A diacylglycerol photoswitching protocol for studying TRPC channel functions in mammalian cells and tissue slices
Trese Leinders‐Zufall, Ursula Storch, Michael Mederos y Schnitzler, Navin K. Ojha, Kohei Koike, Thomas Gudermann, Frank Zufall
Abstract
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).