, 2011). Having established the essential role of DEG-1, next we sought to determine how missense mutations in the DEG-1 protein affect MRCs by recording from deg-1(u506u679) mutants. This mutant allele was recovered

in a screen for suppressors of deg-1(u506)-induced necrotic cell death and encodes two point mutations ( García-Añoveros et al., 1995): an alanine to threonine change in the extracellular domain (A393T) that causes cell death when present alone and a glycine to arginine change in the conserved second transmembrane domain (G710R) that suppresses the A393T-induced cell death. We chose to study this allele because a change in the equivalent glycine residue of MEC-4(G716D) or MEC-10(G676R) alters the reversal potential and ion selectivity of MRCs recorded in PLM neurons ( Figure 4A; O’Hagan et al., 2005). If DEG-1 Alectinib cell line is a pore forming subunit of the MeT channel then the G710R mutation should

shift the reversal potential of MRCs in ASH. We VX-809 mouse tested this prediction by recording MRCs in deg-1(u506u679). Mechanoreceptor currents in u506u679 mutants were smaller than in wild-type ( Figures 4B and 4C) but larger than in deg-1 deletion mutants ( Table 1), suggesting that this allele is not null. Nevertheless, the effect of u506u679 on MRC amplitude is sufficient to induce a modest decrease in the ability of animals to respond to nose touch ( Figure 4D). Unlike wild-type MRCs, which have an estimated reversal potential of more than +100 mV in control saline, u506u679 MRCs reverse polarity near 0 mV ( Figure 4E). Thus, u506u679 alters the ion selectivity of MRCs in vivo. We note that the reversal potential of this mutant is different than that measured for deg-1 null mutants, supporting the idea that u506u679 is not a null allele of deg-1. We do not know whether the effect of u506u679 on ion selectivity is due to the extracellular A393T mutation, the G710R mutation in the second transmembrane domain, or both.

However, since the G710R mutation in DEG-1 affects the residue equivalent to the one mutated in mec-4(u2) [G716D] and mec-10(u20) [G676R] that alters the reversal potential of MRCs in PLM, it seems likely ALOX15 that this point mutation accounts for the change in selectivity. Regardless of whether the change in selectivity depends on one or both point mutations, this finding demonstrates DEG-1 is a pore-forming subunit of a channel that is critical for generating mechanoreceptor currents in ASH. The osm-9 and ocr-2 genes encode TRPV channel proteins coexpressed in ASH and required for ASH-mediated responses to noxious physical and chemical stimuli ( Colbert et al., 1997 and Tobin et al., 2002). Loss of osm-9 inhibits nose touch-evoked calcium transients in ASH ( Hilliard et al., 2005), supporting the idea that TRPV proteins form sensory mechanotransduction channels in ASH and elsewhere. Until now, this idea has not been tested directly.