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Characterization of cAMP accumulation mediated by three alpha1-adrenoceptor subtypes in HEK293 cells
Yao Song1, Yan Huang, Er-Dan Dong
1Institute of Vascular Medicine, Third Hospital, Peking University, The Reference Laboratory of Ministry of Education on Molecular Cardiovasology, Beijing 100083, China.
Aim:
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.
Conclusion:
Each of three alpha1-AR subtypes can mediate cAMP accumulation in HEK293 cell line, and there are differences in pharmacological property.
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