David A. Andersson

Mechanical and thermal cues stimulate a specialized group of sensory neurons that terminate in the skin. Three members of the transient receptor potential (TRP) family of channels are expressed in subsets of these neurons and are activated at distinct physiological temperatures. Here, we describe the cloning and characterization of a novel thermosensitive TRP channel. TRPV3 has a unique threshold: It is activated at innocuous (warm) temperatures and shows an increased response at noxious temperatures. TRPV3 is specifically expressed in keratinocytes; hence, skin cells are capable of detecting heat via molecules similar to those in heat-sensing neurons.

Transient receptor potential A1 (TRPA1) is expressed in a subset of nociceptive sensory neurons where it acts as a sensor for environmental irritants, including acrolein, and some pungent plant ingredients such as allyl isothiocyanate and cinnamaldehyde. These exogenous compounds activate TRPA1 by covalent modification of cysteine residues. We have used electrophysiological methods and measurements of intracellular calcium concentration ([Ca(2+)](i)) to show that TRPA1 is activated by several classes of endogenous thiol-reactive molecules. TRPA1 was activated by hydrogen peroxide (H(2)O(2); EC(50), 230 microM), by endogenously occurring alkenyl aldehydes (EC(50): 4-hydroxynonenal 19.9 microM, 4-oxo-nonenal 1.9 microM, 4-hydroxyhexenal 38.9 microM) and by the cyclopentenone prostaglandin, 15-deoxy-delta(12,14)-prostaglandin J(2) (15d-PGJ(2), EC(50): 5.6 microM). The effect of H(2)O(2) was reversed by treatment with dithiothreitol indicating that H(2)O(2) acts by promoting the formation of disulfide bonds whereas the actions of the alkenyl aldehydes and 15d-PGJ(2) were not reversed, suggesting that these agents form Michael adducts. H(2)O(2) and the naturally occurring alkenyl aldehydes and 15d-PGJ(2) acted on a subset of isolated rat and mouse sensory neurons [approximately 25% of rat dorsal root ganglion (DRG) and approximately 50% of nodose ganglion neurons] to evoke a depolarizing inward current and an increase in [Ca(2+)](i) in TRPA1 expressing neurons. The abilities of H(2)O(2), alkenyl aldehydes and 15d-PGJ(2) to raise [Ca(2+)](i) in mouse DRG neurons were greatly reduced in neurons from trpa1(-/-) mice. Furthermore, intraplantar injection of either H(2)O(2) or 15d-PGJ2 evoked a nocifensive/pain response in wild-type mice, but not in trpa1(-/-) mice. These data demonstrate that multiple agents produced during episodes of oxidative stress can activate TRPA1 expressed in sensory neurons.