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1998 Fiscal Year Final Research Report Summary

Regulation of hypothalamic peptides by in vivo electroporation and its physiological effects

Research Project

Project/Area Number 09671063
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 内分泌・代謝学
Research InstitutionKagawa Medical University

Principal Investigator

SATO Makoto  Kagawa Medical University, 1st Dept Int Med, Research Assosiate, 医学部, 助手 (10215840)

Co-Investigator(Kenkyū-buntansha) TAKAHARA Jiro  Kagawa Medical University, 1st Dept Int Med, Professor, 医学部, 教授 (00033085)
Project Period (FY) 1997 – 1998
Keywordselectroporation / hypothalamus / oxytocin / vasopressin / beta-galactosidase / retrograde transport / rat / plasmid
Research Abstract

To investigate physiological functions of hypothalamic peptides, it is important to control their expression at transcription levels. In this study, we attempted to establish in vivo electroporation to transfer an exogenous gene into hypothalamic nucleus in the rat. An expression vector, pCMVbeta, was used to examine the efficiency of gene transfer. The plasmid was injected into the lateral cerebral ventricle in rats and a bipolar electrode was inserted outside the hypothalamic arcuate nucleus (ARC). A series of electric pulses were delivered with a standard square wave electroporator. After the electroporation, brains of the rats were perfused with 4 % paraformaldehyde and cryostat sections were cut. The beta -galactosidase activity was examined using X-Gal histochemical stain. Staining tissues that express beta -galactosidase with X-Gal turned blue. Expression of beta -galactosidase was found in the choroid plexus, paraventricular nucleus (PVN), and supraoptic nucleus (SON). Although the electrode was located closely arround the hypothalamic ARC, the expression of beta -galactosidase was not observed in the ARC.Plasmid was also not incorporated into the ARC despite of the microinjection above the nucleus. The choroid plexus, PVN and SON showed positive expression of beta -galactosidase after injection of the plasmid into the carotid artery. These results suggest that the in vivo electroporation method might have a limited use for introducing exogenous genes in to the hypothalamic nucelus. The neurons that express beta -galactosidase in the PVN and SON appear to be oxytocin or vasopression, because these neurons are mostly abundant in both nuclei. The unexpected observation that the plasmid was incorporated into the PVN and SON raises the possibility that exogenous DNA may reach to these hypothalamic nuclei through their retrograde axonal transport. Further studies are required to establish a new method of in vivo gene transfer by the retrograde DNA transport.

  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Michio Niimi: "Effect of continuous intravenous injection of interleukin-6 and pretreatment with cyclooxygenase inhibhitor on brain c-fos expression in the rat." Neuroenodocrinology. 66. 47-53 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Makoto Sato: "Gene regulation of growth hormone-releasing hormone and its receptor." EndocrJ. 44. 765-774 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shuji Matsubara: "Quantitative analysis of growth hormone(GH)pre-mRNA expression in cultured rat anterior pituitary cells by an intron-specific and competitive PCR method." Endocrinology. 138. 5075-5078 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ryoji Yokote: "Molecular cloning and gene expression of growth hormone-releasing peptide receptor in rat tissues." Peptides. 19. 15-20 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomoyo Ohyama: "Effects of adrenalectomy and glucocorticoid receptor antagonist,RU38486, on pituitary growth hormone-releasing hormone receptor gene expression in rats." Peptides. 19. 1063-1067 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Michio Niimi: "Effects of central and peripheral injection of leptin on food intake and on brain c-fos expression in the Otsuka Long-Evans Tokushima Fatty rat with hyperleptinemia." J Neuroendocrinol. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 佐藤誠: "医学大事典, インスリン低血糖試験など11項目(印刷中)" 医学書院,

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 高原二郎: "内科学教科書, 下垂体-標的器官の機能テストと評価(印刷中)" 黒川清・松澤祐次編集主幹, 文光堂,

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Michio Niimi: "Effect of continuous intravenous injection of interleukin-6 and pretreatment with cyclooxygenase i hibhitor on brain c-fos expression in the rat." Neuroenodocrinology. 66. 47-53 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto Sato: "Gene regulation of growth hormone-releasing hormone and its receptor." Endocr J. 44. 765-774 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shuji Matsubara: "Quantitative analysis of growth hormone (GH) pre-mRNA expression in cultured rat anterior pituitary cells by an intron-specific and competitive PCR method." Endocrinology. 138. 5075-5078 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ryoji Yokote: "Molecular cloning and gene expression of growth hormone-releasing peptide receptor in rat tissues." Peptides. 19. 15-20 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomoyo Ohyama: "Effects of adrenalectomy and glucocorticoid receptor antagonist RU38486, on pituitary growth hormone-releasing hormone receptor gene expression in rats." Peptides. 19. 1063-1067 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Michio Niimi: "Effects of central and peripheral injection of leptin on food intake and on brain c-fos expression in the Otsuka Long-Evans TokushimaFatty rat with hyperleptinemia." J Neuroendocrinol. (in press).

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

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Published: 1999-12-08  

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