Optical control of TRPV1 channels in vitro with tethered photopharmacology

Transient receptor potential vanilloid 1 (TRPV1) is a nonselective cation channel that is important for nociception and inflammatory pain, and is activated by a variety of nociceptive stimuli—including lipids such as capsaicin (CAP) and endocannabinoids. TRPV1’s role in physiological systems is often studied by activating it with externally perfused ligands; however, this approach is plagued by poor spatiotemporal resolution. Lipid agonists are insoluble in physiological buffers and can permeate membranes to accumulate non-selectively inside cells, where they have off-target effects. To increase the spatiotemporal precision with which we can activate lipids on cells and tissues, we previously developed Optically-Cleavable Targeted (OCT)-ligands, which use protein tags (SNAP-tags) to localize a photocaged ligand on a target cellular membrane. After enrichment, the active ligand is released on a flash of light to activate nearby receptors. In this work, we expand the scope of OCT-ligand technology to target TRPV1. We synthesize a probe, OCT-CAP, that tethers to membrane-bound SNAP-tags and releases a TRPV1 agonist when triggered by UV-A irradiation. Using Ca2+ imaging and electrophysiology in HEK293T cells expressing TRPV1, we demonstrate that OCT-CAP uncaging activates TRPV1 with superior spatiotemporal precision when compared to standard diffusible ligands or photocages. This study demonstrates the versatility of OCT-ligands to manipulate ion-channel targets, and we anticipate that these tools will find many applications in controlling lipid signaling pathways in various cells and tissues.