Driesen Ronald B, Verheyen Fons K, Schaart Gert, de Mazière Ann, Viebahn Christoph, Prinzen Frits W, Lenders Marie-Hélène, Debie Wiel, Totzeck Andreas, Borgers Marcel, Ramaekers Frans C S
Department of Molecular Cell Biology, School for Cardiovascular Diseases (CARIM), University of Maastricht, Maastricht, The Netherlands.
Cardiovasc Pathol. 2009 Jan-Feb;18(1):19-27. doi: 10.1016/j.carpath.2007.12.008. Epub 2008 Feb 20.
Cardiotin expression is observed in adult cardiac tissue. In the present study, we provide evidence for the specific localization of cardiotin in cardiac mitochondria and for its down-regulation during adaptive remodeling (dedifferentiation) of cardiomyocytes.
Immunocytochemistry was used to study cardiotin localization in adult rabbit papillary muscle, in late-stage embryonic rabbit left ventricular tissue, and in left ventricle samples of rabbits suffering from pressure and volume overload. Western blot analysis of cardiotin was performed in purified pig heart mitochondrial fractions. Cardiotin expression was monitored in vitro in isolated adult rat and rabbit left ventricular cardiomyocytes.
Western blot analysis revealed the presence of cardiotin in the mitochondrial fractions of pig heart. Immunoelectron microscopy confirmed the presence of cardiotin in cardiac mitochondria of normal adult rabbits both in vivo and in vitro. Quantification of the localization of immunogold particles suggests an association of cardiotin with the mitochondrial inner membrane. Cardiotin expression is initiated in late-stage embryonic rabbit heart, whereas in adult ventricular tissue cardiotin clearly stained longitudinal arrays of mitochondria. Pressure- and volume-overloaded myocardium showed a reduction in cardiotin expression in dispersed local myocardial areas. Cell cultures of adult cardiomyocytes showed a gradual loss in cardiotin expression in parallel with a sarcomeric remodeling.
Our results demonstrate the specific localization of cardiotin in adult cardiomyocyte mitochondria and propose its use as an early marker for cardiomyocyte adaptive remodeling and dedifferentiation.
在成年心脏组织中观察到心脏素的表达。在本研究中,我们提供了证据,证明心脏素在心脏线粒体中的特定定位以及在心肌细胞适应性重塑(去分化)过程中其表达下调。
采用免疫细胞化学方法研究心脏素在成年兔乳头肌、晚期胚胎兔左心室组织以及压力和容量超负荷兔的左心室样本中的定位。对纯化的猪心脏线粒体组分进行心脏素的蛋白质印迹分析。在体外对分离的成年大鼠和兔左心室心肌细胞中的心脏素表达进行监测。
蛋白质印迹分析显示猪心脏线粒体组分中存在心脏素。免疫电子显微镜证实成年正常兔心脏线粒体在体内和体外均存在心脏素。免疫金颗粒定位的定量分析表明心脏素与线粒体内膜有关联。心脏素表达在晚期胚胎兔心脏中开始,而在成年心室组织中,心脏素清晰地染色线粒体的纵向排列。压力和容量超负荷的心肌在局部心肌区域分散处显示心脏素表达减少。成年心肌细胞的细胞培养显示心脏素表达逐渐丧失,同时伴有肌节重塑。
我们的结果证明了心脏素在成年心肌细胞线粒体中的特定定位,并提出将其用作心肌细胞适应性重塑和去分化的早期标志物。