Characterization of cAMP accumulation mediated by three alpha1-adrenoceptor subtypes in HEK293 cells.
OBJECTIVE
To investigate the characterization of cAMP response mediated by alpha1-adrenoceptor (alpha1-AR) subtypes in HEK293 cells.


METHODS
(1) Full-length cDNA encoding three alpha1-AR subtypes were transfected into HEK293 cells by the calcium phosphate precipitation method, respectively. (2) The densities of alpha1-AR subtypes expressed in HEK293 cells were measured by radioligand binding assay. (3) cAMP accumulation was measured by [3H]adenine prelabeling method.


RESULTS
(1) Activation of each of three subtypes resulted in an increase of cAMP accumulation in HEK293 cells in a dose-dependent manner, which was inhibited by selective alpha1-AR antagonist prazosin. (2) Comparing the pharmacological property, the maximal responses of alpha1A-AR to agonists were the most potent, while the sensitivity of alpha1-AR subtypes to norepinephrine (NE) was the highest.


CONCLUSIONS
Each of three alpha1-AR subtypes can mediate cAMP accumulation in HEK293 cell line, and there are differences in pharmacological property.
Song
Yao
Y
Institute of Vascular Medicine, Third Hospital, Peking University, The Reference Laboratory of Ministry of Education on Molecular Cardiovasology, Beijing 100083, China.
Huang
Yan
Y
Dong
Er-Dan
ED
Han
Qi-De
QD
Zhang
You-Yi
YY
D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D004622:Embryo, Mammalian; D006801:Humans; D007668:Kidney / Q000166:cytology*; D009638:Norepinephrine / Q000494:pharmacology; D018340:Receptors, Adrenergic, alpha-1 / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
C547713:ADRA1A protein, human; C547716:ADRA1B protein, human; D018076:DNA, Complementary; D018340:Receptors, Adrenergic, alpha-1; D000242:Cyclic AMP; D009638:Norepinephrine