Methods for Evaluation of Vascular Endothelial Cell Function with Transient Receptor Potential (TRP) Channel Drugs

Vascular endothelial transient potential (TRP) channels, located mostly on the plasma membrane of cells, are critical in regulatory and pathophysiological circumstances. The objective of this chapter is to describe several well-established approaches, ranging from function to molecular assays, to investigate the mechanistic role of TRP channels in vascular endothelial cells. We show experimental procedures and representative figures on the following methods: (1) Isolation and culture of vascular endothelial cells, (2) examination of electrophysiological activity of TRP channel by patch-clamping with whole-cell configuration and its function in vascular tone and blood flow by isometric tension and isobaric diameter measurements, and Laser Doppler flowmetry, (3) detection of TRP channel-mediated intracellular Ca²⁺ imaging by using fluorescent microscopy, and (4) determination of TRP channel interaction by coimmunoprecipitation, double immunofluorescence staining and Förster resonance energy transfer (FRET) detection.