1993 Fiscal Year Final Research Report Summary
Cell-site specific synapse formation in culture
| Project/Area Number |
04454134
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurophysiology and muscle physiology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
HIRANO Tomoo Kyoto University, Department of Physiology, Associate Professor, 医学部, 助教授 (50181178)
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| Project Period (FY) |
1992 – 1993
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| Keywords | Synapse formation / Cerebellum / Culture / Purkinje cell / Granule cell / Basket cell / Stellate cell / Fluorescent staining |
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| 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.
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