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 |