1. Electrical field stimulation (EFS) of muscle strips in vitro elicited a tetrodotoxin (TTX)-sensitive biphasic contractile response consisting of a phasic component followed by a tonic one. 2. The amplitude of both components of the response was impaired by N omega-nitro-L-arginine and potentiated by sodium nitroprusside. Cystamine caused a reduction in amplitude of both phasic and tonic components of the response to EFS. Neither N omega-nitro-L-arginine, sodium nitroprusside, nor cystamine induced changes in the resting muscle tone, or in the contractile response to exogenous agonists ATP and noradrenaline (NA). 3. The nitric oxide scavenger, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, induced a reduction in amplitude of both components of the response to EFS. 4. These results reveal a facilitatory prejunctional modulatory role for nitric oxide in sympathetic neurotransmission in rat vas deferens. Endogenous nitric oxide released in the extracellular space is presumed to potentiate neurotransmission by acting at prejunctional level via cGMP.

1. The non-adrenergic non-cholinergic (NANC) inhibitory response to electrical field stimulation (EFS) in circular muscle from rat caecum was investigated using the single sucrose-gap technique. EFS with single pulses evoked hyperpolarization oral inhibitory function potential (IJP) of the membrane associated with muscular relaxation or with transient inhibition of spontaneous contractile activity. 2. The amplitude and the duration of the IJPs were enhanced by using train stimulation at increasing frequency. 3. Apamin (10(-7) M) reduced the amplitude of IJPs at all frequencies tested. 4. N omega-nitro-L-arginine methyl ester (L-NAME) (10(-4) M, 5 x 10(-4) M), but not D-NAME, caused a concentration dependent decrease in the amplitude of IJPs at all frequencies tested. L-Arginine (10(-3) M) prevented these effects. 5. L-NAME (5 x 10(-4) M) caused the disappearance of the apamin-resistant IJP-component, evoked by single pulse or by low frequency trains. 6. Sodium nitroprusside (SNP) (10(-4) M), a nitric oxide (NO) donor, induced hyperpolarization of membrane potential and muscular relaxation. SNP-induced effects were not affected by pretreatment of the muscle strips with effective concentrations of tetrodotoxin, apamin, and L-NAME. 7. P2-purinergic antagonists, reactive blue 2 (up to 5 x 10(-4) M) and suramin (up to 3 x 10(-4) M), failed to affect the evoked IJPs. 8. These results show that, in rat caecum, the NANC response to electrical stimulation is composed of two distinguishable components: an apamin-resistant and an apamin-sensitive component. NO or a related compound is mainly involved in the mediation of the apamin-resistant component, while ATP is not the mediator responsible for the apamin-sensitive component.

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1 This study was undertaken to analyse the involvement of ATP in non-adrenergic non- cholinergic (NANC) relaxation and possible interplay between nitrergic and purinergic systems in rat gastric fundus. 2 Experiments were performed in vitro on strips of longitudinal muscle from rat gastric fundus, recording the mechanical activity as changes in isometric force. In addition, NO release induced by different experimental conditions was assayed. 3 Under NANC conditions in serotonin-precontracted strips, electrical field stimulation (EFS) elicited a tetrodotoxin (TTX)-sensitive relaxation accompanied by nitric oxide (NO) release. This effect was antagonized by pretreatment with the NO synthase antagonist Nomega-nitro-L-arginine (L-NA) or by desensitization of purinergic receptors. Purinergic desensitization was also able to further antagonize the residual EFS-induced relaxation remaining after L-NA treatment. Exogenously applied NO [delivered as sodium nitroprusside (SNP)] or ATP (and related purines) induced concentration-dependent, TTX-insensitive relaxant responses. ATP also induced the release of NO. A reduction in the responses to ATP was observed in the presence of L-NA. In contrast, SNP-induced relaxation remained unchanged after desensitization of purinergic receptors. Finally, apamin, a blocker of the small conductance Ca2+ -dependent K+ channels, reduced the amplitude of the muscular relaxation evoked by either EFS, ATP or SNP. 4 In conclusion, this study provides evidence that in rat gastric fundus, ATP is one of the inhibitory transmitters released from NANC intramural neurones acting directly on the muscle, through receptors coupled to apamin-sensitive Ca2+ -dependent K+ channels and, indirectly, through the stimulation of NO production.

Pigeon oesophageal smooth muscle in vitro has spontaneous electromechanical activity. In the presence of atropine and guanethidine, electrical field stimulation evokes a transient TTX-sensitive response comprising inhibition of electric bursting activity and muscular relaxation. This NANC inhibitory response was analysed using the K+ channel blockers TEA and apamin, TEA perfusion (0.1-5 mM) induced a concentration-dependent reduction in amplitude of EFS-evoked relaxation. Responses to higher stimulation frequencies were more sensitive to TEA than those to lower ones. The maximum reduction in amplitude (29% of control) was obtained on 30 Hz EFS evoked responses during 5 mM TEA perfusion. In a similar way, apamin (0.01-10 microM) perfusion reduced NANC relaxation, up to 30% of control. These results suggest that in the pigeon oesophagus, NANC intramural neurons are responsible for muscular relaxation. We speculate that an increase in K+ conductance might be the main mechanism involved, although the residual response after K+ channel blockade indicates the existence of an additional ionic mechanism.