@article{APS4816,
author = {Ning Xu and Zhang-yi Liang and Ming Xu and Ying-hua Guan and Qi-hua He and Qi-de Han and Xin-sheng Zhao and You-yi Zhang},
title = {Real-time detection of α1A-AR movement stimulated by phenylephrine in single living cells},
journal = {Acta Pharmacologica Sinica},
volume = {28},
number = {6},
year = {2016},
keywords = {},
abstract = {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-conjugated 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 20 min, apparent colocalization was found between α1A-AR and F-actins. After 40 min 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.},
issn = {1745-7254}, url = {http://www.chinaphar.com/article/view/4816}
}