): Nagamatsu S., Ohara-Imaizumi M.
  • Journal Title: Science 318 Pages: 1249-1250
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Imaging exoocytosis of single insulin secretory granules with TIRF microscopy (2007)
  • Author(s): Nagamatsu S., Ohara-Imaizumi M.
  • Journal Title: Methods Mol.Biol 440 Pages: 259-268
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Elevation of the post-translational modification of proteins by O-linked N-acetylglucosamine leads to deterioration of the glucose-stimulated insulin secretion in the pancreas of diabetic Goto-Kakizaki rats (2007)
  • Author(s): Akimoto, Y., Hart, G W., Wells, L., Vosseller, K., Yamamoto, K., Munetomo, E., Ohara-Imaizumi, M., Nishiwaki, C., Nagamatsu, S., Hirano, H., Kawakami, H
  • Journal Title: Glycobiology 17 Pages: 127-140
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] EphA-Ephrin-A-mediated beta cell communication regulates insulin secretion from pancreatic islets (2007)
  • Author(s): Konstantinova, I., Nikolova, G, Ohara-Imaizumi, M., Meda, P., Kucera, T., Zarbalis, K., Wurst, W., Nagamatsu, S., Lammert, E
  • Journal Title: Cell 129 Pages: 359-370
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Imaging analysis reveals mechanistic differences between first- and second-phase insulin exocytosis (2007)
  • Author(s): Ohara-Imaizumi, M., Fujiwara, T., Nakamichi, Y., Okamura, T., Akimoto, Y., Kawai, J., Matsushima, S., Kawakami, H., Watanabe, T., Akagawa, K., Nagamatsu, S
  • Journal Title: J Cell Biol 177 Pages: 695-705
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Protein kinase Cdelta plays a non-redundant role in insulin secretion in pancreatic beta cells. J Biol Chem (2007)
  • Author(s): Uchida, T, Iwashita, N, Ohara-Imaizumi, M, Ogihara, T, Nagai, S, Choi, JB, Tamura, Y, Tada, N, Kawamori, R, Nakayama, KI, Nagamatsu, S, Watada, H
  • Journal Title: Jan 26 282 (4) Pages: 2707-2716
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] InsP7 debut in insulin release (2007)
  • Author(s): Nagamatsu, S., Ohara-Imaizumi, M
  • Journal Title: Science 318 Pages: 1249-1250
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Imaging exoocytosis of single insulin secretory granules with TIRF microscopy (2007)
  • Author(s): Nagamatsu, S., Ohara-Imaizumi, M
  • Journal Title: Methods Mol. Biol 440 Pages: 259-268
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Imaging docking and fusion of insulin granules induced by antidiabetes agents:sulfonylurea and glinide drugs preferentially mediate the fusion of newcomer,but not previously docked,insulin granules (2006)
  • Author(s): Nagamatsu S, , Ohara-Imaizumi M., (他3名)
  • Journal Title: Diabetes 55 Pages: 2819-2825
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Intracellular dynamics observed by mode switching of microscope with a light incidence to thevnterface at alternate angles through the ultra high NA objective (2006)
  • Author(s): Wakazono, Y., Ohara-Imaizumi M., Nagamatsu S.(他3名)
  • Journal Title: Proc.SPIE 6088 Pages: 449-454
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Imaging docking and fusion of insulin granules induced by antidiabetes agents : sulfonylurea and glinide drugs preferentially mediate the fusion of newcomer, but not previously docked, insulin granules (2006)
  • Author(s): Nagamatsu, S., Ohara-Imaizumi, M., Nakamichi, Y, Kikuta, T., Nishiwaki, C
  • Journal Title: Diabetes 55 Pages: 2819-2825
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Intracellular dynamics observed by mode switching of microscope with a light incidence to the interface at alternate angles through the ultra high NA objective (2006)
  • Author(s): Wakazono, Y., Sakurai, T., Ohara-Imaizumi, M., Nagamatsu, S., Yamamoto, S., Terakawa. S
  • Journal Title: Proc. SPIE 6088 Pages: 449-454
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] ELKS,a protein structurally related to the active zone-associated protein CAST,is expressed in pancreatic beta cells and functions in insulin exoc ytosis:interaction of ELKS with exocytotic machinery analyzed by total internal reflection fluorescence microscopy (2005)
  • Author(s): Ohara-Imaizumi M., Nagamatsu S.(他9名)
  • Journal Title: Mol Biol Cell 16 Pages: 3289-3300
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Rab27a mediates the fight docking of insulin granules onto the plasma membrane during glucose stimulation (2005)
  • Author(s): Kasai K, Ohara-ImaizumiM., Nagamatsu S.(他7名)
  • Journal Title: J Clin Invest 115 Pages: 388-396
