| Project/Area Number |
09480219
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurochemistry/Neuropharmacology
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| Research Institution | Gunma University |
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Principal Investigator |
SHIRAO Tomoaki Gunma Univ. Shc. Med. Dept. of Neurobiology and Behavior, Professor, 医学部, 教授 (20171043)
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| Co-Investigator(Kenkyū-buntansha) |
SAITOH Yoshiko Gunma Univ. Shc. Med. Dept. of Neurobiology and Behavior, Research Associate, 医学部, 助手 (70241263)
HAYASHI Kensuke Gunma Univ. Shc. Med. Dept. of Neurobiology and Behavior, Research Associate, 医学部, 助手 (50218567)
SEKINO Yuko Gunma Univ. Shc. Med. Dept. of Neurobiology and Behavior, Assist. Professor, 医学部, 講師 (70138866)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥13,100,000 (Direct Cost: ¥13,100,000)
Fiscal Year 1999: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥7,800,000 (Direct Cost: ¥7,800,000)
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| Keywords | dendritic spine / cytoskeleton / drebrin / actin / development / aging / synaptic maturation / 神経発達 / 神経培養 / 神経細胞 / 樹状突起 / スパイン / シナプス / グルタミン酸受容体 / 海馬 / アクチンフィラメント / 遺伝子導入 |
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| Research Abstract |
(A) Analysis of spine cytoskeletons
We prepare the cytoskeletal protein complex in the dendritic spine using the bead bound to anti-drebrin monoclonal antibody M2F6. The prepared sample contained actin, myosin I, myosin II, myosin V and gelsolin in addition to drebrin itself. Further, there are many other unidentified proteins in that sample ; therefore, we raised monoclonal antibodies using this sample as an immunogen. As a consequence we raised anti-myosin antibody and anti-CaMKII antibody. The anti-myosin antibody specifically recognized non-muscle myosin II heavy chan, which is present in cell soma and dendritic shaft in addition to dendritic spine. This indicates that spine cytoskelton is characterized by actin binding proteins but not by myosins.
(B) The change of spine cytoskeletons during development and aging.
In the developing brain, drebrin E is observed in the migrating neurons and within the growing axons and dendrite. In the adult brain, drebrin A is localized in the dendrit
… More
ic spines. In this study we investigated when drebrin disappeared from cell soma and enriched in dendritic spines during development. Immunohistochemical staining of the cerebrum and cerebellum using anti-drebrin monoclonal antibody (MBL, Japan) demonstrated that drebrin immunoreactivity was observed in the cell soma and dendrites from postnatal 0 day to 10 day, although drebrin expression as a whole was decreased according to the postnatal days. However, in 14-day-old rat, drebrin immunoreactivity was only observed as dot-like pattern in neuropil. Drebrin immunoreactivity was hardly observed within the cell soma and dendrites. These data indicates that drebrin subcellular localization was changed synchronously around postnatal 12 days, indicating that the final maturation of neuronal cytoskeleton may occurred at postnatal 12 days. Application of 100 μM glutamate weakened drebrin immunoreactivity at dendritic spines. Transfection experiments demonstrated that overexpression of drebrin A in neuron resulted the elongation of the spine length. These data indicate that developmental changes in the distribution of drebrin from dendritic shafts into dendritic spines may be related to the maturation of synaptic function. Less
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