Development and Aging of Neuronal Synapse

神经元突触的发育和衰老

基本信息

批准号：
09480219
负责人：
SHIRAO Tomoaki
金额：
$ 8.38万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

(A) Analysis of spine cytoskeletonsWe 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

（a）使用珠子链球链球菌I II，肌球蛋白II，肌球蛋白V和凝胶蛋白，对树突状脊柱的细胞骨架蛋白质复合物进行分析。在该样品中，未识别的蛋白会提出抗肌球蛋白抗体和抗CAMKII抗体蛋白质而不是肌动物，在迁移的神经元和轴突中观察到了蛋白质使用抗杀伤蛋白单克隆抗体的小脑，在细胞体内观察到drebrin免疫反应性，而在14天大的大鼠中，在产后0天到10天，在产后0天到10天。仅在细胞体内观察到DREBRIN免疫反应性的DOT样模式。在树突状的旋转中，脊柱长度的伸长表明，树突轴的发育变化可能会变化