Differential contribution of substance P and neurokinin a to spinal cord neurokinin-1 receptor signaling in the rat

Although the tachykinins substance P (SP) and neurokinin A (NKA) are coreleased from primary afferent nociceptors and act via neurokinin (NK) receptors, their differential effects in vivo are not known. Despite pharmacological evidence that NKA preferentially binds NK- 2 receptors, this receptor is not found in spinal cord neurons. Thus, in the present studies, we compared the extent to which SP and NKA contribute to spinal nociceptive processing via the NK- 1 receptor. We found that SP and NKA induce NK- 1 receptor internalization with identical dose dependence and induce increases in intracellular calcium at the same concentrations, suggesting that SP and NKA equally activate the NK- 1 receptor. We found, however, that the selective NK- 1 receptor antagonist GR 205171 blocked NKA but not SP- induced NK- 1 receptor internalization in the rat spinal cord in vivo and in embryonic day 19 rat spinal neurons in vitro. Using this selectivity of GR 205171 for NKA- induced NK- 1 receptor activation, we examined the relative contribution of SP and NKA to noxious stimulus-induced activation of spinal NK- 1 receptors. We estimate that NKA contributes to at least 50% of the NK- 1 receptor activation in lamina I. Under inflammatory conditions, all noxious stimulus- induced NK- 1 receptor internalization in deep dorsal horn neurons was blocked by GR 205171, suggesting that it is entirely NKA- mediated. Substance P- mediated NK- 1 receptor internalization was focused at the site of termination of stimulated nociceptors but NKA also activated NK- 1 receptors at more distant sites. We conclude that NKA not only targets the NK- 1 receptor but may be a predominant pronociceptive primary afferent neurotransmitter.