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

STAR Protoc. 2021 May 5;2(2):100527. doi: 10.1016/j.xpro.2021.100527. eCollection 2021 Jun 18.

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).

Keywords: Cell Biology; Microscopy; Molecular/Chemical Probes; Neuroscience.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Cytological Techniques / methods*
  • Diglycerides* / chemistry
  • Diglycerides* / pharmacology
  • Mice
  • Olfactory Receptor Neurons / cytology
  • Photochemical Processes*
  • Transient Receptor Potential Channels* / analysis
  • Transient Receptor Potential Channels* / chemistry
  • Transient Receptor Potential Channels* / drug effects
  • Transient Receptor Potential Channels* / metabolism
  • Vomeronasal Organ / cytology

Substances

  • 1,2-diacylglycerol
  • Diglycerides
  • Transient Receptor Potential Channels