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PURIFICATION,CHARACTERIZATION AND SIGNAL TRANSDUCTION OF RETINAL CHOLINERGIC NEURON-NEUROTROPHIC FACTOR DERIVED FROM CULTURED HIPPOCAMPAL NEURONAL CELLS

Research Project

Project/Area Number 06671782
Research Category

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

Allocation TypeSingle-year Grants
Research Field Ophthalmology
Research InstitutionKansai Medical University

Principal Investigator

ITAGAKI Takashi  Kansai Medical University, Faculty of Medicine, Assistant Professor, 医学部, 講師 (30140255)

Co-Investigator(Kenkyū-buntansha) YASUKURA Tohru  Kansai Medical University, Faculty of Medicine, Reseach Associate, 医学部, 助手 (80257956)
OMORI Kyoko  Kansai Medical University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (90152256)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1994: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsHippocampus / Neurotrophic factor / choline acetyltransferase / Amacrin cells / Peptide / 神経栄養因子 / ペプチド / アマクリン細胞 / 脳海馬初代培養 / 網膜 / 細胞内情報伝達機構 / Neurdropluc factor / Chdineacityltrausferase / Amacnin cell(retina)
Research Abstract

A neurotrophic factor which supports the development of cholinergic retinal neuron (amacrine cells) was purified from media conditioned by a primary culture of embryonic hippocampal neurons.
Retinal choline acetyltransferase (ChAT), which is located exclusively in amacrine cells, served as a marker for the development of retinal cholinergic neurons. In serum-free control culture, retinal neuron from 17-day-old rat fetus displayd a little increase in the enzyme activity and low proportion of neurite-bearing cells (15-20%) within 7 days. The conditioned media, when added to the retinal neuron culture, dose-dependently increased ChAT activity and the number of neurite-bearing cells (40-60%), the maximum ChAT activity being - 6-fold higher than that in the control. The fraction with these stimulatory activities was purified by Sephadex G-15 column chromatography and two times reverse-phase (C18) HPLC.The final fraction showed - 3,000-fold higher purification as compared with that in the Sephadex G-15 fraction. Gas-phase protein sequencing analysis of the final fraction yielded a peptide sequence : Tyr-Leu-Leu-Pro-Ala-Gln-Val-Asn-Ile-Asp. A synthetic peptide with this sequence dose-dependently simulated ChAT activity in retinal cell culture and dissociated cell culture of the septal nucleus. The peptied also showed similar ChAT-stimulating activity in the dissociated cell culture of the septal nucleus. This peptide increased the phosphotyrosine residues of 180-200 kDa protein and the c-Fos synthesis in retinal culture, suggesting that the tyrosine kinase activation is involved in the response to this peptide. These findings suggest that the developing hippocampal neurons produce a neurotrophic peptide which stimulates the development of cholinergic neurons.
Thus hippocampal neurons appear to produce a neurotrophic peptide that promotes the development of cholinergic neurons amacrine cells.

Report

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

    (7 results)

All Other

All Publications (7 results)

  • [Publications] 保倉 透、他: "Purifition and Charactarization of New trofic Factor for retinal Cholinengic Neurcens Derined from Cultured Hippocampal Neuions" Jpn J.Pharomacol. 65. 153-162 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 大森京子: "Retinal Legeneration and Reguration" Kugler Pub.Ansterdem/New York(proceedings), 149-153 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tohru Yasukura, Kyoko Omori, Toshiko Mikami, Masafumi Inoue, Masanobu Uyama and Chiyoko Inagaki: "Purification and Characterization of Neurotrophic Factor for Retinal Cholinergic Neurons Derived from Cultured Hippocampal Neurons." Jap. J.Pharmacol. 65. 153-162 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kyoko Omori, Tohru Yasukura, Takashi Itagaki, and Chiyoko Inagaki.: Signal transduction events mediated by the Hippocampal neurotrophic factor for cholinergic retinal neurons in culture. Kugler Pub. Amsterdam/New York Proceedings of an International Symposium., 149-153 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kyoko Omori: "SIGNAL TRANSDUCTION EVENTS MEDIATED BY THE HIPPOCAMPAL NEUROTROPHIC FACTOR FOR CHOLINERGIC RETINAL NEUROSIN CULTURE" Retinal Degeneration and Regeueration, Proceedings of International Symposium. 1(印刷中). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Tohru YASUKURA: "Purification and Characterization of Neurotrophic Factor for Retinal Cholinergic Neurons Derived from Cultured Hippocampal Neurons" JPN J. Pharmacol. 65. 153-162 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 保倉 透,他: "Purification and Characterization of Neurotrphic Factor for Retirul Cldivergic Neiuon Denined from Cultiued Hippocampal Neuroris." Jpn.J.Pharmacol.65,. 65. 153-162 (1994)

    • Related Report
      1994 Annual Research Report

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

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