Braubach Peter, Orynbayev Murat, Andronache Zoita, Hering Tanja, Landwehrmeyer Georg Bernhard, Lindenberg Katrin S, Melzer Werner
Institute of Applied Physiology and Department of Neurology, Ulm University, D-89081 Ulm, Germany.
Institute of Applied Physiology and Department of Neurology, Ulm University, D-89081 Ulm, Germany Institute of Applied Physiology and Department of Neurology, Ulm University, D-89081 Ulm, Germany.
J Gen Physiol. 2014 Nov;144(5):393-413. doi: 10.1085/jgp.201411255.
Huntington's disease (HD) is caused by an expanded CAG trinucleotide repeat within the gene encoding the protein huntingtin. The resulting elongated glutamine (poly-Q) sequence of mutant huntingtin (mhtt) affects both central neurons and skeletal muscle. Recent reports suggest that ryanodine receptor-based Ca(2+) signaling, which is crucial for skeletal muscle excitation-contraction coupling (ECC), is changed by mhtt in HD neurons. Consequently, we searched for alterations of ECC in muscle fibers of the R6/2 mouse, a mouse model of HD. We performed fluorometric recordings of action potentials (APs) and cellular Ca(2+) transients on intact isolated toe muscle fibers (musculi interossei), and measured L-type Ca(2+) inward currents on internally dialyzed fibers under voltage-clamp conditions. Both APs and AP-triggered Ca(2+) transients showed slower kinetics in R6/2 fibers than in fibers from wild-type mice. Ca(2+) removal from the myoplasm and Ca(2+) release flux from the sarcoplasmic reticulum were characterized using a Ca(2+) binding and transport model, which indicated a significant reduction in slow Ca(2+) removal activity and Ca(2+) release flux both after APs and under voltage-clamp conditions. In addition, the voltage-clamp experiments showed a highly significant decrease in L-type Ca(2+) channel conductance. These results indicate profound changes of Ca(2+) turnover in skeletal muscle of R6/2 mice and suggest that these changes may be associated with muscle pathology in HD.
亨廷顿舞蹈症(HD)由编码亨廷顿蛋白的基因内CAG三核苷酸重复序列扩增所致。突变型亨廷顿蛋白(mhtt)产生的延长型谷氨酰胺(多聚谷氨酰胺)序列会影响中枢神经元和骨骼肌。最近的报告表明,基于雷诺丁受体的Ca(2+)信号传导对骨骼肌兴奋 - 收缩偶联(ECC)至关重要,而在HD神经元中会被mhtt改变。因此,我们在HD小鼠模型R6/2的肌纤维中寻找ECC的改变。我们对完整分离的趾肌纤维(骨间肌)进行动作电位(APs)和细胞Ca(2+)瞬变的荧光记录,并在电压钳制条件下测量内部透析纤维上的L型Ca(2+)内向电流。与野生型小鼠的纤维相比,R6/2纤维中的APs和AP触发的Ca(2+)瞬变均显示出较慢的动力学。使用Ca(2+)结合和转运模型对肌浆中Ca(2+)的清除和肌浆网中Ca(2+)的释放通量进行了表征,结果表明,在APs后以及电压钳制条件下,缓慢的Ca(2+)清除活性和Ca(2+)释放通量均显著降低。此外,电压钳实验显示L型Ca(2+)通道电导显著降低。这些结果表明R6/2小鼠骨骼肌中Ca(2+)周转发生了深刻变化,并表明这些变化可能与HD中的肌肉病理有关。