Tyrosine hydroxylase and dopamine transporter expression following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurodegeneration of the mouse nigrostriatal pathway
Tyrosine hydroxylase and dopamine transporter expression following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurodegeneration of the mouse nigrostriatal pathway
复制标题
DOI：
10.1002/jnr.20114
复制
发表时间：
2004-05-15
影响因子：
4.2
通讯作者：
Petzinger, GM
中科院分区：
文献类型：
--
作者：
Jakowec, MW;Nixon, K;Petzinger, GM
给予C57 BL/6小鼠神经毒剂1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP）靶向黑质纹状体多巴胺能神经元，导致细胞死亡和纹状体多巴胺耗竭。在年轻成年小鼠MPTP损伤后，存活的黑质纹状体多巴胺能神经元在损伤后数周至数月显示出纹状体多巴胺的稳健且可重复的恢复。因此，小鼠提供了一个很好的模型，以探讨神经毒性损伤后黑质纹状体系统的神经可塑性的机制。本研究的目的是分析蛋白质和mRNA转录的基因参与多巴胺的生物合成（酪氨酸羟化酶; TH）和摄取（多巴胺转运蛋白; DAT）方面的时间进程（7-90天）MPTP损伤后。使用免疫组织化学和Western免疫印迹技术的分子分析表明，纹状体TH增加30-60天postlesioning，返回到接近控制（prelesioned）水平的60-90天。原位杂交组织化学表明，TH蛋白的增加可能是由于部分增加TH mRNA的表达在幸存的黑质纹状体多巴胺能神经元。TH蛋白在7，30，60，和90天postlesioning与二维聚丙烯酰胺凝胶电泳结合Western免疫印迹分析显示改变TH蛋白亚型迁移在等电点不同于那些天然亚型。相反，TH蛋白，恢复到pre-lesioned水平，60天，DAT蛋白分析表明，纹状体DAT蛋白的表达增加，直到90天postlesioning没有恢复到接近pre-lesion水平。这些结果表明，TH和DAT可能在存活的多巴胺能神经元的表达的时间过程中不同，并可能在介导纹状体多巴胺的返回中发挥作用。(C)2004 Wiley-Liss，Inc.
Administration of the neurotoxicant 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to C57BL/6 mice targets nigrostriatal dopaminergic neurons, leading to cell death and the depletion of striatal dopamine. After MPTP lesioning in young adult mice, surviving nigrostriatal dopaminergic neurons display robust and reproducible return of striatal dopamine weeks to months after injury. Thus, the mouse provides an excellent model with which to investigate the mechanisms underlying neuroplasticity of the nigrostriatal system following neurotoxic injury. The purpose of this study was to analyze proteins and mRNA transcripts of genes involved in dopamine biosynthesis (tyrosine hydroxylase; TH) and uptake (dopamine transporter; DAT) with regard to time course (7-90 days) after MPTP lesioning. Molecular analysis using immunohistochemistry and Western immunoblotting techniques demonstrated an increase in striatal TH by 30-60 days postlesioning that returned to near-control (prelesioned) levels by 60-90 days. In situ hybridization histochemistry indicated that this increase in TH protein might be due in part to increased TH mRNA expression in surviving nigrostriatal dopaminergic neurons. Analysis of TH protein at 7, 30, 60, and 90 days postlesioning with two-dimensional polyacrylamide gel electrophoresis in conjunction with Western immunoblotting revealed altered TH protein isoforms migrating at isoelectric points different from those of the native isoform. In contrast to TH protein, which returned to prelesioned levels by 60 days, DAT protein analysis showed that increased expression of striatal DAT protein did not return to near-prelesion levels until 90 days postlesioning. These results suggest that TH and DAT may differ in their time course of expression in surviving dopaminergic neurons and may play a role in mediating the return of striatal dopamine. (C) 2004 Wiley-Liss, Inc.