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Immunohistochemical studies of dopamine receptor subtypes in schizophrenic brain by using anti-subtype specipfic peptide antibodies

使用抗亚型特异性肽抗体对精神分裂症脑中多巴胺受体亚型进行免疫组织化学研究

基本信息

批准号：
03454294
负责人：
IKEDA Kazuhiko
金额：
$ 2.88万
依托单位：
Tokyo Institute of Psychiatry
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1993
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03454294/
关键词：
schizophrenia brain dopamine dopamine receptor D_2 D_4 immunohistochemistry polymorphism

项目摘要

In order to investigate the possible involvement of dopamine receptor subtyeps in the pathogenesis of schizophrenia, we performed immunohistochemical studies of autopied brains by using anti-subtype specfic pepetide antibodies.We obtained rabbit antibodies to dopamine D2, D3, D4 receptors with high titers. Their specificity was verified by an ELISA assay. Anti-D2 and D4 antiboies immunostained various parts of rat brains, when the tisuues were fixed in Bouin solution and embedded in paraffin. We found the recovery of antigenicity of D4 receptor in formalin-fixed, parffinembedded brain sections after autoclaving the sections.We ezamined the hippocampal formation of brains of 9 mael patients with schizophrenia (according to DSM-III-R criteria) and 9 age-matched male persons. D4 immunoreactivity was mainly found around neurons of end plate and shafts of dendrorites of C3 pyramidal cells. That the immunoreactivity was localized in presynapsis was confirmed by a double immunostaining assay for D4 and synaptophisin. Despite of the close and intesive inspection we were unable to detect definit differences in the distribution, pattern or intensity of D4 immunoreactivity between tissues of schizophenics and controls.

为了研究多巴胺受体亚型在精神分裂症发病机制中的可能参与，我们使用抗亚型特异性肽抗体对尸检脑进行了免疫组织化学研究。我们获得了兔多巴胺D2、D3、D4受体的高效价抗体。它们的特异性通过酶联免疫吸附试验证实。将组织用Bouin溶液固定，石蜡包埋后，对大鼠脑各部位进行抗d2和D4抗体免疫染色。我们发现，在福尔马林固定、石蜡包埋的脑切片中，经过高压灭菌后，D4受体的抗原性恢复。我们检测了9名精神分裂症患者（符合DSM-III-R标准）和9名年龄匹配的男性的大脑海马结构。D4免疫反应主要发生在C3锥体细胞终板神经元和树突轴周围。D4和synaptophiin的双免疫染色证实了免疫反应性定位于突触前。尽管进行了密切和深入的检查，但我们无法发现精神分裂症患者和对照组组织之间D4免疫反应性的分布、模式或强度的明确差异。