Real-time detection of α_(1A)-AR movement stimulated by phenylephrine in single living cells  

作　　者：Ning XU Zhang-yi LIANG Ming XU Ying-hua GUAN Qi-hua HE Qi-de HAN Xin-sheng ZHAO You-yi ZHANG 

机构地区：[1]Institute of Vascular Medicine, Peking University Third Hospital, and Key Laboratory of Molecular Cardiovascular Sciences, Ministry oJ Education, Beijing 100083, China [2]Beijing National Laboratory for Molecular Sciences, State Key Laboratory for Structural Chemistry of Unstable and Stable Species, and Department of Chemical Biology, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China [3]Center of Medical Analysis, Peking University, Health Science Center ,Beijing 100083, China

出　　处：《Acta Pharmacologica Sinica》2007年第6期796-802,共7页中国药理学报（英文版）

基　　金：grants from the National Natural Science Foundation of China(№ 20333010 and 30490172);the National Key Basic Research Science Foundation(NKBRSF)(№ G1999075305,2006C13503806)

摘　　要：Aim： To investigate the movement of α1A-adrenergic receptors（α1A-AR） stimulated by agonist, phenylephrine （PE）, and the dynamics of receptor movement in real time in single living cells with millisecond resolution. Methods： We labeled α1A-AR using the monoclonal, anti-FLAG （a kind of tag） antibody and Cy3-conju- gated goat anti-mouse IgG and recorded the trajectory of their transport process in living HEK293A cells stimulated by agonist, PE, and then analyzed their dynamic properties. Results： The specific detection of α1A-AR on the surface of living HEK293A-α1A cells was achieved, α1A-AR internalize under the stimulation of PE. After the cells were stimulated with PE for 20min, apparent colocalization was found between α1A-AR and F-actins. After 40 rain stimulation of PE, trajectories of approximate linear motion in HEK293A-α1A cells were recorded, and their velocity was calculated. Conclusion： The specific labeling method on the living cell surface provides a convenient means of real-time detection of the behavior of surface receptors. By this method we were able to specifically detect α1A-AR and record the behavior of individual particles of receptors with 50 ms exposure time in real time in single living cells.Aim： To investigate the movement of α1A-adrenergic receptors（α1A-AR） stimulated by agonist, phenylephrine （PE）, and the dynamics of receptor movement in real time in single living cells with millisecond resolution. Methods： We labeled α1A-AR using the monoclonal, anti-FLAG （a kind of tag） antibody and Cy3-conju- gated goat anti-mouse IgG and recorded the trajectory of their transport process in living HEK293A cells stimulated by agonist, PE, and then analyzed their dynamic properties. Results： The specific detection of α1A-AR on the surface of living HEK293A-α1A cells was achieved, α1A-AR internalize under the stimulation of PE. After the cells were stimulated with PE for 20min, apparent colocalization was found between α1A-AR and F-actins. After 40 rain stimulation of PE, trajectories of approximate linear motion in HEK293A-α1A cells were recorded, and their velocity was calculated. Conclusion： The specific labeling method on the living cell surface provides a convenient means of real-time detection of the behavior of surface receptors. By this method we were able to specifically detect α1A-AR and record the behavior of individual particles of receptors with 50 ms exposure time in real time in single living cells.

关 键 词：adrenergic receptor TRAJECTORY living cell 

分 类 号：R97[医药卫生—药品]