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1991 Fiscal Year Final Research Report Summary

Cellular Response to Damage of Axonal Transport in Cultured Retinal Cells

Research Project

Project/Area Number 01570982
Research Category

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

Allocation TypeSingle-year Grants
Research Field Ophthalmology
Research InstitutionKitasato University

Principal Investigator

WAKAKURA Masato  Kitasato Univ. Ophthalmology, Associate Professor, 医学部, 助教授 (50137931)

Co-Investigator(Kenkyū-buntansha) FUKUSHIMA Kazuya  Kitasato Univ. Ophthalmology Assistant, 医学部, 助手 (60199205)
YAMAMOTO Noboru  Kitasato Univ. Functional Morph. Professor, 看護学部, 教授 (10050543)
Project Period (FY) 1989 – 1991
KeywordsMuller cells / retinal neurons / axonal transport / neurotransmitters / aminoadipic acid / 4-aminopyridine / culture / glial fibrillary acidic protein
Research Abstract

A number of neurotransmitters elicit postsynaptic release through influx of extracellular calcium ion. Retinal neurons cultured from neonatal rat eyes, in which pure Muller cell culture was used as a feeder layer, expressed not only neurofilament peptide also various neurotransmitters. Using the cultured retinal neurons, cellular response to 4-aminopyridine(4-AP)was investigated. Free calcium in the neurons was labelled with Fura-2 AM and the rapid change in intracellular calcium ion[Ca^<2+>]_i was analyzed by a computerassisted fluorospectrometry. Rapid increase of [Ca^<2+>]_i was seen significantly following treatment beyond 4mM of 4-AP in all neurons. Since the response was not completely blocked during incubation with calcium-free EGTA containing medium, intracellular calcium mobilization system appeared also to be exerted.
Administration of excitatory amino acids may be toxic for retinal cells and block axonal transport in retinal neurons. Effects of aminoadipic acid(AAA)on culture … More d retinal cells were investigated by the above method of [Ca^<2+>]_i measurement. The D, L- and D-forms of AAA activated neurons at low concentrations and activated the Muller cells at higher concentrations. The D-form may act selectively on retinal neurons, suggesting that it may be an agonist of an excitatory amino acid receptor. The results indicate that the ability of AAA to elevate cytosolic [Ca^<2+>]_i depends upon the spectrospecificity of the AAA and on cell type. The prolonged effect of D, L-AAA on cultured retinal cells was also studied using morphological observation and immunofluorescence of glial fibrillary acidic, protein(GFAP). Vacuolations in Muller cells were found after 8.0 mM treatment. Reduction of neurons was evident after treatment beyond 1.6 mM. Administration of AAA was followed by accumulation of GFAP in Muller cells but never resulted in reduction of the cells or reduction of DNA content in the culture, suggesting strong. cellualr resistance. These results indicate AAA not to be highly toxic for Muller cells and to be rather toxic for retinal neurons. The results support that the cellular response to AAA depends upon Cellular constituent. Less

  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] 若倉 雅登: "細胞のストレス応答と眼" 日本眼科学会雑誌. 93. 511-523 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wakakura M: "Effects of 4-aminopyridine on intra cellular calcium mobilization in transmitter-containing retinal neurons in vitro." Invest Ophthalmol Vis Sci (ARVO abstr). 31. 335 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wakakura M et al: "Direct effect of DL-2-aminoadipic acid mediated by calcium ion cultured Muller cells." Doc Ophthalmol. 76. 109 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 若倉 雅登ほか: "培養網膜ニュ-ロンのカルシウム動員系について" 日本眼科学会誌. 94(増). 177 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 若倉 雅登: "視覚神経伝達物質ー培養細胞を用いた解析ー" 神経眼科. 7. 190-195 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wakakura M: "Rapid sncrease of in tracellular Ca^<2+> concentratin caused by aminoadipic acid isomer in Muller cells and neurons in vitro." Doc Ophthalmol.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wakakura M: "Cellular stress response in the eye." Nippon Ganka Gakkaizasshi. 93. 511-523 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wakakura M: "Effects of 4-aminopyridine on intracellular calcium mobilization in transmitter-containing retinal neurons in vitro." Invest Ophthalmol Vis Sci (ARVO abstract). 31. 335 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wakakura M, Yamamoto N: "Direct effect of DL-2-aminoadipic acid mediated by calcium ion on cultured Muller cells." Doc Ophthalmol. 76. 109 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wakakura M: "retinal transmitters : An approach using cultured retinal neurons." Shinkeiganka. 7. 190-195 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wakakura M, Yamamoto N: "Rapid increase of intracellular calcium ion concentration caused by aminoadipic acid isomers in Muller cells and neurons in vitro." Doc Ophthalmol.

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

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Published: 1993-03-16  

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