A novel quantitative morphometry approach to assess regeneration in dystrophic skeletal muscle
A novel quantitative morphometry approach to assess regeneration in dystrophic skeletal muscle
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DOI：
10.1016/j.nmd.2014.04.011
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发表时间：
2014-07-01
影响因子：
2.8
通讯作者：
Friedrich, Oliver
中科院分区：
文献类型：
--
作者：
Buttgereit, Andreas;Weber, Cornelia;Friedrich, Oliver
杜氏肌营养不良症是一种遗传性退行性肌肉疾病，伴有骨骼肌和心肌的进行性无力。受干扰的钙稳态和信号通路导致肌肉组织纤维化重塑的变性/再生周期，并由慢性炎症维持。除了微结构改变外，营养不良肌纤维的再生通常仅根据位于中心的细胞核进行分类，但缺乏再生过程与核体积、肌球蛋白数量、结构和功能质量的相关性，特别是在再生能力耗尽的老肌肉中。这些信息可以产生新的再生功能生物标志物。在这里，我们使用二次谐波产生和多光子荧光显微镜对表达Delta ex 17-48微型肌营养不良蛋白的野生型、营养不良mdx和转基因mdx小鼠的完整单个肌纤维进行检测，以确定中心核纤维的百分比和核与肌球蛋白体积比与年龄的关系。基于这个比率，我们将“生物动力效率”定义为纤维成熟的光学测量，在成年野生型和迷你肌营养不良蛋白纤维中接近统一，但在非常年轻和年老的mdx小鼠中较小，因为正在进行的细胞成熟（年轻）和再生（老年）。有了这些参数，就有可能提供关于肌纤维再生的定量测量。(C) 2014 Elsevier B.V.版权所有
Duchenne muscular dystrophy is an inherited degenerative muscle disease with progressive weakness of skeletal and cardiac muscle. Disturbed calcium homeostasis and signalling pathways result in degeneration/regeneration cycles with fibrotic remodelling of muscle tissue, sustained by chronic inflammation. In addition to altered microarchitecture, regeneration in dystrophic muscle fibres is often only classified by centrally located nuclei but correlation of the regeneration process to nuclear volumes, myosin amounts, architecture and functional quality are missing, in particular in old muscles where the regenerative capacity is exhausted. Such information could yield novel regeneration-to-function biomarkers. Here we used second harmonic generation and multi photon fluorescence microscopy in intact single muscle fibres from wild-type, dystrophic mdx and transgenic mdx mice expressing an Delta ex 17-48 mini-dystrophin to determine the percentage of centronucleated fibres and nucleus-to-myosin volume ratio as a function of age. Based on this ratio we define a `biomotoric efficiency' as an optical measure for fibre maturation, which is close to unity in adult wild-type and mini-dystrophin fibres, but smaller in very young and old mdx mice as a result of ongoing cell maturation (young) and regeneration (aged). With these parameters it is possible to provide a quantitative measure about muscle fibre regeneration. (C) 2014 Elsevier B.V. All rights reserved.