[The study of beta-2 adrenergic receptors gene transferred into cardiomyocytes by adenovirus vectors contain different promoter].
OBJECTIVE
To observe human beta(2) adrenergic receptors (beta(2)-AR) gene expression in cardiomyocytes transfected by adenoviral vector contain cmv or mlc-2v promoter.


METHODS
Two adenoviral vectors (Adcmv beta(2) AR, Admlc beta(2) AR) were constructed, in which beta(2)-AR gene is under control of either the ventricle-specific myosin light chain-2 (mlc-2v) or the cmv promoter. The cultured neonate rat ventricular myocytes were infected by two adenoviral vectors, and the beta(2)-AR expression of on cultured neonate rat ventricular myocytes and their ability to potentiate beta-adrenergic signaling were assessed.


RESULTS
The presence of beta(2)-AR mRNA was detected by RT-PCR, and the expression of the beta(2)-AR gene was demonstrated by protein immunoblots. According to a ligand binding assay, the density of beta-AR in the cardiomyocytes infected by Admlc beta(2) AR and Adcmv beta(2) AR was greater than that in the control. Moreover, the latter was higher than the former (234 +/- 6.4 fmol/mg protein vs 153 +/- 5.2 fmol/mg protein) (P < 0.01). It was demonstrated that beta(2)-AR gene significantly increased isoproterenol stimulated cAMP in cardiomyocytes.


CONCLUSIONS
The results above demonstrated that human beta(2)-AR gene was expressed in the cardiomyocytes infected by recombinant adenovirus contain cmv or mlc-2v promoter, but both is different.
Gao
W
W
Department of Senile Cardiology, General Hospital of PLA, Beijing 100853, China.
Li
X
X
Pei
X
X
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D009206:Myocardium / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D018343:Receptors, Adrenergic, beta-2; D000242:Cyclic AMP