Analysis of targeted mutation in DJ-1 on cellular function in primary astrocytes.
Analysis of targeted mutation in DJ-1 on cellular function in primary astrocytes.
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DOI：
10.1016/j.toxlet.2008.11.008
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发表时间：
2009-02-10
影响因子：
3.5
通讯作者：
Legare ME
中科院分区：
文献类型：
--
作者：
Ashley AK;Hanneman WH;Katoh T;Moreno JA;Pollack A;Tjalkens RB;Legare ME
DJ-1突变诱导早发性帕金森病，相反，DJ-1的过度表达与许多组织中的癌症相关。利用在胚胎干细胞中进行的基因捕获筛选文库来产生DJ-1突变小鼠。采用实时荧光定量PCR和免疫印迹技术检测DJ-1基因的功能突变。成年小鼠组织中的正常DJ-1蛋白表达被表征，并且在脑组织中表现出高表达，具有广泛的全身分布。从DJ-1−/−小鼠中分离的原代星形胶质细胞显示，响应于鱼藤酮或LPS，DJ-1蛋白的核定位减少，伴随着仅在鱼藤酮暴露中发现的DJ-1线粒体定位的增加。与对照组相比，DJ-1−/−星形胶质细胞的静息线粒体膜电位显著降低。我们的DJ-1基因敲除小鼠为探索DJ-1的分子和生理作用提供了一个令人兴奋的工具，以进一步阐明其在神经退行性变中的功能。
DJ-1 mutation induces early-onset Parkinson's disease, and conversely over-expression of DJ-1 is associated with cancer in numerous tissues. A gene-trap screening library conducted in embryonic stem cells was utilized for generation of a DJ-1 mutant mouse. Real-time PCR and immunoblotting were utilized to confirm functional mutation of the DJ-1 gene. Normal DJ-1 protein expression in adult mouse tissue was characterized and demonstrates high expression in brain tissue with wide systemic distribution. Primary astrocytes isolated from DJ-1−/− mice reveal a decreased nuclear localization of DJ-1 protein in response to rotenone or LPS, with a concomitant increase in mitochondrial localization of DJ-1 found only in the rotenone exposure. Resting mitochondrial membrane potential was significantly lower in DJ-1−/− astrocytes, as compared to controls. Our DJ-1 knockout mouse provides an exciting tool for exploring the molecular and physiological roles of DJ-1 to further explicate its functions in neurodegeneration.