TRPC1-mediated inhibition of 1-methyl-4-phenylpyridinium ion neurotoxicity in human SH-SY5Y neuroblastoma cells
TRPC1-mediated inhibition of 1-methyl-4-phenylpyridinium ion neurotoxicity in human SH-SY5Y neuroblastoma cells
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DOI：
10.1074/jbc.m407384200
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发表时间：
2005-01-21
影响因子：
4.8
通讯作者：
Ebadi, M
中科院分区：
文献类型：
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作者：
Bollimuntha, S;Singh, BB;Ebadi, M
果蝇典型瞬时受体电位（TRP）蛋白的哺乳动物同源物被认为具有质膜Ca 2+通道的功能。本研究检测了TRPC 1在人神经母细胞瘤（SH-SY 5 Y）细胞中的作用。用外源性神经毒素1-甲基-4-苯基吡啶离子（MPP+）处理的SH-SY 5 Y细胞显著降低TRPC 1蛋白水平。用MPP+处理的SH-SY 5 Y细胞的共聚焦显微镜显示TRPC 1的质膜染色减少。重要的是，TRPC 1的过表达减少了MPP+诱导的神经毒性。MPP+诱导的α-突触核蛋白表达也被TRPC 1过表达抑制。过表达反义TRPC 1 cDNA构建体或加入非特异性瞬时受体电位通道阻断剂镧后，SH-SY 5 Y细胞对MPP+的保护作用显著降低。TRPC 1的激活thapsigargin或卡巴胆碱降低MPP+神经毒性，这是部分依赖于外部Ca 2+。用凋亡标记物（Y 0-PRO-1）染色SH-SY 5 Y细胞显示TRPC 1保护SH-SY 5 Y神经元细胞免于凋亡。此外，TRPC 1过表达抑制细胞色素c的释放，并降低Bax和Apaf-1蛋白水平。对上述数据的解释表明，MPP+处理后TRPC 1细胞表面表达的减少可能与多巴胺能神经变性有关。此外，TRPC 1可以抑制退行性凋亡信号传导，以提供针对帕金森病诱导剂的神经保护。
Mammalian homologues of the Drosophila canonical transient receptor potential (TRP) proteins have been implicated to function as plasma membrane Ca2+ channels. This study examined the role of TRPC1 in human neuroblastoma (SH- SY5Y)cells. SH- SY5Y cells treated with an exogenous neurotoxin, 1-methyl-4-phenylpyridinium ion (MPP+) significantly decreased TRPC1 protein levels. Confocal microscopy on SH-SY5Y cells treatment with MPP+ showed decreased plasma membrane staining of TRPC1. Importantly, overexpression of TRPC1 reduced neurotoxicity induced by MPP+. MPP+-induced alpha-synuclein expression was also suppressed by TRPC1 overexpression. Protection of SH- SY5Y cells against MPP+ was significantly decreased upon the overexpression of antisense TRPC1 cDNA construct or the addition of a nonspecific transient receptor potential channel blocker lanthanum. Activation of TRPC1 by thapsigargin or carbachol decreased MPP+ neurotoxicity, which was partially dependent on external Ca2+. Staining of SH-SY5Y cells with an apoptotic marker (YO-PRO-1) showed that TRPC1 protects SH- SY5Y neuronal cells against apoptosis. Further, TRPC1 overexpression inhibited cytochrome c release and decreased Bax and Apaf-1 protein levels. Interpretation of the above data suggests that reduction in the cell surface expression of TRPC1 following MPP+ treatment may be involved in dopaminergic neurodegeneration. Furthermore, TRPC1 may inhibit degenerative apoptotic signaling to provide neuroprotection against Parkinson's disease-inducing agents.