6-OHDA损毁的帕金森病大鼠模型纹状体中差异表达基因的筛选和功能研究

发布时间：2018-03-23 18:33

本文选题：帕金森病　切入点：cystatin　出处：《中国科学院研究生院（上海生命科学研究院）》2005年博士论文

【摘要】：我们通过microarray 和消减抑制杂交的方法获得帕金森病纹状体内差异表达的200 左右基因,经Northern、RT-PCR、原位杂交和免疫组化进一步鉴定,获得上调及下调的基因18 个。我们对其中几个感兴趣的基因cystatin C,trophinin,bystin 做了进一步的研究。 Cystatin C 是II 型半胱氨酸蛋白酶(cysteine protease)抑制剂家族中(cystatins)的一个成员, Northern blot 结果显示它在6-OHDA 损毁的大鼠纹状体中持续上调。原位杂交和免疫组化结果也同样证实Cystatin C 在6-OHDA 损毁的大鼠纹状体中差异表达,免疫荧光双标结果表明它来源于多种细胞成分,即神经元、星型胶质细胞和小胶质细胞。Cystatin C 对体外培养的E14 腹侧中脑多巴胺能神经元有促进存活作用。体内实验同样证实了cystatin C 对6-OHDA 损毁的大鼠多巴胺能神经元的存活作用。 Trophinin 和Bystin 曾被报道作为一个在胚胎着床过程中起重要作用的粘附分子,但该分子在中枢神经系统中的存在和作用从未有过报道。我们的结果表明在6-OHDA 损伤的PD 大鼠中,Trophinin 和bystin 在黑质和纹状体中的表达具有明显的时空特点,提示它们在中枢神经系统具有某种作用。
[Abstract]:We obtained about 200 differentially expressed genes in the striatum of Parkinson's disease by microarray and subtractive suppression hybridization, which were further identified by Northern RT-PCR, in situ hybridization and immunohistochemistry. A total of 18 up-regulated and down-regulated genes were obtained, and several of them were further studied. Cystatin C is a member of the cysteine protease II inhibitor family. Northern blot results show that Cystatin C is up-regulated in the striatum damaged by 6-OHDA. The results of in situ hybridization and immunohistochemistry also confirm the presence of Cystatin C in 6-OHDA. Differential expression in the damaged rat striatum, Immunofluorescence double labeling results showed that it originated from a variety of cell components, namely neurons, Astrocytes and microglia. Cystatin C promoted the survival of dopaminergic neurons in the ventral mesencephalon of E14 cultured in vitro. The in vivo experiments also confirmed the survival of cystatin C on the dopaminergic neurons damaged by 6-OHDA in rats. Trophinin and Bystin have been reported as a adhesion molecule that plays an important role in embryo implantation. However, the presence and role of this molecule in the central nervous system have never been reported. Our results show that the expression of Trophinin and bystin in substantia nigra and striatum of PD rats damaged by 6-OHDA has obvious temporal and spatial characteristics. These results suggest that they play a role in the central nervous system.
【学位授予单位】：中国科学院研究生院（上海生命科学研究院）
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R-332

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