| Project/Area Number |
01570026
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
神経解剖学
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| Research Institution | Chiba University, School of Medicine |
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Principal Investigator |
KADOTA Tomoko Chiba University, School of Medicine, Department of Anatomy, Associate Professor, 医学部, 助教授 (00089864)
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| Co-Investigator(Kenkyū-buntansha) |
TAGAWA Masatoshi Chiba University, School of Medicine, Center for Neurobiology and Molecular Immu, 神経センター・国府台病院, 厚生技官 (20171572)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1990: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1989: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | synaptic vesicle / synaptophysin / 82K protein / monoclonal antibody / postsynaptic density / immunohistochemistry / immunoelectron microscopy / 82k蛋白 / シナプトフィシン / 神経細胞 / シナプス複合体 |
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| Research Abstract |
1. Postsynaptic density specific protein, 82K protein
A fraction of synaptic junctional complex (SJC) was prepared from rat brain synaptosomes and served as an antigen material to produce monoclonal antibodies for examining the component proteins of the SJC. An antibody, SJ-8, was obtained, which recognized a protein with a molecular weight of 82,000 dalton (82K protein) in the SJC preparation in immunoblotting analysis. The immunohistochemical localization of the 82K protein was studied with the rat cerebellum. The antibody SJ-8 labeled the peripheral area of the Purkinje and granule cells. Small punctate areas were also stained in the molecular layer with SJ-8. Intracellular localization of the protein was examined with the rat brain synaptosomes. Immunoelectron microscopy demonstrated that SJ-8 strongly labeled the postsynaptic density and also a fibrous network spreading out of it. However, the antibody did not label the pre- and postsynaptic membrane.
2. Expression of synaptophysin
… More
in the rat cerebellum
Expression and intracellular localization of synaptophysin was examined in cerebellar cortex of the perinatal rat. Rats, 1-7 days after birth, were fixed by perfusion via the heart with 4% paraformaldehyde. The cerebellar cortex was dissected out and cut into 40 mum thick slices with a vibratome. After blocking with 1% bovine serum albumin, the slices were incubated with the antibody against synaptophysin. Then, slices were reacted with FITC- conjugated anti-mouse IgG and examined with a light microscopy equipped with epifluorescence. Specimens from perinatal rats did not show any positive reactions. Several positive spots appeared around the Purkinje cells and granule cells three days after birth. Small punctates of reaction were also observed in the molecular layer. These positive reactions significantly increased 7 days after birth. The reaction pattern in the cerebellar cortex after 2 weeks was similar to that of adult rats. The development of positive reaction may involve the synaptogenesis in the cerebellar cortex. Synaptophysin might be transported from the perikaryon to the nerve ending via the axon before the synaptogenesis. The transporting system of the protein is immunocytochemically examined. Less
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