Neuropathic pain, a debilitating syndrome that occurs post-nerve damage, can lead to hypersensitivity in the peripheral and central nervous systems. The underlying mechanism is poorly understood, and currently available treatments remain inefficient. Therefore, elucidation of the molecular mechanisms which contribute to this syndrome could be important in developing more effective treatments regimens. Netrin, a laminin-related extracellular protein, was originally identified as an axonal guidance molecule in the embryonic spinal cord, and subsequent findings have demonstrated that Netrin plays an important role in cell survival, migration, axon branching, and synaptogenesis during development. However, the functional role of Netrin in the adult nervous system remains unknown. In the current study, we examined the role of Netrin-4 in neuropathic pain, and determined that this protein was expressed in neurons located in the inner lamina II of spinal cord dorsal horn. Behavioural analysis revealed that a lack of the Netrin-4 attenuated both mechanical and thermal allodynia in neuropathic pain. In contrast, intrathecal administration of Netrin-4 protein to naive rats induced mechanical allodynia, being mediated by Unc5B and tyrosine phosphatase SHP2. These findings suggest that transient suppression of Netrin-4-Unc5B signal proves as an efficient strategy for the treatment of allodynia after nerve injury. In addition, our findings provide evidence for the function of Netrin in adult nervous system.