Yin Xiangling, Manczak Maria, Reddy P Hemachandra
Garrison Institute on Aging.
Garrison Institute on Aging, Cell Biology and Biochemistry, Neuroscience & Pharmacology, Neurology and Speech, Language and Hearing Sciences Departments, Texas Tech University Health Sciences Center, 3601 4th Street, MS 9424, Lubbock, TX 79430, USA
Hum Mol Genet. 2016 May 1;25(9):1739-53. doi: 10.1093/hmg/ddw045. Epub 2016 Feb 16.
The objective of this study was to determine the protective effects of the mitochondria-targeted molecules MitoQ and SS31 in striatal neurons that stably express mutant huntingtin (Htt) (STHDhQ111/Q111) in Huntington's disease (HD). We studied mitochondrial and synaptic activities by measuring mRNA and the protein levels of mitochondrial and synaptic genes, mitochondrial function, and ultra-structural changes in MitoQ- and SS31-treated mutant Htt neurons relative to untreated mutant Htt neurons. We used gene expression analysis, biochemical methods, transmission electron microscopy (TEM) and confocal microscopy methods. In the MitoQ- and SS31-treated mutant Htt neurons, fission genes Drp1 and Fis1 were down-regulated, and fusion genes Mfn1, Mfn2 and Opa1 were up-regulated relative to untreated neurons, suggesting that mitochondria-targeted molecules reduce fission activity. Interestingly, the mitochondrial biogenesis genes PGC1α, PGC1β, Nrf1, Nrf2 and TFAM were up-regulated in MitoQ- and SS31-treated mutant Htt neurons. The synaptic genes synaptophysin and PSD95 were up-regulated, and mitochondrial function was normal in the MitoQ- and SS31-treated mutant Htt neurons. Immunoblotting findings of mitochondrial and synaptic proteins agreed with the mRNA findings. TEM studies revealed decreased numbers of structurally intact mitochondria in MitoQ- and SS31-treated mutant Htt neurons. These findings suggest that mitochondria-targeted molecules MitoQ and SS31 are protective against mutant Htt-induced mitochondrial and synaptic damage in HD neurons, and these mitochondria-targeted molecules are potential therapeutic molecules for the treatment of HD neurons.
本研究的目的是确定线粒体靶向分子MitoQ和SS31对亨廷顿舞蹈病(HD)中稳定表达突变型亨廷顿蛋白(Htt)(STHDhQ111/Q111)的纹状体神经元的保护作用。我们通过测量线粒体和突触基因的mRNA和蛋白质水平、线粒体功能以及与未处理的突变型Htt神经元相比,MitoQ和SS31处理的突变型Htt神经元的超微结构变化,来研究线粒体和突触活动。我们使用了基因表达分析、生化方法、透射电子显微镜(TEM)和共聚焦显微镜方法。在MitoQ和SS31处理的突变型Htt神经元中,与未处理的神经元相比,裂变基因Drp1和Fis1下调,而融合基因Mfn1、Mfn2和Opa1上调,这表明线粒体靶向分子降低了裂变活性。有趣的是,在MitoQ和SS31处理的突变型Htt神经元中,线粒体生物发生基因PGC1α、PGC1β、Nrf1、Nrf2和TFAM上调。突触基因突触素和PSD95上调,并且在MitoQ和SS31处理的突变型Htt神经元中线粒体功能正常。线粒体和突触蛋白的免疫印迹结果与mRNA结果一致。TEM研究显示,在MitoQ和SS31处理的突变型Htt神经元中,结构完整的线粒体数量减少。这些发现表明,线粒体靶向分子MitoQ和SS31对HD神经元中突变型Htt诱导的线粒体和突触损伤具有保护作用,并且这些线粒体靶向分子是治疗HD神经元的潜在治疗分子。