We recently reported that Gly573Ser substitution of the transient receptor potential cation channel, subfamily V member 3 (TRPV3) caused hair loss in DS-Nh mice. To further elucidate the effects of this mutation on the development of the spontaneous hairless phenotype, we examined the temperature-response to epidermal sheets from DS-Nh and DS mice. It was indicated that the mutation was gain-of-function. We also performed genetic and histological analyses with both strain skins. DNA microarray data revealed that the levels of keratin-associated protein 16-1, 16-3, and 16-9 genes related to the anagen phase were decreased in the skins of DS-Nh mice compared with those of three days old DS mice. Histological analysis revealed that the anagen phase persisted in DS-Nh mice, and that the telogen phase was seen in DS but not DS-Nh mice at 21 days of age. Regulation of TRPV3 appears to be important for appropriate hair development in rodents.