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2003 Fiscal Year Final Research Report Summary

A study on the Intracellular dynamics of insulin granules using a phogrin-GFP fused protein

Research Project

Project/Area Number 13470006
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General anatomy (including Histology/Embryology)
Research InstitutionFujita Health University

Principal Investigator

SENDA Takao  Fujita Health University, School of Medicin, Professor, 医学部, 教授 (10187875)

Co-Investigator(Kenkyū-buntansha) HASEGAWA Yoshimi  Fujita Health University, School of Medicine, Research Associate, 医学部, 助手 (40288494)
HIDA Takehiko  Fujita Health University, School of Medicine, Associate Professor, 医学部, 助教授 (20097736)
NIKI Ichiro  Oita University, School of Medicine, Professor, 医学部, 教授 (10262908)
Project Period (FY) 2001 – 2003
Keywordsinsulin secretion / MIN6 cells / pancreatic B cells / phogrin / glucose / calmodulin / apoptosis / caveolin-1
Research Abstract

1.We constructed an expression vector in which phogrin is bound to the C-terminus of GFP. Using the vector, we successfully expressed. GFP-phogxin in MIN6 cells.
2.We got several new findings concerning regulation mechanisms of the insulin granule traffic.
(1)Glucose accelerated the insulin granule traffic in MIN6 cells. The calmodulin-inhibitor W-7 inhibited the glucose-induced acceleration of the granule traffic, and myosin light chain kinase inhibited the glucose-induced acceleration of the granule traffic.
(2)The blockers to Ca^<2+> channels (nifedipine and nitrendipine) did not inhibit the glucose-induced acceleration of the granule traffic.
(3)The K_<ATP>-opener (diazoxide) did not inhibit the glucose-induced acceleration of the granule traffic.
(4)The glucose stimulation increased phosphatized myosin light chain in MIN6 cells.
3.We investigated the mechanism of B cell loss in transgenic mice with elevated levels of B cell calmodulin.
(1)The transgenic mice experienced a sudden rise in blood glucose levels, which was associated with developed of severe hypoinsulinemia and loss of B cells from the islets.
(2)Ultrastructural analysis revealed that compromised granule formation and apoptotic changes in the transgenic B cells preceded the onset of hyperglycemia.
(3)Intraperitoneal injection of tolbutamide, an antidiabetic sulfonylurea, decreased blood glucose levels but increased the number of apoptotic B cells.
(4)Injection of the mice with N^ω-nitro-L-arginine methyl ester, which inhibits nitric oxide synthase activity, prevented hyperglycemia and lessened the changes in number and size of B cells.
4.We found that caveolin-1 was expressed in MIN6 cells, and colocalized with γ-tubulin in microtubule-organizing center. The signal of caveolin-1 decreased after glucose stimulation.

  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Nakanishi M et al.: "Nuclear targeting of DNA."Eur J Pharmaceutical Sci. 13. 17-24 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Niwa T et al.: "Characterization of secretory and morphologic properties of primary cultured endocrine cells from porcine pancreata."Pancreas. 22. 135-140 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Eguchi A et al.: "Protein transduction domain of HIV-1 Tat protein promotes efficient delivery of DNA into mammalian cells."J Biol Chem. 276. 26204-26210 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Niki I et al.: "Regulation of insulin release at pre-exocytotic stages of the secretory process."Curr Med Chem-Imun Endoc& Metab Agents. 2. 219-231 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Fan QW et al.: "Cholesterol-dependent modulation of dendrite outgrowth and microtubule stability in cultured neurons."J Neurochem. 80. 178-190 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tsukamoto Y et al.: "Arpp, a new homologue of Carp, is preferentially expressed in type 1 skeletal muscle fibers and is markedly induced by denervation."Lab Invest. 82. 645-655 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yu W et al.: "Calmodulin overexpression causes Ca^<2+>-dependent apoptosis of pancreatic β cells, which can be prevented by inhibition of nitric oxide synthase."Lab Invest. 82. 1229-1239 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Akuta T et al.: "Enhancement of phage-mediated gene transfer by nuclear localization signal."Biochem Biophys Res Commun. 297. 779-786 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakanishi M et al.: "Basic Peptides as Functional Components of Non-viral Gene Transfer Vehicles."Current Protein and Peptide Science. 4. 141-150 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shimomura A, Nomura R, Senda T: "Lithium inhibits apoptosis of mouse neural progenitor cells."Neuroreport. 14. 1779-1782 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Niki I et al.: "Ca^<2+> influx does not trigger glucose-induced traffic of the insulin granules and alteration of their distribution."Exp Biol Med. 228. 1218-1226 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Senda T, Nomura R: "The expression of cytokeratin in hepatic stellate cells of the cod."Arch Histol Cytol. 99. 437-444 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakanishi M et al.: "Nuclear targeting of DNA."Eur J Pharmaceutical Sci. 13. 17-24 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Niwa T et al.: "Characterization of secretory and morphologic properties of primary cultured endocrine cells from porcine pancreata."Pancreas. 22. 135-140 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Eguchi A et al.: "Protein transduction domain of HIV-1 Tat protein promotes efficient delivery of DNA into mammalian cells."J Biol Chem. 276. 26204-26210 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Niki I et al.: "Regulation of insulin release at pre-exocytotic stages of the secretory process."Curr Med Chem -Imun Endoc & Metab Agents. 2. 219-231 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fan QW et al.: "Cholesterol-dependent modulation of dendrite outgrowth and microtubule stability in cultured neurons."J Neurochem. 80. 178-190 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tsukamoto Y et al.: "Arpp, a new homologue,of Carp, is preferentially expressed in type 1 skeletal muscle fibers and is markedly induced by denervation."Lab Invest. 82. 645-655 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yu W et al.: "Calmodulin overexpression causes Ca^<2+>-dependent apoptosis of pancreatic β cells, which can be prevented by inhibition of nitric oxide synthase."Lab Invest. 82. 1229-1239 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Akuta T et al.: "Enhancement of phage-mediated gene transfer by nuclear localization signal."Biochem Biophys Res Commun. 297. 779-786 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakanishi M et al.: "Basic peptides as functional components of non-viral gene transfer vehicles."Current Protein and Peptide Science. 4. 141-150 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shimomura A, Nomura R, Senda T: "Lithium inhibits apoptosis of mouse neural progenitor cells."Neuroreport. 14. 1779-1782 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Niki I et al.: "Ca^<2+> influx does not trigger glucose-induced traffic of the insulin granules and alteration of their distribution."Exp Biol Med. 228. 1218-1226 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Senda T, Nomura R: "The expression of cytokeratin in hepatic stellate cells of the cod."Arch Histol Cytol. 66. 437-444 (2003)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2005-04-19  

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