Vincent Amy E, Grady John P, Rocha Mariana C, Alston Charlotte L, Rygiel Karolina A, Barresi Rita, Taylor Robert W, Turnbull Doug M
Wellcome Trust Centre for Mitochondrial Research, Institute of Neuroscience, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
Rare Diseases Advisory Group Service for Neuromuscular Diseases, Muscle Immunoanalysis Unit, Newcastle upon Tyne Hospitals NHS Foundation Trust, Newcastle upon Tyne, NE2 4AZ, UK.
Neuromuscul Disord. 2016 Oct;26(10):691-701. doi: 10.1016/j.nmd.2016.08.004. Epub 2016 Aug 10.
Myofibrillar myopathies (MFM) are characterised by focal myofibrillar destruction and accumulation of myofibrillar elements as protein aggregates. They are caused by mutations in the DES, MYOT, CRYAB, FLNC, BAG3, DNAJB6 and ZASP genes as well as other as yet unidentified genes. Previous studies have reported changes in mitochondrial morphology and cellular positioning, as well as clonally-expanded, large-scale mitochondrial DNA (mtDNA) deletions and focal respiratory chain deficiency in muscle of MFM patients. Here we examine skeletal muscle from patients with desmin (n = 6), ZASP (n = 1) and myotilin (n = 2) mutations and MFM protein aggregates, to understand how mitochondrial dysfunction may contribute to the underlying mechanisms causing disease pathology. We have used a validated quantitative immunofluorescent assay to study respiratory chain protein levels, together with oxidative enzyme histochemistry and single cell mitochondrial DNA analysis, to examine mitochondrial changes. Results demonstrate a small number of clonally-expanded mitochondrial DNA deletions, which we conclude are due to both ageing and disease pathology. Further to this we report higher levels of respiratory chain complex I and IV deficiency compared to age matched controls, although overall levels of respiratory deficient muscle fibres in patient biopsies are low. More strikingly, a significantly higher percentage of myofibrillar myopathy patient muscle fibres have a low mitochondrial mass compared to controls. We concluded this is mechanistically unrelated to desmin and myotilin protein aggregates; however, correlation between mitochondrial mass and muscle fibre area is found. We suggest this may be due to reduced mitochondrial biogenesis in combination with muscle fibre hypertrophy.
肌原纤维肌病(MFM)的特征是局灶性肌原纤维破坏以及肌原纤维成分以蛋白质聚集体的形式积累。它们由DES、MYOT、CRYAB、FLNC、BAG3、DNAJB6和ZASP基因以及其他尚未确定的基因突变引起。先前的研究报道了MFM患者肌肉中线粒体形态和细胞定位的变化,以及克隆性扩增的大规模线粒体DNA(mtDNA)缺失和局灶性呼吸链缺陷。在这里,我们检查了患有结蛋白(n = 6)、ZASP(n = 1)和肌联蛋白(n = 2)突变以及MFM蛋白聚集体的患者的骨骼肌,以了解线粒体功能障碍如何可能导致疾病病理的潜在机制。我们使用了经过验证的定量免疫荧光测定法来研究呼吸链蛋白水平,同时结合氧化酶组织化学和单细胞线粒体DNA分析,以检查线粒体变化。结果显示存在少量克隆性扩增的线粒体DNA缺失,我们认为这是由于衰老和疾病病理共同导致的。除此之外,我们报告称,与年龄匹配的对照组相比,呼吸链复合体I和IV缺陷水平更高,尽管患者活检中呼吸缺陷肌纤维的总体水平较低。更引人注目的是,与对照组相比,肌原纤维肌病患者肌肉纤维中线粒体质量低的比例显著更高。我们得出结论,这在机制上与结蛋白和肌联蛋白蛋白聚集体无关;然而,发现线粒体质量与肌纤维面积之间存在相关性。我们认为这可能是由于线粒体生物发生减少与肌纤维肥大共同作用的结果。