6-Hydroxydopamine (6-OHDA) induces Drp1-dependent mitochondrial fragmentation in SH-SY5Y cells
6-Hydroxydopamine (6-OHDA) induces Drp1-dependent mitochondrial fragmentation in SH-SY5Y cells
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DOI：
10.1016/j.freeradbiomed.2008.03.009
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发表时间：
2008-06-01
影响因子：
7.4
通讯作者：
Schrader, Michael
中科院分区：
文献类型：
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作者：
Gomez-Lazaro, Maria;Bonekamp, Nina A.;Schrader, Michael
线粒体改变与6-羟基多巴胺（6-OHDA）的细胞毒性作用有关，6-羟基多巴胺是一种广泛用于帕金森病研究的神经毒素。我们研究了6-OHDA对SH-SY5Y神经母细胞瘤细胞线粒体形态的潜在影响。通过免疫荧光和延时荧光显微镜，我们发现6-OHDA在SH-SY5Y细胞中诱导了严重的线粒体断裂，这一事件类似于动力蛋白相关蛋白1 （DLP1/Drp1）的膜接体Fis1p的过表达诱导的线粒体分裂。6-OHDA未引起过氧化物酶体形态的任何变化。生化实验表明，6- ohda诱导的线粒体断裂是SH-SY5Y细胞线粒体膜电位崩溃和细胞色素c释放之前的早期事件。参与线粒体和过氧化物酶体裂变的DLP1/Drp1的沉默阻止了6- ohda诱导的线粒体断裂。此外，在Drp1沉默的细胞中，6-OHDA诱导的细胞死亡减少，这表明线粒体分裂的阻断保护SH-SY5Y细胞免受6-OHDA的毒性。在缺乏Bax或p53的小鼠胚胎成纤维细胞中进行的实验表明，这两种蛋白对于6-羟多巴胺诱导的线粒体断裂并不是必需的。我们的数据首次证明线粒体断裂和Drp1功能参与6- ohda诱导的细胞凋亡。Elsevier Inc.出版。
Mitochondrial alterations have been associated with the cytotoxic effect of 6-hydroxydopamine (6-OHDA), a widely used neurotoxin to study Parkinson's disease. Herein we studied the potential effects of 6-OHDA on mitochondrial morphology in SH-SY5Y neuroblastoma cells. By immunofluorescence and time-lapse fluorescence microscopy we demonstrated that 6-OHDA induced profound mitochondrial fragmentation in SH-SY5Y cells, an event that was similar to mitochondrial fission induced by overexpression of Fis1p, a membrane adaptor for the dynamin-related protein 1 (DLP1/Drp1). 6-OHDA failed to induce any changes in peroxisome morphology. Biochemical experiments revealed that 6-OHDA-induced mitochondrial fragmentation is an early event preceding the collapse of the mitochondrial membrane potential and cytochrome c release in SH-SY5Y cells. Silencing of DLP1/Drp1, which is involved in mitochondrial and peroxisomal fission, prevented 6-OHDA-induced fragmentation of mitochondria. Furthermore, in cells silenced for Drp1, 6-OHDA-induced cell death was reduced, indicating that a block in mitochondrial fission protects SH-SY5Y cells against 6-OHDA toxicity. Experiments in mouse embryonic fibroblasts deficient in Bax or p53 revealed that both proteins are not essential for 6-OHDA-induced mitochondrial fragmentation. Our data demonstrate for the first time an involvement of mitochondrial fragmentation and Drp1 function in 6-OHDA-induced apoptosis. Published by Elsevier Inc.