Abnormal cold sensitivity is a common feature of a range of neuropathies. In the murine somatosensory system, multiple aspects of cold sensitivity are dependent on TRPM8, both acutely and in response to peripheral nerve injury. The specialised nature of cold sensitive afferents and the restricted expression of TRPM8 render it an attractive target for the treatment of cold hypersensitivity. This current study examines the effect of a novel TRPM8 antagonist (M8-An) in naïve and spinal nerve ligated rats through behavioural and in vivo electrophysiological approaches. In vitro, M8-An inhibited icilin evoked Ca(2+) currents in HEK293 cells stably expressing human TRPM8 with an IC50 of 10.9 nM. In vivo, systemic M8-An transiently decreased core body temperature. Deep dorsal horn recordings were made in vivo from neurones innervating the hind paw. M8-An inhibited neuronal responses to innocuous and noxious cooling of the receptive field in spinal nerve ligated rats, but not in naive rats. No effect on neuronal responses to mechanical and heat stimulation was observed. In addition, M8-An also attenuated behavioural responses to cold but not mechanical stimulation after nerve ligation without affecting the uninjured contralateral response. The data presented here supports a contribution of TRPM8 to the pathophysiology of cold hypersensitivity in this model and highlights the potential of the pharmacological block of TRPM8 in alleviating the associated symptoms.