JNK3-mediated apoptotic cell death in primary dopaminergic neurons.
JNK3-mediated apoptotic cell death in primary dopaminergic neurons.
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DOI：
10.1007/978-1-61779-170-3_19
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Xia, Zhengui
中科院分区：
文献类型：
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作者：
Choi, Won-Seok;Klintworth, Heather M;Xia, Zhengui
研究多巴胺能神经元死亡的机制对于了解帕金森病的发病机制至关重要，但这在技术上往往是相当具有挑战性的。在这里，我们描述了详细的方法，培养原代中脑多巴胺能神经元和检查c-Jun N-末端蛋白激酶（JNK）在这些文化的激活。我们利用免疫细胞化学和计算机分析来量化多巴胺能神经元存活的数量和多巴胺能神经元中JNK的激活。使用TUNEL染色来量化凋亡细胞死亡。siRNA用于特异性抑制JNK的神经特异性同种型JNK 3。我们的数据暗示JNK 3的激活在鱼藤酮诱导的多巴胺能神经元凋亡。
Investigation of mechanisms responsible for dopaminergic neuron death is critical for understanding the pathogenesis of Parkinson’s disease, yet this is often quite challenging technically. Here, we describe detailed methods for culturing primary mesencephalic dopaminergic neurons and examining the activation of c-Jun N-terminal protein Kinase (JNK) in these cultures. We utilized immunocytochemistry and computerized analysis to quantify the number of surviving dopaminergic neurons and JNK activation in dopaminergic neurons. TUNEL staining was used to quantify apoptotic cell death. siRNA was used to specifically inhibit JNK3, the neural specific isoform of JNK. Our data implicate the activation of JNK3 in rotenone-induced dopaminergic neuron apoptosis.