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Abstract Animals must sense and acclimatize to environmental temperatures for survival, yet their thermosensing mechanisms other than transient receptor potential (TRP) channels remain poorly understood. We identify a trimeric G protein-coupled receptor (GPCR), SRH-40, which confers thermosensitivity in sensory neurons regulating temperature acclimatization in Caenorhabditis elegans. Systematic knockdown of 1000 GPCRs by RNAi reveals GPCRs involved in temperature acclimatization, among which srh-40 is highly expressed in the ADL sensory neuron, a temperature-responsive chemosensory neuron, where TRP channels act as accessorial thermoreceptors. In vivo Ca2+ imaging demonstrates that an srh-40 mutation reduced the temperature sensitivity of ADL, resulting in supranormal temperature acclimatization. Ectopically expressing SRH-40 in a non-warmth-sensing gustatory neuron confers temperature responses. Moreover, temperature-dependent SRH-40 activation is reconstituted in Drosophila S2R+ cells. Overall, SRH-40 may be involved in thermosensory signaling underlying temperature acclimatization. We propose a dual thermosensing machinery through a GPCR and TRP channels in a single sensory neuron.

Transient receptor potential (TRP) channels are primary sensory molecules in animals and are involved in detecting a diverse range of physical and chemical cues in the environments. Considering the crucial role of TRPA1 channels in nocifensive behaviors and aversive responses across various insect species, activators of TRPA1 are promising candidates for insect pest control. In this study, we demonstrate that 2-methylthiazoline (2MT), an artificial volatile thiazoline compound originally identified as a stimulant for mouse TRPA1, can be utilized as a novel repellent for fruit flies, Drosophila melanogaster. We observed that 2MT induced strong, dose-dependent avoidance behaviors in adult males, regardless of their feeding states, as well as egg laying behavior in females. These aversive responses were mediated by contact chemosensation via TrpA1 and olfaction via odorant receptors. Knocking down TrpA1 revealed the essential roles of bitter taste neurons and nociceptive neurons in the legs and labellum. Furthermore, among five isoforms, TrpA1-C and TrpA1-D exclusively contributed to the aversiveness of 2MT. We also discovered that these isoforms were directly activated by 2MT through covalent modification of evolutionarily conserved cysteine residues. In conclusion, we have identified 2MT as a stimulant for multiple sensory pathways, triggering aversive behaviors in fruit flies. We propose that 2MT and related chemicals may serve as potential resources for developing novel insect repellents.

Drosophila third-instar larvae exhibit changes in their behavioral responses to gravity and food as they transition from feeding to wandering stages. Using a thermal gradient encompassing the comfortable range (18°C to 28°C), we found that third-instar larvae exhibit a dramatic shift in thermal preference. Early third-instar larvae prefer 24°C, which switches to increasingly stronger biases for 18°C–19°C in mid- and late-third-instar larvae. Mutations eliminating either of two rhodopsins, Rh5 and Rh6, wiped out these age-dependent changes in thermal preference. In larvae, Rh5 and Rh6 are thought to function exclusively in the light-sensing Bolwig organ. However, the Bolwig organ was dispensable for the thermal preference. Rather, Rh5 and Rh6 were required in trpA1-expressing neurons in the brain, ventral nerve cord, and body wall. Because Rh1 contributes to thermal selection in the comfortable range during the early to mid-third-instar stage, fine thermal discrimination depends on multiple rhodopsins.

The skin barrier function is indispensable for terrestrial animals to avoid dehydration. The function is achieved by a hydrophobic cornified layer consisting of dead keratinocytes and lipids, and by an intercellular junction barrier formed among differentiated keratinocytes. A recent report demonstrated that TRPV4, one of the temperature-sensitive cation channels, contributes to the formation and maintenance of the intercellular junction-dependent barrier in the skin. TRPV4 associates with the E-cadherin complex via β-catenin, and thereby participates in the promotion of cell-cell junction development. TRPV4 allows influx of Ca2+ ions from the extracellular space at physiological skin temperatures. The Ca2+ influx induces Rho activation and promotes actin fiber organization and junction formation, thereby augmenting barrier integrity. Indeed, the intercellular junction structures and the skin barrier function were impaired in TRPV4-deficeint mice. This novel role of TRPV4 in keratinocytes may explain the significant correlation between temperature and the condition of skin.