Buck Eva, Zügel Martina, Schumann Uwe, Merz Tamara, Gumpp Anja M, Witting Anke, Steinacker Jürgen M, Landwehrmeyer G Bernhard, Weydt Patrick, Calzia Enrico, Lindenberg Katrin S
Department of Neurology, Ulm University, Ulm, Germany.
Division of Sports- and Rehabilitation Medicine, Ulm University Medical Center, Ulm, Germany.
PLoS One. 2017 Apr 13;12(4):e0175248. doi: 10.1371/journal.pone.0175248. eCollection 2017.
Alterations in mitochondrial respiration are an important hallmark of Huntington's disease (HD), one of the most common monogenetic causes of neurodegeneration. The ubiquitous expression of the disease causing mutant huntingtin gene raises the prospect that mitochondrial respiratory deficits can be detected in skeletal muscle. While this tissue is readily accessible in humans, transgenic animal models offer the opportunity to cross-validate findings and allow for comparisons across organs, including the brain. The integrated respiratory chain function of the human vastus lateralis muscle was measured by high-resolution respirometry (HRR) in freshly taken fine-needle biopsies from seven pre-manifest HD expansion mutation carriers and nine controls. The respiratory parameters were unaffected. For comparison skeletal muscle isolated from HD knock-in mice (HdhQ111) as well as a broader spectrum of tissues including cortex, liver and heart muscle were examined by HRR. Significant changes of mitochondrial respiration in the HdhQ knock-in mouse model were restricted to the liver and the cortex. Mitochondrial mass as quantified by mitochondrial DNA copy number and citrate synthase activity was stable in murine HD-model tissue compared to control. mRNA levels of key enzymes were determined to characterize mitochondrial metabolic pathways in HdhQ mice. We demonstrated the feasibility to perform high-resolution respirometry measurements from small human HD muscle biopsies. Furthermore, we conclude that alterations in respiratory parameters of pre-manifest human muscle biopsies are rather limited and mirrored by a similar absence of marked alterations in HdhQ skeletal muscle. In contrast, the HdhQ111 murine cortex and liver did show respiratory alterations highlighting the tissue specific nature of mutant huntingtin effects on respiration.
线粒体呼吸的改变是亨廷顿舞蹈症(HD)的一个重要标志,HD是神经退行性变最常见的单基因病因之一。致病突变亨廷顿基因的广泛表达使人想到,在骨骼肌中可能检测到线粒体呼吸功能缺陷。虽然在人类中这种组织很容易获取,但转基因动物模型提供了交叉验证研究结果的机会,并允许对包括大脑在内的不同器官进行比较。通过高分辨率呼吸测定法(HRR)测量了7名HD前临床期扩增突变携带者和9名对照者新鲜采集的股外侧肌细针活检样本中人类股外侧肌的呼吸链综合功能。呼吸参数未受影响。为作比较,通过HRR检查了从HD基因敲入小鼠(HdhQ111)分离出的骨骼肌以及更广泛的组织,包括皮质、肝脏和心肌。在HdhQ基因敲入小鼠模型中,线粒体呼吸的显著变化仅限于肝脏和皮质。与对照相比,通过线粒体DNA拷贝数和柠檬酸合酶活性定量的线粒体质量在小鼠HD模型组织中是稳定的。测定了关键酶的mRNA水平,以表征HdhQ小鼠的线粒体代谢途径。我们证明了从小的人类HD肌肉活检样本中进行高分辨率呼吸测定的可行性。此外,我们得出结论,HD前临床期人类肌肉活检样本呼吸参数的改变相当有限,并且在HdhQ骨骼肌中也同样没有明显改变。相反,HdhQ111小鼠的皮质和肝脏确实出现了呼吸改变,突出了突变亨廷顿蛋白对呼吸作用影响的组织特异性。