Roy Roland R, Pierotti David J, Garfinkel Alan, Zhong Hui, Baldwin Kenneth M, Edgerton V Reggie
Brain Research Institute, University of California, Los Angeles, CA 90024-1761, USA.
J Exp Biol. 2008 Apr;211(Pt 7):1041-9. doi: 10.1242/jeb.013722.
The clarity of categorizing skeletal muscle fibers in individual motor units into phenotypes based on quantitative single fiber enzyme activities and as a function of neuromuscular activity level was examined. Neuromuscular activity was eliminated in adult cat hindlimb muscles by spinal cord isolation (SI), i.e. complete spinal cord transection at a low thoracic and a high sacral level with bilateral dorsal rhizotomy between the transection sites. One motor unit was isolated via ventral root teasing procedures from the tibialis anterior (TA) muscle of each hindlimb in control and SI cats, and physiologically tested and glycogen depleted through repetitive stimulation; fibers comprising each motor unit were visualized through glycogen staining. Each motor unit was composed of fibers of the same myosin immunohistochemical type. Myofibrillar adenosine triphosphatase, succinate dehydrogenase and alpha-glycerophosphate dehydrogenase activities were determined for a sample of motor unit and non-motor unit fibers, providing a measure of three enzyme activities often used to characterize fiber phenotype within a single unit. Although normal enzyme activities were altered after 6 months of inactivity, the relationships among the three enzymes were largely maintained. These data demonstrate that it is not the diversity in any single enzyme property but the profile of several metabolic pathways that underlies the significance of fiber phenotypes. These profiles must reflect a high level of coordination of expression of selected combinations of genes. Although neuromuscular activity level influences fiber phenotype, the present results demonstrate that activity-independent mechanisms remain important sources of the control of phenotype establishment in the near absence of activity.
基于定量单纤维酶活性并作为神经肌肉活动水平的函数,对单个运动单位中的骨骼肌纤维进行表型分类的清晰度进行了研究。通过脊髓隔离(SI)消除成年猫后肢肌肉中的神经肌肉活动,即在低胸段和高骶段进行完全脊髓横断,并在横断部位之间进行双侧背根切断术。通过腹根剥离程序从对照猫和脊髓隔离猫的每个后肢的胫骨前肌(TA)中分离出一个运动单位,进行生理测试并通过重复刺激使糖原耗尽;通过糖原染色观察每个运动单位的纤维。每个运动单位由相同肌球蛋白免疫组织化学类型的纤维组成。测定运动单位纤维和非运动单位纤维样本的肌原纤维三磷酸腺苷酶、琥珀酸脱氢酶和α-甘油磷酸脱氢酶活性,提供通常用于表征单个单位内纤维表型的三种酶活性的测量值。尽管6个月不活动后正常酶活性发生了改变,但三种酶之间的关系在很大程度上得以维持。这些数据表明,构成纤维表型重要性基础的不是任何单一酶特性的多样性,而是几种代谢途径的概况。这些概况必须反映所选基因组合表达的高度协调性。尽管神经肌肉活动水平会影响纤维表型,但目前的结果表明,在几乎没有活动的情况下,与活动无关的机制仍然是控制表型建立的重要来源。