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Distribution of drebrin containing synapses in the brain

含有突触的drebrin在大脑中的分布

基本信息

批准号：
10044237
负责人：
SHIRAO Tomoaki
金额：
$ 1.34万
依托单位：
Gunma University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1998
资助国家：
日本
起止时间：
1998 至无数据
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10044237/
关键词：
drebrin spine neuron cyto keleton actin synapse development aging

项目摘要

1. We have systematically analyzed the Alzheimer's brain using biochemical and histol6gical technique. Dr.Hatanpaa demonstrated that drebrin expression was downregulated more specifically than other synaptic functional proteins. He also showed that drebrin expression decreased in parallel with normal human aging. Drs.Shirao and Sekino analyzed distribution of drebrin containing synapses in the rat brain during development. Wistar rats of various developmental stages from postnatal day 0 to day 20 were transcardially perfused with 3.7% formalin and the brain was removed, postfixed, and their cryostat section was immunostained with ABC method or with immunofluorescence. Before postnatal by 10 drebrin remain to be observed in cell body, axons and dendrites. In contrast at postnatal day 14 drebrin was localized in the dendritic spines, and was not observed in cell soma.2. Then we examined whether this developmental change in drebrin distribution was observed in primary cultured neurons or not. At 7 days in culture, drebrin was observed in neurites, and its staining pattern is continuous, In the cell soma drebrin was concentrated in cortical cytoplasm. Although at this developmental stage spine formation was started, drebrin clusters were not yet observed. At 14 days in culture drebrin staining pattern was most discontinuous and patchy although thin neurites was continuously immunostained. At 21 days in culture all drebrin-staining pattern was patchy. Double immunostaining with synaptophysin and drebrin antibodies demonstrated that drebrin is accumulated in the dendritic spines. Taken together it was demonstrated that drebrin accumulation was occurred in parallel with the maturation of synaptic function, and was indicated that this accumulation was regulated by neuronal activity.

1.应用生化和组织学技术对阿尔茨海默病脑进行了系统的分析。Hatanpaa博士证明，与其他突触功能蛋白相比，Drebrin的表达更具特异性地下调。他还表明，随着人类的正常衰老，Drebrin的表达也随之减少。Shirao博士和Sekino博士分析了发育过程中大鼠大脑中含有Drebrin的突触的分布情况。取出生后第0天至第20天不同发育阶段的Wistar大鼠，经心内注入3.7%福尔马林，取脑后固定，冰冻切片用ABC法或免疫荧光染色。出生后10天以前，在细胞体、轴突和树突中仍可观察到DREBRIN。结果：1.生后14天，DREBRIN定位于树突棘，细胞体中未见DREBRIN。然后，我们检查了在原代培养的神经元中是否观察到了这种DREBRIN分布的发育变化。培养7d时，神经元突起内可见Drebrin，染色呈连续性，胞体中Drebrin主要集中在皮质细胞质中。虽然在这个发育阶段已经开始形成脊椎，但还没有观察到树突簇状结构。培养14d时，虽然有细小的神经突起连续免疫染色，但Drebrin染色模式最不连续，呈斑片状。培养到第21天时，所有的Drebrin染色模式都是斑片状的。突触素和Drebrin抗体双重免疫染色显示，Drebrin聚集在树突棘中。综上所述，突触功能成熟的同时也出现了脑干蛋白的蓄积，并提示这种蓄积受神经元活动的调节。