Cell-specific expression of SERCA, the exogenous Ca~(2+) transport ATPase,in cardiac myocytes

摘要

We evaluated various constructs to obtain cell-specific expression of the sarco(endo)plasmic reticulum Ca~(2+)-ATPase (SERCA) gene in cardiac myocytes after cDNA transfer by means of transfections or infections with adenovi-rus vectors. Expression of exogenous enhanced green fluorescent protein (EGFP) and SERCA genes was studied in cultured chicken embryo and neonatal rat cardiac myocytes, skeletal and smooth muscle cells, fibroblasts, and hepatocytes. Whereas the cytomegalo-virus (CMV) promoter yielded high levels of protein expression in all cells studied, cardiac troponin T (cTnT) promoter segments demonstrated high specificity for cardiac myocytes. Their efficiency for protein expression was lower than that of the CMV promoter, but higher than that of cardiac myosin light chain or beta-myosin heavy chain promoter segments. A double virus system for Cre-dependent expression under control of the CMV promoter and Cre expression under control of a cardiac-specific promoter yielded high protein levels in cardiac myocytes, but only partial cell specificity due to significant Cre expression in hepatocytes.