We previously reported that social isolation promotes parental care in sexually naive male mice. This effect was blocked by exposure to chemosensory and auditory social signals derived from males in an adjacent compartment. In the present study, we examined whether the chemosensory signals detected in the vomeronasal organ (VNO) are involved in parental behavior by using mice deficient for a VNO-specific ion channel (Trpc2(-/-))and thus impaired in VNO-input signaling. We housed virgin homozygous Trpc2(-/-) and heterozygous Trpc2(+/-) males for 3 weeks during puberty (5-8 weeks old) alone or in groups of 3-5 males. At 8 weeks of age, the mice were placed with three pups in an observation cage and tested for parental behavior. The Trpc2(-/-) males housed under isolated conditions spent significantly longer in the vicinity of pups than did the Trpc2(-/-) males than had been group housed, whereas no isolation effect was observed in heterozygous Trpc2(+/-) males. Both Trpc2 knockout and isolation housing significantly increased the time males spent licking pups and crouching (arched back posture over pups to enable nursing), whereas only isolation housing increased the incidence of retrieval behavior. These results demonstrated that social signals transmitted not only through the VNO but also from other modalities, independent of each other, suppress the expression of parental behavior during puberty in sexually naive males. (C) 2016 Elsevier Inc All rights reserved.