MPTP诱导的褐鼠黑质基因差异表达的研究

本研究目的在于获取１－甲基－４－苯基－１,２,３,６－四氢吡啶（ＭＰＴＰ）诱导的褐鼠黑质差异表达的表达序列标签（ＥＳＴ）。通过ＭＰＴＰ腹腔注射建立褐鼠帕金森病模型,利用差别显示反转录ＰＣＲ（ＤＤＲＴ－ ＰＣＲ）技术分析ＭＰＴＰ 处理褐鼠与正常褐鼠黑质基因差异表达的状况,并筛选差异表达基因片段。结果经ＤＤＲＴ－ＰＣＲ 筛选并通过反Ｎｏｒｔｈｅｒｎ 杂交去除假阳性,最终获得２ 个差异表达基因片段。对其中一个片段进行克隆、测序和同源性比较发现,该片段为一个新的ＥＳＴ 序列,并且与胰岛素原样α肽具有同源性。本研究的结果提示,差异表达序列的分析有可能为研究ＭＰＴＰ诱导的褐鼠黑质变性的分子机理提供线索。

MPTP-INDUCED DIFFERENTIALLY EXPRESSED GENE IN SUBSTANTIA NIGRA OF C57BL MICE

The purpose of the present work is to analyze the 1 methyl 4 phenyl 1,2,3,6 tetrahydropyridine(MPTP) induced differential gene expression in the substantia nigra of C57BL mice and to screen Parkinson's disease related expressed sequence tag(EST). Parkinson's disease model of C57BL mice was established by consecutive administration of MPTP. Differential display reverse transcription polymerase chain reaction(DDRT PCR) was carried out to screen the differentially expressed gene fragment between the substantia nigra of the experimental and control groups. Two differentially expressed fragments were obtained and proved to be true positive with reverse Northern assay. One of these two fragments was found to be a new EST and showed 46% amino acid sequence homology with pro insulin like alpha peptide. There was a clue that the pro insulin like alpha peptide homologous EST may play certain roles in the MPTP induced substantia nigra degeneration, but it must be confirmed by further experiments.