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Analysis of glucagon receptor by a novel functional cloning technique

Research Project

Project/Area Number05807087
Research Category

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

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

Principal Investigator

TSUKADA Toshihiko  Kyoto University Faculty of Medicine Department of Radiation Genetics, Instructor, 医学部, 助手 (10207334)

Project Period (FY) 1993 – 1994
Project Status Completed(Fiscal Year 1994)
Budget Amount *help
¥1,900,000 (Direct Cost : ¥1,900,000)
Fiscal Year 1994 : ¥500,000 (Direct Cost : ¥500,000)
Fiscal Year 1993 : ¥1,400,000 (Direct Cost : ¥1,400,000)
Keywordsbeta-Galactosidase / Menbrane Receptor / Vasoactive Intestinal Peptide / Proopiomelanocoertin / Gene Expression / Cyclic AMP / AtT-20 / PC12 / CAMP / POMC
Research Abstract

1.A chimeric gene containing the 5'-flanking region of the human proopiomelanocortin gene and E.coli lacZ gene was stably introduced into various cell lines including ACTH-producing mouse AtT-20 cells. The chimeric gene was expressed in AtT-20 cells in a cell-specific manner. A cell-spcific enhancer of the proopiomelanocortin gene was identified.
2.PC12-VG cells, which are PC12 rat pheochromocytoma cells stably transformed with a chimeric gene containing the 5'-flanking region of the human vasoactive intestinal peptide (VIP) gene, was developed. PC12-VG cells had minimal beta-galactosidase activities but showed high beta-galactosidase activity after treatment with reagents that activate protein kinase A.The PC12-VG cells were further transfected with a plasmid expressing the human luteinizing hormone receptor and stimulated with human chorionic gonadotropin showed high beta-galactosidase activity whereas no beta-galactosidase activity was detected in neither non-transfected nor unstimulated PC12-VG cells. Thus, PC12-VG cells were shown to be useful to detect exogenously introduced cDNA of membrane receptors that are positively coupled with adenylate cyclase.
3.VIP gene expression in PC12 cells were analyzed. Forskolin, an adenylate cyckase activating agent and TPA increased VIP mRNA content in PC12 cells as well as VIP release from PC12 cells. It was also shown that VIP induced VIP mRNA in PC12 cells.

Report

(3results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report

Research Products

(16results)

All Other

All Publications

  • [Publications] 中井 義勝: "Regulation of gene expression of pituitary hormones by hypophsiotropic hormones" Current Topics in Neuroendocrinology. 97-143 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 福島 光夫: "Pro-opiomelanocortin messenger RNA levels in anterior pituitaries of female Wistar fatty rats" The International Journal of Obesity. 17. 337-341 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 村上 典彦: "Bacterial lipopolysaccharide-induced expression of interleukin-6 messenger ribonucleic acid in the rat hypothalamus, pituitary, adrenal gland, and spleen" Endocrinology. 133. 2574-2578 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 塚田 俊彦: "Functional analysis of the cell-specific enhancer in the human pro-opiomelanocortin gene by β-galactosidasehistochemical staining" DNA and Cell Biology. 13. 755-762 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 塚田 俊彦: "An androgen receptor mutation causing androgen resistance in undervirilized male syndrome" The Journal of Clinical Endocrinology and Metabolism. 79. 1202-1207 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 塚田 俊彦: "Vasoactive intestinal peptide gene expression in rat pheochromocytoma cell line PC12" Molecular and Cellular Endocrinology. 107. 231-239 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yoshikatsu Nakai: "Regulation of gene expression of pituitary hormones by hypophysiotropic hormones" Current Topics in Neuroendocrinology. 97-143 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mitsuo Fukushima: "Pro-opiomelanocortin messenger RNA levels in anterior pituitaries of female Wistar fatty rats" The International Journal of Obesity. 17. 337-341 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Norihiko Murakami: "Bacterial lipopolysaccharide-induced expression of interleukin-6 messenger ribonucleic acid in the rat hypothalamus, pituitary, adrenal gland, and spleen" Endocrinology. 133. 2574-2578 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Toshihiko Tsukada: "Functinal analysis of the cell-specific enhancer in the human pro-opiomelanocortin gene by beta-galactosidase histochemical staining" DNA and Cell Biology. 13. 755-762 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Toshihiko Tsukada: "An androgen receptor mutation causing androgen resistance in undervirilized male syndrome" The Journal of Chinical Endocrinology and Metabolism. 79. 1202-1207 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Toshihiko Tsukada: "Vasoactive intestinal peptide gene expression in rat pheochromocytoma cell line PC12" Molecular and Cellular Endocrinology. 107. 231-239 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 塚田俊彦: "Functional analysis of the cell-specific enhancer in the human proopiomelanocortin gene by β-galactosidase histochemical staining" DNA and Cell Biology. 13. 755-762 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 塚田俊彦: "An androgen receptor mutation causing androgen resistance in undervirilized male syndrome" The Journal of Clinical Endocrinology and Metabolism. 79. 1202-1207 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 塚田俊彦: "Vasoactive intestinal peptide gene expression in rat pheochromocytoma cell line PC-12" Molecular and Cellular Endocrinology. 107. 231-239 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 塚田俊彦: "Functional analysis of the cell-specific enhancer in the human proopiomelanocortin gene by β-galactosidase histochemical staining" DNA and Cell Biology. (in press). (1994)

    • Related Report
      1993 Annual Research Report

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Published : 1993-04-01   Modified : 2016-04-21  

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