Ellefsen S, Vikmoen O, Zacharoff E, Rauk I, Slettaløkken G, Hammarström D, Strand T A, Whist J E, Hanestadhaugen M, Vegge G, Fagernes C E, Nygaard H, Hollan I, Rønnestad B R
Lillehammer University College, Lillehammer, Norway.
Scand J Med Sci Sports. 2014 Oct;24(5):e332-42. doi: 10.1111/sms.12185. Epub 2014 Feb 20.
Determination of muscle fiber composition in human skeletal muscle biopsies is often performed using immunohistochemistry, a method that tends to be both time consuming, technically challenging, and complicated by limited availability of tissue. Here, we introduce quantitative reverse transcriptase polymerase chain reaction (qRT-PCR)-based Gene-family profiling (GeneFam) of myosin heavy chain (MyHC) mRNA expression as a high-throughput, sensitive, and reliable alternative. We show that GeneFam and immunohistochemistry result in similar disclosures of alterations in muscle fiber composition in biopsies from musculus vastus lateralis and musculus biceps brachii of previously untrained young women after 12 weeks of progressive strength training. The adaptations were evident as (a) consistent increases in MyHC2A abundance; (b) consistent decreases in MyHC2X abundance; and (c) consistently stable MyHC1 abundance, and were not found using traditional reference gene-based qRT-PCR analyses. Furthermore, muscle fiber composition found using each of the two approaches was correlated with each other (r = 0.50, 0.74, and 0.78 for MyHC1, A, and X, respectively), suggesting that GeneFam may be suitable for ranking of individual muscle phenotype, particularly for MyHC2 fibers. In summary, GeneFam of MyHC mRNA resulted in reliable assessment of alterations in muscle fiber composition in skeletal muscle of previously untrained women after 12 weeks of strength training.
在人体骨骼肌活检中,肌肉纤维组成的测定通常采用免疫组织化学方法,该方法往往既耗时、技术上具有挑战性,又因组织可用性有限而变得复杂。在此,我们引入基于定量逆转录聚合酶链反应(qRT-PCR)的肌球蛋白重链(MyHC)mRNA表达基因家族分析(GeneFam),作为一种高通量、灵敏且可靠的替代方法。我们发现,在未受过训练的年轻女性进行12周渐进性力量训练后,取自股外侧肌和肱二头肌的活检样本中,GeneFam和免疫组织化学在揭示肌肉纤维组成变化方面结果相似。这些适应性变化表现为:(a)MyHC2A丰度持续增加;(b)MyHC2X丰度持续降低;(c)MyHC1丰度持续稳定,而使用传统的基于参考基因的qRT-PCR分析未发现这些变化。此外,两种方法所测得的肌肉纤维组成彼此相关(MyHC1、A和X的相关系数分别为r = 0.50、0.74和0.78),这表明GeneFam可能适用于对个体肌肉表型进行排名,特别是对于MyHC2纤维。总之,MyHC mRNA的GeneFam分析能够可靠地评估未受过训练的女性在进行12周力量训练后骨骼肌中肌肉纤维组成的变化。