,Internalization and distribution of three α1-adrenoceptor subtypes in HEK293A cells before and afte

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Aim: To examine the subcellular distribution of the 3 α1-adrenoceptor (α1-AR)subtypes and their intealization and trafficking upon agonist stimulation in hu-man embryonic kidney 293A cells. Methods: Confocal real-time imaging, enzyme linked immunosorbent assay (ELISA) and whole cell [3H]-prazosin binding assay were applied to detect the distribution and localization of the 3 α1-AR subtypes.Results: α1A-AR was found both on the cell surface and in the cytoplasm; α1B-AR, however, was predominantly detected on the cell surface, while α1D-AR was detected mainly in the intracellular compartments. After stimulation with phenylephrine, localization changes were detected by confocal microscopy for α1A-and α1B-AR, but the localization of α1D-AR were unaffected. Phenylephrine stimulation promoted a more rapid intealization of α1B-AR than α1A-AR. α1D-AR intealization was detected only by ELISA. Whole cell [3H]-prazosin binding assay showed that α1A-AR functional receptors were detected both on the cell surface and in the cytoplasm; α1B-AR, however, were detected predominantly on the cell surface, while α1D-AR were detected mainly in intracellular compartments.Phenylephrine stimulation promoted intealization of α1A- and α1B-AR.Conclusion: Phenylephrine stimulation induced changes in the localization of the 3 α1-AR.
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