The number of functional transient receptor potential vanilloid 1 (TRPV1) channels at the surface,especially at the peripheral terminals of primary sensory neurons, regulates heat sensitivity, and increased surface localization of TRPV1 s contributes to heat hyperalgesia.However, the mechanisms for regulating TRPV1 surface localization are largely unknown.Here, we show that cyclin-dependent kinase 5 (Cdk5), a new player in thermal pain sensation, positively regulates TRPV1 surface localization.We found that Cdk5 phosphorylates KIF13B at Thr-506.T506A mutation reduced the motor-cargo interaction and the cell-permeable TAT-T506 peptide, targeting to the Thr-506, decreased TRPV1 surface localization, demonstrating the essential role of Thr-506 phosphorylation in TRPV1 transport.Moreover, CFA injection-induced activation of Cdk5 increased the anterograde transport of TRPV1s, contributing to the development and possibly the maintenance of heat hyperalgesia, while intrathecal delivery of the TAT-T506 peptide alleviated CFA-induced heat hyperalgesia.Thus, Cdk5 regulation of TRPV1 membrane trafficking is a fundamental mechanism controlling the heat sensitivity of nociceptors and moderate inhibition of Thr-506 phosphorylation during inflammation might be helpful for the treatment of inflammatory thermal pain.