Immune cells, epithelia and blood cells generate high levels of sphingosine-1-phosphate (S1P) in inflamed tissue. To date, it is not known if S1P affects nociceptive neurons, thereby contributing to the generation of inflammatory pain. Using a combination of several methods including behavioral tests (Hargreaves test), indirect immune fluorescence, quantative PCR and electrophysiological methods we found that the S1P₁ receptor for S1P is expressed in subpopulations of sensory neurons including nociceptors. Both, S1P and the agonist SEW2871 at the S1P₁ receptor induced hypersensitivity to noxious thermal stimulation invivo and in vitro. S1P-induced hypersensitivity was strongly attenuated in mice lacking TRPV1 channels. S1P and inflammation-induced hypersensitivity were significantly reduced in mice with a conditional nociceptor-specific deletion of the S1P₁ receptor. Our data show that neuronally expressed S1P₁ receptors play a significant role in regulating nociceptor function and that S1P/S1P₁ signaling may be a key player in the onset of thermal hypersensitivity and hyperalgesia associated with inflammation.