Title Internalization and distribution of three alpha(1)-adrenoceptor subtypes in HEK293A cells before and after agonist stimulation
Authors Wang, Shu-yi
Song, Yao
Xu, Ming
He, Qi-hua
Han, Qi-de
Zhang, You-yi
Affiliation Peking Univ, Hosp 3, Inst Vasc Med, Beijing 100083, Peoples R China.
Minist Educ, Key Lab Mol Cardiovasc Sci, Beijing 100083, Peoples R China.
Peking Univ, Hlth Sci Ctr, Ctr Med Anal, Beijing 100083, Peoples R China.
Keywords adrenoceptor
agonist stimulation
confocal image
enzyme-linked immunosorbent assay
[H-3]-prazosin binding assay
internalization
subcellular distribution
subtype
PROTEIN-COUPLED RECEPTORS
ALPHA(1B)-ADRENERGIC RECEPTOR
ALPHA(1D)-ADRENERGIC RECEPTORS
SUBCELLULAR-LOCALIZATION
MEDIATED INTERNALIZATION
CELLULAR-LOCALIZATION
SMOOTH-MUSCLE
DESENSITIZATION
PHOSPHORYLATION
EXPRESSION
Issue Date 2007
Publisher 中国药理学报
Citation ACTA PHARMACOLOGICA SINICA.2007,28,(3),359-366.
Abstract Aim: To examine the subcellular distribution of the 3 alpha(1)-adrenoceptor (alpha(1)-AR) subtypes and their internalization and trafficking upon agonist stimulation in human embryonic kidney 293 A cells. Methods: Confocal real-time imaging, enzyme linked immunosorbent assay (ELISA) and whole cell [H-3]-prazosin binding assay were applied to detect the distribution and localization of the 3 alpha(1)-AR subtypes. Results: alpha(1A)-AR was found both on the cell surface and in the cytoplasm; alpha(1B)-AR, however, was predominantly detected on the cell surface, while alpha(1D)-AR was detected mainly in the intracellular compartments. After stimulation with phenylephrine, localization changes were detected by confocal microscopy for alpha(1A)- and alpha(1B)-AR, but the localization of alpha(1D)-AR were unaffected. Phenylephrine stimulation promoted a more rapid internalization of alpha(1B)-AR than alpha(1A)-AR. alpha(1D)-AR internalization was detected only by ELISA. Whole cell [H-3]-prazosin binding assay showed that alpha(1A)-AR functional receptors were detected both on the cell surface and in the cytoplasm; alpha(1B)-AR, however, were detected predominantly on the cell surface, while alpha(1D)-AR were detected mainly in intracellular compartments. Phenylephrine stimulation promoted internalization of alpha(1A)- and alpha(1B)-AR. Conclusion: Phenylephrine stimulation induced changes in the localization of the 3 alpha(1)-AR.
URI http://hdl.handle.net/20.500.11897/345492
ISSN 1671-4083
DOI 10.1111/j.1745-7254-2007-00509-x
Indexed SCI(E)
PubMed
中国科技核心期刊(ISTIC)
中国科学引文数据库(CSCD)
Appears in Collections: 第三医院

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