Mallouk N, Jacquemond V, Allard B
Laboratoire de Physiologie des Eléments Excitables, Unité Mixte de Recherche, Centre National de la Recherche Scientifique 5578, Université Claude Bernard Lyon I, 43 Boulevard du 11 Novembre 1918, 69622 Villeurbanne Cedex, France.
Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4950-5. doi: 10.1073/pnas.97.9.4950.
Duchenne muscular dystrophy results from the lack of dystrophin, a cytoskeletal protein associated with the inner surface membrane, in skeletal muscle. The cellular mechanisms responsible for the progressive skeletal muscle degeneration that characterizes the disease are still debated. One hypothesis suggests that the resting sarcolemmal permeability for Ca(2+) is increased in dystrophic muscle, leading to Ca(2+) accumulation in the cytosol and eventually to protein degradation. However, more recently, this hypothesis was challenged seriously by several groups that did not find any significant increase in the global intracellular Ca(2+) in muscle from mdx mice, an animal model of the human disease. In the present study, using plasma membrane Ca(2+)-activated K(+) channels as subsarcolemmal Ca(2+) probe, we tested the possibility of a Ca(2+) accumulation at the restricted subsarcolemmal level in mdx skeletal muscle fibers. Using the cell-attached configuration of the patch-clamp technique, we demonstrated that the voltage threshold for activation of high conductance Ca(2+)-activated K(+) channels is significantly lower in mdx than in control muscle, suggesting a higher subsarcolemmal [Ca(2+)]. In inside-out patches, we showed that this shift in the voltage threshold for high conductance Ca(2+)-activated K(+) channel activation could correspond to a approximately 3-fold increase in the subsarcolemmal Ca(2+) concentration in mdx muscle. These data favor the hypothesis according to which an increased calcium entry is associated with the absence of dystrophin in mdx skeletal muscle, leading to Ca(2+) overload at the subsarcolemmal level.
杜兴氏肌肉营养不良症是由于骨骼肌中缺乏肌营养不良蛋白(一种与内膜表面相关的细胞骨架蛋白)所致。导致该疾病特征性进行性骨骼肌退化的细胞机制仍存在争议。一种假说认为,营养不良肌肉中静息时肌膜对Ca(2+)的通透性增加,导致Ca(2+)在细胞质中积累,最终导致蛋白质降解。然而,最近,这一假说受到了几个研究小组的严重质疑,他们在人类疾病的动物模型mdx小鼠的肌肉中未发现整体细胞内Ca(2+)有任何显著增加。在本研究中,我们使用质膜Ca(2+)激活的K(+)通道作为肌膜下Ca(2+)探针,测试了mdx骨骼肌纤维中肌膜下有限水平Ca(2+)积累的可能性。使用膜片钳技术的细胞贴附模式,我们证明mdx中高电导Ca(2+)激活的K(+)通道激活的电压阈值显著低于对照肌肉,表明肌膜下[Ca(2+)]更高。在内外膜片模式中,我们表明高电导Ca(2+)激活的K(+)通道激活电压阈值的这种变化可能对应于mdx肌肉中肌膜下Ca(2+)浓度增加约3倍。这些数据支持这样一种假说,即钙内流增加与mdx骨骼肌中缺乏肌营养不良蛋白有关,导致肌膜下水平Ca(2+)过载。