Methods to Study the Effects of TRP Channel Drugs on Vascular Endothelial Cell Function

Transient receptor potential (TRP) channels are abundantly expressed in vascular endothelium where the channels play important functional roles. In this chapter, we describe some well-established approaches, from molecular detection to functional assays, to study the role of TRP channels in endothelial cells. Comprehensive step-by-step instructions and representative figures are provided on the following methods: (1) isolation and culture of vascular endothelial cells; (2) determination of the TRP channel interaction [co-immunoprecipitation, double immunofluorescence staining, chemical cross-linking, and F�rster resonance energy transfer (FRET) detection]; (3) detection of TRP channel-mediated intracellular Ca²⁺ changes using fluorescence microscopy; (4) determination of the electrophysiological properties by whole-cell patch clamp and examination of TRP channel function in vascular tone control using isometric tension measurement.