A-317491

Nociceptive primary afferents have the capacity to induce a state of increased excitability in the dorsal horn neurons of the spinal cord. It is well accepted that capsaicin-sensitive C-fibers transduce noxious stimulation and acute pain and that capsaicin-insensitive A beta-fibers are responsible for touch and innocuous sensation. It has been reported that the intrathecal (i.t.) administration of prostaglandin F(2 alpha) (PGF(2 alpha)) and ATP induces mechanical allodynia via the capsaicin-insensitive primary afferent pathway. In the present study, we investigated the interaction of purinoceptor P2X and the PGF(2 alpha) receptor (FP) in the induction of allodynia by use of mice lacking FP (FP(-/-)). Both PGF(2 alpha) and the P2X receptor agonist alphabeta-methylene ATP administered i.t. strongly induced allodynia for 50 min by tactile stimuli to the flank of mice. The allodynia induced by alphabeta-methylene ATP, but not that by PGF(2 alpha), was suppressed by simultaneous i.t. administration of P2X receptor antagonists pyridoxalphosphate-6-azophenyl-2,4-disulphonic acid and A-317491. In contrast, the allodynia induced by alphabeta-methylene ATP as well as that by PGF(2 alpha) was not observed in FP(-/-) mice. Immunostaining of beta-galactosidase, a reporter knocked into the endogenous FP locus in FP(-/-) mice, showed that the FP receptor was co-localized with P2X(2) and P2X(3) receptors in neurons of the spinal cord. alphabeta-Methylene ATP evoked a transient or sustained [Ca(2+)](i) increase in most of the PGF(2 alpha)-responsive cells in the deeper layer of the spinal cord, and the alphabeta-methylene ATP-evoked increase was blocked by the FP receptor antagonist AL-8810 in two-thirds of the cells. Neither PGF(2 alpha) nor alphabeta-methylene ATP induced the activation of spinal microglia. The present study demonstrates that the alphabeta-methylene ATP-evoked allodynia is mediated by the FP receptor, possibly via the functional coupling between the activation of P2X(2/3) receptors on the central terminal of capsaicin-insensitive fibers and FP receptors on spinal neurons.