The antidepressant actions of the NMDA receptor antagonist ketamine require activity-dependent release of brain-derived neurotrophic factor (BDNF) in the medial prefrontal cortex (mPFC). Our recent study shows a similar requirement for vascular endothelial growth factor (VEGF). Previous studies have reported that both BDNF and VEGF activate TRPC3 and TRPC6 via diacylglycerol production, and that activation of TRPC3 and TRPC6 mediates the neurotrophic effects of BDNF. Here, we examined the roles of TRPC3 and TRPC6 in the mPFC in the antidepressant actions of ketamine in lipopolysaccharide (LPS)-induced depression model mice. In both the tail suspension and forced swim tests, the antidepressant-like actions of ketamine were significantly blocked by intra-mPFC infusion of the selective TRPC3 inhibitor Pyr3 or the selective TRPC6 inhibitor SAR7334. Intra-mPFC infusion of BDNF or VEGF produced antidepressant-like effects, and these antidepressant-like effects were also blocked by intra-mPFC infusion of Pyr3 or SAR7334. Moreover, both systemic and intra-mPFC injection of a TRPC3/TRPC6 activator GSK1702934A produced antidepressant-like effects. These treatments did not significantly affect locomotor activity in LPS-treated mice. The current results indicate that the activation of TRPC3 and TRPC6 as downstream targets of BDNF and VEGF in the mPFC plays an essential role in the antidepressant actions of ketamine. TRPC3 and TRPC6 may be promising targets for the development of novel, rapid-acting antidepressant agents.