Devall Matthew, Burrage Joe, Caswell Richard, Johnson Matthew, Troakes Claire, Al-Sarraj Safa, Jeffries Aaron R, Mill Jonathan, Lunnon Katie
University of Exeter Medical School, University of Exeter, Devon, UK.
Institute of Psychiatry, King's College London, London, UK.
Biotechniques. 2015 Oct 1;59(4):241-2, 244-6. doi: 10.2144/000114343. eCollection 2015 Oct.
Given that many brain disorders are characterized by mitochondrial dysfunction, there is a growing interest in investigating genetic and epigenetic variation in mitochondrial DNA (mtDNA). One major caveat for such studies is the presence of nuclear-mitochondrial pseudogenes (NUMTs), which are regions of the mitochondrial genome that have been inserted into the nuclear genome over evolution and, if not accounted for, can confound genetic studies of mtDNA. Here we provide the first systematic comparison of methods for isolating mtDNA from frozen post-mortem human brain tissue. Our data show that a commercial method from Miltenyi Biotec, which magnetically isolates mitochondria using antibodies raised against the mitochondrial import receptor subunit TOM22, gives significant mtDNA enrichment and should be considered the method of choice for mtDNA studies in frozen brain tissue.
鉴于许多脑部疾病的特征是线粒体功能障碍，人们对研究线粒体DNA（mtDNA）的遗传和表观遗传变异的兴趣日益浓厚。此类研究的一个主要问题是存在核线粒体假基因（NUMTs），这些假基因是线粒体基因组中在进化过程中插入到核基因组中的区域，如果不加以考虑，可能会混淆mtDNA的遗传研究。在此，我们首次对从冷冻的死后人类脑组织中分离mtDNA的方法进行了系统比较。我们的数据表明，Miltenyi Biotec公司的一种商业方法，即使用针对线粒体输入受体亚基TOM22产生的抗体通过磁性分离线粒体，能显著富集mtDNA，应被视为冷冻脑组织中mtDNA研究的首选方法。