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Docking and fusion of insulin secretory granules in SURI knock out mouse beta-cells observed by total internal reflection fluorescence microscopy (2005)
  • Author(s): Kikuta T, Ohara-ImaizumiM., NagamatsuS.(他7名)
  • Journal Title: FEBS Lett 579 Pages: 1602-1606
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Renin-angiotensin system modulates oxidative stress-induced endothelial cell anontosis in rats (2005)
  • Author(s): Akishita M., Ohara-Imaizumi M., Nagamatsu S.(他8名)
  • Journal Title: Hypertension 45 Pages: 1188-1193
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] ELKS, a protein structurally related to the active zone-associated protein CAST, is expressed in pancreatic beta cells and functions in insulin exocytosis : interaction of ELKS with exocytotic machinery analyzed by total internal reflection fluorescence microscopy (2005)
  • Author(s): Ohara-Imaizumi, M., Ohtsuka, T., Matsushima, S., Akimoto, Y., Nishiwaki, C., Nakamichi, Y., Kikuta, T., Nagai, S., Kawakami, H., Watanabe, T., Nagamatsu, S
  • Journal Title: Mol Biol Cell 16 Pages: 3289-3300
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Rab27a mediates the tight docking of insulin granules onto the plasma membrane during glucose stimulation (2005)
  • Author(s): Kasai, K., Ohara-Imaizumi, M., Takahashi, N., Mizutani, S., Zhao, S., Kikuta, T., Kasai, H., Nagamatsu, S., Gomi, H., Izumi, T
  • Journal Title: J Clin Invest 115 Pages: 388-396
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Docking and fusion of insulin secretory granules in SUR1 knock out mouse beta-cells observed by total internal reflection fluorescence microscopy (2005)
  • Author(s): Kikuta, T., Ohara-Imaizumi, M., Nakazaki, M., Nishiwaki, C., Nakamichi, Y, Tei, C., Aguilar-Bryan, L., Bryan, J., Nagamatsu, S
  • Journal Title: FEBS Lett 579 Pages: 1602-1606
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Renin-angiotensin system modulates oxidative stress-induced endothelial cell apoptosis in rats (2005)
  • Author(s): Akishita, M., Nagai, K., Xi, H., Yu, W., Sudoh, N., Watanabe, T., Ohara-Imaizumi, M., Nagamatsu, S., Kozaki, K., Horiuchi, M., Toba, K
  • Journal Title: Hypertension 45 Pages: 1188-1193
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Imaging analysis reveals mechanistic differences between first and sec and ohase insulin exocytosis (2007)
  • Author(s): Ohara-Imaizumi, M(他4名)
  • Organizer: International Symposium on Membrane Traffic
  • Place of Presentation: Awaji,Japan
  • Year and Date: 20071127-29
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Imaging analysis reveals mechanistic differences between first and second phase insulin exocytosis (2007)
  • Author(s): Ohara-Imaizumi, M., Fujiwara, T., Akagawa, K., Nagamatsu, S
  • Organizer: International Symposium on Membrane Traffic
  • Place of Presentation: Awaji, Japan
  • Year and Date: 20071127-29
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Mechanism of insulin granule docking and exocytosis (2007)
  • Author(s): Nagamatsu, S., Ohara-Imaizumi, M.
  • Organizer: EASD islet study group symposium 2007
  • Place of Presentation: Brussels,Belgium
  • Year and Date: 20070921-23
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Mechanism of insulin granule docking and exocytosis, EASD islet study group symposium 2007 (2007)
  • Author(s): Nagamatsu, S., Ohara-Imaizumi, M
  • Organizer: Brussels
  • Place of Presentation: Belgium
  • Year and Date: 20070921-23
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Analysis of Newcomer Insulin Secretory Granule Movement by Newl y Developed Technique, Variable TIRF Real-Time Imaging System (2006)
  • Author(s): Ohara-Imaizumi M, , Nagamatsu S.(他4名)
  • Organizer: 66th Scientific Session of American Diabetes As sociation
  • Place of Presentation: Washinton.DC USA
  • Year and Date: 20060609-13
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Adiponectin Stimulates Insulin Secretion In Vitro and In Vivo (2006)
  • Author(s): Hashimoto S., Ohara-Imaizumi M., (他9名)
  • Organizer: 66th Scientific Session of American Diabetes As sociation
  • Place of Presentation: Washinton.DC USA
  • Year and Date: 20060609-13
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] PKCδ Plays an Essential Role in Glucose Stimulated Insulin Secretion Concurrent with the Enhanced Phosohorvlation of Munc18-1 (2006)
  • Author(s): Uchida T., Ohara-Imaizumi M., (他12名)
  • Organizer: 66th Scientific Session of American Diabetes Association
  • Place of Presentation: Washinton.DC USA
  • Year and Date: 20060609-13
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Analysis of Newcomer Insulin Secretory Granule Movement by Newly Developed Technique (2006)
