Immunochemical analysis of dopamine transporter protein in Parkinson's disease
Immunochemical analysis of dopamine transporter protein in Parkinson's disease
复制标题
DOI：
10.1002/ana.410410417
复制
发表时间：
1997-04-01
影响因子：
11.2
通讯作者：
Levey, AI
中科院分区：
文献类型：
--
作者：
Miller, GW;Staley, JK;Levey, AI
质膜多巴胺转运蛋白（DAT）被认为是突触前多巴胺能末端丧失的可靠标志。先前的体内成像和死后结合研究已经检测到帕金森病（PD）大脑中纹状体DAT结合的缺失；然而，这些技术的空间分辨率较差，并且可能受到某些配体非特异性结合的影响。在这项研究中，我们使用新的高度特异性的单克隆抗体对人DAT的不同表位进行定量和定位。Western blot分析显示，PD脑壳核、尾状核和伏隔核的DAT免疫反应性与对照组相比明显降低，且与病程显著相关。免疫组织化学显示，在对照组的纹状体中，dat免疫反应纤维和点密集，但在PD脑的壳核中，这些纤维和点明显减少。PD脑尾状核沿脑室边缘明显保留，伏隔核相对保留。一个意想不到的发现是在PD壳核基质中保留了离散的DAT免疫反应岛。因此，DAT蛋白的免疫学分析为PD和其他涉及多巴胺能系统的神经系统疾病的DAT蛋白定位和定量提供了新的和敏感的手段。
The plasma membrane dopamine transporter (DAT) is considered to be a reliable marker of presynaptic dopaminergic terminal loss. Previous in vivo imaging and postmortem binding studies have detected a loss in striatal DAT binding in Parkinson's diseased (PD) brain; however, these techniques have poor spatial resolution and may suffer from nonspecific binding of some ligands. In this study, we use novel highly specific monoclonal antibodies to distinct epitopes of human DAT to quantify and localize the protein. Western blot analysis revealed marked reductions in DAT immunoreactivity in putamen, caudate, and nucleus accumbens of PD brain compared with control cases, and the reductions were significantly correlated to disease duration. Immunohistochemistry revealed DAT-immunoreactive fibers and puncta that were dense throughout the striatum of control brains but that were drastically reduced in putamen of PD brains. Caudate from PD brains showed a significant degree of sparing along the border of the ventricle, and the nucleus accumbens was relatively preserved. An unexpected finding was that discrete islands of DAT immunoreactivity were preserved within the matrix of PD putamen. Thus, immunological analysis of DAT protein provides novel and sensitive means for localizing and quantifying DAT protein in PD and other neurological disorders involving dopaminergic systems.