Diversity of cholinergic neurons in the human central nervous system

人类中枢神经系统胆碱能神经元的多样性

基本信息

批准号：
11670171
负责人：
YOSHIO Oda
金额：
$ 2.11万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

Choline acetyltransferase (ChAT), an enzyme for biosynthesis of acetylcholine, and vesicular acetylcholine transporter (VAChT), a molecule for the translocation of cytoplasmic acetylcholine into synaptic vesicles, are known to be good markers for cholinergic neurons. In order to investigate whether cholinergic neurons are diverse with relation to localization and function, immunohistochemical and in situ hybridization studies were performed on the human central nervous by using specific antibodies and RNA probes for human ChAT and VAChT.In this study, an antibody to human recombinant VAChT protein was developed, which was applicable to paraffin-embedded tissue sections. The distributions of ChAT mRNA, ChAT protein, VAChT mRNA and VAChT protein were similar, but not exactly the same in the human central nervous system. In addition, cholinergic projection neurons tended to express these molecules more than cholinergic local circuit neurons, which might be related to the different functional levels between the two types of cholinergic neurons. Southern blot analysis following reverse transcription- polymerase chain reaction revealed that the caudate nucleus contained two R-type mRNAs, but not N-or M-type mRNA.Our previous study demonstrated that in the human spinal cord M-type mRNA is much more abundant than R-type mRNA.These results suggest that transcription mechanisms of ChAT are different in cholinergic neurons between the caudate nucleus and spinal cord.

胆碱乙酰转移酶（ChAT）是一种用于乙酰胆碱生物合成的酶，而囊泡乙酰胆碱转运蛋白（VAChT）是一种将细胞质乙酰胆碱转运到突触小泡的分子，已知它们是胆碱能神经元的良好标记物。为了研究胆碱能神经元在定位和功能方面是否存在多样性，通过使用针对人 ChAT 和 VAChT 的特异性抗体和 RNA 探针，对人中枢神经进行了免疫组织化学和原位杂交研究。开发了重组VAChT蛋白，适用于石蜡包埋组织切片。 ChAT mRNA、ChAT蛋白、VAChT mRNA和VAChT蛋白在人中枢神经系统中的分布相似，但不完全相同。此外，胆碱能投射神经元比胆碱能局部回路神经元更倾向于表达这些分子，这可能与两类胆碱能神经元之间的功能水平不同有关。逆转录聚合酶链反应后的 Southern blot 分析显示，尾状核含有两个 R 型 mRNA，但不含有 N 型或 M 型 mRNA。我们之前的研究表明，在人类脊髓中，M 型 mRNA 的含量要丰富得多这些结果表明ChAT的转录机制在尾状核和脊髓之间的胆碱能神经元中是不同的。