, 2009). TRPM3 is a member of the melastatin subfamily of TRP channels with limited homology to the heat-sensitive TRPV channels. It is expressed in a variety of neuronal and nonneuronal tissue (Grimm et al., 2003, Lee et al., 2003 and Oberwinkler and Philipp, 2007). The TRPM3 gene encodes for different TRPM3 isoforms due to alternative splicing and exon usage, leading to channels

with divergent pore and gating properties (Oberwinkler et al., 2005). The neurosteroid pregnenolone sulfate (PS) is currently the most potent known activator of TRPM3 (in buy Nintedanib casu, the α2 isoform [Wagner et al., 2008]), and PS-induced activation of TRPM3-like currents has been linked to vascular smooth muscle contraction and Ca2+-induced insulin release from pancreatic islets in vitro (Naylor et al., 2010 and Wagner et al., 2008). However, it is currently unclear whether PS-induced gating of TRPM3 is occurring in vivo, and the physiological roles of the channel remain largely unclear (Nilius and Voets, 2008). Previous studies demonstrating expression of TRPM3-encoding mRNA in sensory neurons (Lechner et al., 2009, Nealen et al., 2003 and Staaf et al., 2010), and PS-induced pain responses in mice (Ueda et al., 2001) encouraged us to investigate the possible role of TRPM3 in somatosensation and nociception. In this study, we found that TRPM3 is functionally Tofacitinib order expressed in a large subset of sensory neurons from

the dorsal root and trigeminal ganglia (DRG and TG), and accounts for the majority of PS responses in these cells. Intraplantar injection of PS evokes nocifensive responses in

however wild-type mice but not in Trpm3−/− mice, indicating that TRPM3 activation provokes pain. Moreover, we discovered that TRPM3 is activated by heat. Consequently, TRPM3-deficient mice exhibit clear deficits in their avoidance response to noxious heat, but not to noxious cold or mechanical stimuli. TRPM3-deficient mice also failed to develop heat hyperalgesia following an inflammatory challenge. Our results provide evidence that TRPM3 plays a previously unanticipated role in heat sensation and nociception. Using quantitative real time-PCR on freshly isolated mouse DRG and TG, we detected TRPM3 mRNA at levels comparable to that of known somatosensory TRP channels TRPA1, TRPM8, TRPV1, and TRPV2, and higher than that of the heat-activated TRPV3 and TRPV4 (Figure 1A). These results are in line with earlier studies showing significant TRPM3 mRNA levels in sensory neurons (Lechner et al., 2009, Nealen et al., 2003 and Staaf et al., 2010). In situ hybridization using a TRPM3-specific antisense RNA probe yielded a strong signal in the cell bodies of a large fraction of DRG and TG neurons, comparable to the signals obtained with a TRPV1-specific antisense RNA probe (Figure 1B and see Figure S1 available online). Visual inspection of different sections revealed a positive TRPM3 hybridization signal in 78% ± 6% of DRG neurons and 82% ± 5% TG neurons.