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1993 Fiscal Year Final Research Report Summary

Cell-site specific synapse formation in culture

Research Project

Project/Area Number 04454134
Research Category

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

Allocation TypeSingle-year Grants
Research Field Neurophysiology and muscle physiology
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

HIRANO Tomoo  Kyoto University, Department of Physiology, Associate Professor, 医学部, 助教授 (50181178)

Project Period (FY) 1992 – 1993
KeywordsSynapse formation / Cerebellum / Culture / Purkinje cell / Granule cell / Basket cell / Stellate cell / Fluorescent staining
Research Abstract

Spatial distribution of synapses on Purkinje cells formed in a dissociated cell culture of rat cerebellum, was studied by intracellular fluorescent stainings of pre- and postsynaptic neurons and by immunocytochemical staining of presynaptic terminals. A Purkinje cell was whole-cell voltage-clamped and filled with Texas red. Then, it was fixed and stained with an anti-synaptic vesicle monoclonal antibody using FITC.This staining revealed that a Purkinje cell receives synaptic inputs on both a soma and dendrites. Simultaneous whole-cell recordings were performed on a presynaptic small neuron and a Purkinje cell, and the property of synaptic transmission was determined. At the same time, the presynaptic neuron was filled with Lucifer yellow and the Purkinje cell was filled with Texas red. Axonal swellings presumed to be synaptic terminals of an excitatory granule cell, were exclusively localized along dendrites of a Purkinje cell, and presumed synaptic terminals of an inhibitory interneuron were found on both dendrites and a soma. Synapses were also observed with a scanning electron microscope. Antibodies were raised against one subtype of glutamate receptors, mGluR1, and distribution pattern of mGluR1 on a Purkinje cell was examined with immunocytochemical staining. Punctate labelings of mGluR1 were observed along dendrites of cultured Purkinje cells.mGluR1 were distributed more on dendrites than on a cell body. This distribution pattern of mGluR1 is consistent with that of excitatory glutamatergic synapses from granule cells. Thus, cell-site specific synapse formations and also cell-site specific distribution of the receptor have been established in a simple culture system.

  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Hirano,T and Kasono,K: "Spatial distribution of excitatory and inhibitory synapses on a Purkinje cell in a rat cerebellar culture" Journal of Neurophysiology. 70. 1316-1325 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirano,T.: "Cellular and molecular mechanism of synaptic depression in a cerebellar culture." Functional Neurology. 8(4). 25-26 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shigemoto,R.,Nakanishi,S.and Hirano,T.: "Antibodies against a metabotropic glutamate receptor,mGluR1,inhibit long-term depression in cerebellar culture." Neuroscience Research Suppl.18. S46 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kasono,K.and Hirano,T.: "Critical role of postsynaptic calcium in cerebellar long-term depression in culture." Neuroscience Research Suppl.18. S147 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 平野 丈夫: "小脳培養系におけるシナプスと可塑性" ブレインサイエンス最前線'94 講談社サイエンティフィック. 98-109 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirano, T.and Kasono, K.: "Spatial distribution of excitatory and inhibitory synapses on a Purkinje cell in a rat cerebellar culture." Journal of Neurophysiol.70. 1316-1325 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirano, T.: "Cellular and molecular mechanism of synaptic depression in a cerebellar culture." Functional Neurology. 8(4). 25-26 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shigemoto, R., Nakanishi, S.and Hirano, T.: "Antibodies against a metabotropic glutamate receptor, mGluR1, inhibit long-term depression in cerebellar culture." Neurosc.Res.Suppl.18. S46 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kasono, K.and Hirano, T.: "Critical role of postsynaptic calcium in cerebellar long-term depression in culture." Neurosci.Res.Suppl.18. S147 (1993)

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

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Published: 1995-03-27  

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