Evidence for nitric oxide as an inhibitory neurotransmitter in rabbit isolated anococcygeus

The role of nitric oxide (NO) and ATP as putative inhibitory non-adrenergic, non-cholinergic (NANC) neurotransmitters was investigated in rabbit isolated anococcygeus after block of adrenergic and cholinergic responses, and raising tone with histamine. NANC nerve stimulation produced rapid relaxations which were completely abolished by tetrodotoxin. The magnitude of the NANC inhibitory responses was significantly reduced by the nitric oxide synthase inhibitors NG-nitro-L-arginine (NO-Arg) and NG-nitro-L-arginine methyl ester (L-NAME). This effect could be partially reversed by L-arginine but not by D-arginine. Oxyhaemoglobin inhibited NANC nerve responses and sodium nitroprusside mimicked the effects of NANC nerve stimulation. NO-Arg also reduced the magnitude of the inhibitory junction potentials recorded from the smooth muscle cells during NANC nerve stimulation. Exogenously applied ATP and adenosine each produced concentration dependent relaxations which were unaffected by the NO-synthase inhibitor NO-Arg. Relaxations to adenosine were virtually abolished by the P1 purinoceptor antagonist 8-(p-sulphophenyl)theophylline. Relaxations to ATP were also significantly reduced, indicating that part of the response to exogenous ATP is due to its breakdown to adenosine and subsequent action on P1 purinoceptors. Relaxations of the tissue to ATP and adenosine were unaffected by the P2 purinoceptor antagonist suramin. NANC nerve mediated responses were not significantly changed by either 8-(p-sulphophenyl)theophylline or suramin. These results suggest that NO is involved in inhibitory NANC neurotransmission in the rabbit isolated anococcygeus, but do not support a role for ATP as a NANC neurotransmitter in this tissue.