Pandya Kumar, Cowhig John, Brackhan Joe, Kim Hyung Suk, Hagaman John, Rojas Mauricio, Carter Charles W, Mao Lan, Rockman Howard A, Maeda Nobuyo, Smithies Oliver
Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC 27599, USA.
Proc Natl Acad Sci U S A. 2008 Sep 2;105(35):13063-8. doi: 10.1073/pnas.0805120105. Epub 2008 Aug 28.
To determine whether the expression of cardiac genes changes in a graded manner or by on/off switching when cardiac myocytes change genetic programs in living animals, we have studied two indicator genes that change their expression oppositely in mouse binucleate ventricular cardiomyocytes during development and in response to cardiac hypertrophy. One is a single-copy transgene controlled by an alpha-myosin heavy chain (aMHC) promoter and coding for CFP. The other is the endogenous beta-myosin heavy chain (bMHC) gene modified to code for a YFP-bMHC fusion protein. Using high-resolution confocal microscopy, we determined the expression of the two indicator genes in individual cardiomyocytes perinatally and after inducing cardiac hypertrophy by transverse aortic constriction. Our results provide strong evidence that the cardiac genes respond by switching their expression in an on/off rather than graded manner, and that responding genes within a single cell and within the two nuclei of cardiomyocytes do not necessarily switch concordantly.
为了确定在活体动物中,当心肌细胞改变基因程序时,心脏基因的表达是呈梯度变化还是通过开/关切换,我们研究了两个指示基因,它们在小鼠双核心室心肌细胞发育过程中以及对心脏肥大的反应中,表达呈相反变化。一个是由α-肌球蛋白重链(aMHC)启动子控制并编码CFP的单拷贝转基因。另一个是经过修饰以编码YFP-bMHC融合蛋白的内源性β-肌球蛋白重链(bMHC)基因。使用高分辨率共聚焦显微镜,我们在围产期以及通过横向主动脉缩窄诱导心脏肥大后,测定了单个心肌细胞中两个指示基因的表达。我们的结果提供了强有力的证据,表明心脏基因通过开/关方式而非梯度方式切换其表达来做出反应,并且单个细胞内以及心肌细胞的两个细胞核内的反应基因不一定会一致地切换。