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Mechanism of insulin action on pancreatic Alpha cells-gene analysis of insulin receptor-

Research Project

Project/Area Number 06671043
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 内分泌・代謝学
Research InstitutionKUMAMOTO UNIVERSITY

Principal Investigator

KISHIKAWA Hideki  Kumamoto University School of Medicine, Lecturer, 医学部, 講師 (30161441)

Co-Investigator(Kenkyū-buntansha) ARAKI Eiich  Kumamoto University School of Medicine, Research Associate, 医学部, 助手 (10253733)
矢野 智彦  熊本大学, 医学部・附属病院, 助手 (90166562)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsPancreatic Alpha cell / In-R1-G9 cells / alpha TC clone 6 / Insulin receptor / Glucagon secretion / In-R1-G9細胞 / αTC クローン6細胞 / グルカゴン分泌腫瘍細胞 / αTC clone6 細胞
Research Abstract

In pancreatic Alpha cells, the existence and function of insulin receptor has not been fully studied yet. In this study, to confirm the expression of functional insulin receotor in pancreatic Alpha cells, we perfomed 1)insulin receptor binding assay, 2)Northern blot analysis and RT-PCR (reverse transcription-polymerase chain reaction) amplification of insulin recetor mRNA,3)immunocytochemical staining, 4)biosynthetical labelling of insulin receptor protein using [^<35>S] methionine, 5)the analysis of insulin-stimulated autophosphorylation of the insulin receptor in glucagon secreting cell lines ; In-R1-G9 and alpha TC clone 6 cells.
The glucagon secretion decreased by the addition of insulin in both cells. The receptor binding studies using[125I-Tyr-A14] insulin revealed that both cells possessed the significant number of insulin receptors (In-R1-G9 : K_1=2.1x10^9mol/l, K_2=6.2x10_7mol/l, R_1=0.27x10^4, R_2=1.86x10^4sites/cell ; alpha TC clone 6 : K_1=2.1x10^9mol/l, K_2=7.3x10^7mol/l, R_1=0.27x10^4, R_2=1.95x10^4sites/cell). Northern blot analysis as well as RT-PCR amplification showed the mRNA specific for insulin receptor in both cells. By immunocytochemical staining using antiinsulin receptor a subunit antibody, positive immunostainings for insulin receptor were observed in both cells. [^<35>S] methionine labelling of both cells followed by immunoprecipitation using antiinsulin receptor antibody showed correct size of insulin receptor protein. The insulin receptor expressed in these cells underwent autophosphorylation by insulin stimulation.
It is concluded that functional insulin receptors are properly expressed in In-R1-G9 and alpha TC clone 6 cells.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (4 results)

All Other

All Publications (4 results)

  • [Publications] 木佐貫浩一,他: "Expression of insulin receptor on clonal pancreatic Alpha cells and its possible role for insulin-stimulated negative regulation of glucagon secretion" DIABETOIOGIA. 38. 422-429 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Kisanuki, et al: "Expression of insulin receptor on clonal pancreatic Alpha cells and its possible role for insulinstimulated negative regulation of glucagon secretion." Diabetologia. 38. 422-429 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 木佐貫浩一他: "Expression of insulin receptor on clonal pancreatic Alpha cells and its possible role for insulin-stimulated negative regulation of glucagon secretion" DIABETOIOGIA. 38. 422-429 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 木佐貫浩一: "グルカゴン分泌腫瘍細胞(In-R1-G9)におけるインスリン受容体の生化学的免疫組織学的,遺伝子工学的検討" PEPTIDE HORMONE IN PANCREAS. 13. 168-89 (1993)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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