  • Author(s): Ohara-Imaizumi, M., Nakamichi, Y., Wakazono, Y., Sakurai, T., Terakawa, S., Nagamatsu, S
  • Organizer: Variable TIRF Real-Time Imaging System 66th Scientific Session of American Diabetes Association
  • Place of Presentation: Washinton. DC
  • Year and Date: 20060609-13
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Adiponectin Stimulates Insulin Secretion In Vitro and In Vivo (2006)
  • Author(s): Hashimoto, S., Okamoto, M., Kubota, N., Ohara-Imaizumi, M, Eto, K., Takamoto, I., Yamaguchi, T., Ueki, K., Terauchi, Y., Nagamatsu, S., Kadowaki, T
  • Organizer: 66th Scientific Session of American Diabetes Association
  • Place of Presentation: Washinton. DC
  • Year and Date: 20060609-13
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] PKC Δ Plays an Essential Role in Glucose Stimulated Insulin Secretion Concurrent with the Enhanced Phosphorylation of Munc18-1 (2006)
  • Author(s): Uchida, T., Iwashita, N., Ohara-Imaizumi, M, Nagai, S., Choi, J B., Tamura, Y., Nomiyama, T., Ogihara, T., Tada, N., Tanaka, Y., Kawamori, R., Nagamatsu, S., Nakayama, K., Watada, H
  • Organizer: 66th Scientific Session of American Diabetes Association
  • Place of Presentation: Washinton. DC
  • Year and Date: 20060609-13
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] ELKS,a protein structurally related to the active zone associated protein CAST, is expressed in pancreatic beta cells and functions in insul in exocytosis (2005)
  • Author(s): Ohara-Imaizumi M, , Nagamatsu S.(他9名)
  • Organizer: 41st Annual Meeting of the EASD
  • Place of Presentation: Athens,Greece
  • Year and Date: 20050910-15
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] ELKS, a protein structurally related to the active zone associated protein CAST (2005)
  • Author(s): Ohara-Imaizumi, M, Ohtsuka, T, Matsushima, S, Akimoto, Y, Nishiwaki, C, Nakamichi, Y, Kikuta, T, Nagai, S, Kawakami, H, Watanabe, T, Nagamatsu, S
  • Organizer: is expressed in pancreatic beta cells and functions in insulin exocytosis 41st Annual Meeting of the EASD
  • Place of Presentation: Athens, Greece
  • Year and Date: 20050910-15
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] TIRF imaging analysis of the dynamic motion of insulin granules by antidiabetic agents (2005)
  • Author(s): Shinya Nagamatsu, Ohara-Imaiz umi M.
  • Organizer: 1st Scientific Meeting of the Asia-Pacific Diabetes and Obesity Study Group
  • Place of Presentation: Kobe
  • Year and Date: 20050820-21
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Visualized action of hypoglycemic agents,glinide and sulphonylurea,on the dynamic motion of insulin secretory granules analyzed by TIR F imaeine (2005)
  • Author(s): Ohara-Imaizumi M., Nagamatsu S.(他3名)
  • Organizer: 65th Scientific Session of American Diabetes As sociation
  • Place of Presentation: San Diego Jun.USA
  • Year and Date: 20050610-14
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Rab3 and SNAP25 in Islets are increased in insulin-resistant Irs2-/-mice and markedly decreased in.severely-hyperglycemic Irs2-/- mice on a high fat diet (2005)
  • Author(s): Suzuki R., Ohara-Imaizumi M., Nagamatsu S.(他8名)
  • Organizer: 65th Scientific Session of American Diabetes As sociation
  • Place of Presentation: San Diego Jun.USA
  • Year and Date: 20050610-14
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Visualized action of hypoglycemic agents, glinide and sulphonylurea, on the dynamic motion of insulin secretory granules analyzed by TIRF imaging (2005)
  • Author(s): Ohara-Imaizumi, M, Kikuta, T, Nishiwaki, C, Nakamichi, Y, Nagamatsu, S
  • Organizer: 65th Scientific Session of American Diabetes Association
  • Place of Presentation: San Diego
  • Year and Date: 20050610-14
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Rab3 and SNAP25 in Islets are increased in insulin-resistant Irs2-/- mice and markedly decreased in severely-hyperglycemic Irs2-/- mice on a high fat diet (2005)
  • Author(s): Suzuki, R, Tobe, K, Ohsugi, M, Sakamoto, K, Kamei, N, Nemoto, S, Ohara-Imaizumi, M, Kubota, N, Terauchi, Y, Nagamastu, S, Kadowaki, T
  • Organizer: 65th Scientific Session of American Diabetes Association
  • Place of Presentation: San Diego
  • Year and Date: 20050610-14
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] Mechanism of insulin exocytosis analyzed by imaging techniques.in Pancreatic Beta Cell in Health and Disease (2007)
  • Author(s): Nagamatsu S., Ohara-Imaizumi M.
  • Total Pages: 17
  • Publisher: Springer Verlag, Berlin Germany
  • Description: 「研究成果報告書概要(和文)」より