A diacylglycerol photoswitching protocol for studying TRPC channel functions in mammalian cells and tissue slices

Abstract

Summary: Small molecular probes designed for photopharmacology and opto-chemogenetics are rapidly gaining widespread recognition for investigations of transient receptor potential canonical (TRPC) channels. This protocol describes the use of three photoswitchable diacylglycerol analogs—PhoDAG-1, PhoDAG-3, and OptoDArG—for ultrarapid activation and deactivation of native TRPC2 channels in mouse vomeronasal sensory neurons and olfactory type B cells, as well as heterologously expressed human TRPC6 channels. Photoconversion can be achieved in mammalian tissue slices and enables all-optical stimulation and shutoff of TRPC channels.For complete details on the use and execution of this protocol, please refer to Leinders-Zufall et al. (2018).

Keywords

• Cell Biology
• Microscopy
• Neuroscience
• Molecular/Chemical Probes