Cardiovascular Development Research Program, Children's Hospital of Pittsburgh of University of Pittsburgh Medical Center, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
PLoS One. 2012;7(7):e40725. doi: 10.1371/journal.pone.0040725. Epub 2012 Jul 10.
Skeletal muscle derived stem cells (MDSCs) transplanted into injured myocardium can differentiate into fast skeletal muscle specific myosin heavy chain (sk-fMHC) and cardiac specific troponin-I (cTn-I) positive cells sustaining recipient myocardial function. We have recently found that MDSCs differentiate into a cardiomyocyte phenotype within a three-dimensional gel bioreactor. It is generally accepted that terminally differentiated myocardium or skeletal muscle only express cTn-I or sk-fMHC, respectively. Studies have shown the presence of non-cardiac muscle proteins in the developing myocardium or cardiac proteins in pathological skeletal muscle. In the current study, we tested the hypothesis that normal developing myocardium and skeletal muscle transiently share both sk-fMHC and cTn-I proteins. Immunohistochemistry, western blot, and RT-PCR analyses were carried out in embryonic day 13 (ED13) and 20 (ED20), neonatal day 0 (ND0) and 4 (ND4), postnatal day 10 (PND10), and 8 week-old adult female Lewis rat ventricular myocardium and gastrocnemius muscle. Confocal laser microscopy revealed that sk-fMHC was expressed as a typical striated muscle pattern within ED13 ventricular myocardium, and the striated sk-fMHC expression was lost by ND4 and became negative in adult myocardium. cTn-I was not expressed as a typical striated muscle pattern throughout the myocardium until PND10. Western blot and RT-PCR analyses revealed that gene and protein expression patterns of cardiac and skeletal muscle transcription factors and sk-fMHC within ventricular myocardium and skeletal muscle were similar at ED20, and the expression patterns became cardiac or skeletal muscle specific during postnatal development. These findings provide new insight into cardiac muscle development and highlight previously unknown common developmental features of cardiac and skeletal muscle.
骨骼肌源干细胞(MDSCs)移植到损伤的心肌中可以分化为快速骨骼肌特异性肌球蛋白重链(sk-fMHC)和心脏特异性肌钙蛋白 I(cTn-I)阳性细胞,维持受者心肌功能。我们最近发现 MDSCs 在三维凝胶生物反应器中分化为心肌细胞表型。一般认为,终末分化的心肌或骨骼肌分别仅表达 cTn-I 或 sk-fMHC。研究表明,在发育中的心肌中存在非心肌蛋白,或在病理性骨骼肌中存在心脏蛋白。在本研究中,我们检验了这样一个假设,即正常发育中的心肌和骨骼肌会短暂地共同表达 sk-fMHC 和 cTn-I 蛋白。在胚胎第 13 天(ED13)和第 20 天(ED20)、新生第 0 天(ND0)和第 4 天(ND4)、出生后第 10 天(PND10)和 8 周龄成年雌性 Lewis 大鼠心室心肌和比目鱼肌中进行了免疫组织化学、western blot 和 RT-PCR 分析。共聚焦激光显微镜显示,sk-fMHC 在 ED13 心室心肌中以典型的横纹肌模式表达,并且在 ND4 时横纹 sk-fMHC 表达消失,在成年心肌中呈阴性。cTn-I 直到 PND10 才以典型的横纹肌模式表达。Western blot 和 RT-PCR 分析显示,在 ED20 时,心室心肌和骨骼肌中的心脏和骨骼肌转录因子和 sk-fMHC 的基因和蛋白表达模式相似,在出生后发育过程中表达模式变得具有心脏或骨骼肌特异性。这些发现为心肌发育提供了新的见解,并强调了心脏和骨骼肌之前未知的共同发育特征。