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首页> 外文期刊>Neuromuscular disorders: NMD >Adenovirus-mediated expression of myogenic differentiation factor 1 (MyoD) in equine and human dermal fibroblasts enables their conversion to caffeine-sensitive myotubes
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Adenovirus-mediated expression of myogenic differentiation factor 1 (MyoD) in equine and human dermal fibroblasts enables their conversion to caffeine-sensitive myotubes

机译:腺病毒介导的马和人真皮成纤维细胞中肌源性分化因子1(MyoD)的表达使其能够转化为对咖啡因敏感的肌管

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摘要

Several human and animal myopathies, such as malignant hyperthermia (MH), central core disease and equine recurrent exertional rhabdomyolysis (RER) are confirmed or thought to be associated with dysfunction of skeletal muscle calcium regulation. For some patients in whom the genetic cause is unknown, or when mutational analysis reveals genetic variants with unclear pathogenicity, defects are further studied through use of muscle histopathology and in vitro contraction tests, the latter in particular, when assessing responses to ryanodine receptor agonists, such as caffeine. However, since muscle biopsy is not always suitable, researchers have used cultured cells to model these diseases, by examining calcium regulation in myotubes derived from skin, following forced expression of muscle-specific transcription factors. Here we describe a novel adenoviral vector that we used to express equine MyoD in dermal fibroblasts. In permissive conditions, transduced equine and human fibroblasts differentiated into multinucleated myotubes. We demonstrate that these cells have a functional excitation-calcium release mechanism and, similarly to primary muscle-derived myotubes, respond in a dose-dependent manner to increasing concentrations of caffeine. MyoD-induced conversion of equine skin-derived fibroblasts offers an attractive method for evaluating calcium homeostasis defects in vitro without the need for invasive muscle biopsy.
机译:几种人类和动物的肌病,例如恶性高热(MH),中枢核心疾病和马反复劳累性横纹肌溶解症(RER),已被确认或认为与骨骼肌钙调节功能障碍有关。对于某些遗传原因未知的患者,或者当突变分析显示出致病性不清楚的遗传变异时,将通过使用肌肉组织病理学和体外收缩试验来进一步研究缺陷,特别是在评估对莱诺丹定受体激动剂的反应时,例如咖啡因。但是,由于肌肉活检并不总是合适的,研究人员已使用培养的细胞来模拟这些疾病,方法是在强制表达肌肉特异性转录因子后,检查源自皮肤的肌管中的钙调节。在这里,我们描述了一种新颖的腺病毒载体,用于表达真皮成纤维细胞中的马MyoD。在允许的条件下,转导的马和人成纤维细胞分化为多核肌管。我们证明这些细胞具有功能性的兴奋性钙释放机制,并且类似于原发性肌肉衍生的肌管,以剂量依赖的方式对咖啡因浓度的增加作出反应。 MyoD诱导的马皮肤成纤维细胞转化提供了一种有吸引力的方法,可用于体外评估钙稳态平衡缺损,而无需进行侵入性肌肉活检。

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