EFFECT OF 7-NITRO INDAZOLE ON NEUROTRANSMISSION IN THE RAT VAS-DEFERENS - MECHANISMS UNRELATED TO INHIBITION OF NITRIC-OXIDE SYNTHASE

1 The effect of the nitric oxide synthase (NOS) inhibitor, 7-nitro indazole (7-NI), on sympathetic and purinergic neurotransmission in the rat isolated vas deferens preparation has been studied. 2 7-NI (50-200 mu M) caused a dose- and frequency-dependent inhibition of the phasic (predominantly purinergic) contractile response of the rat vas deferens to electrical (field) stimulation (100 V, 0.5 ms). Greatest inhibition occurred at lower frequencies of stimulation (0.1-10 Hz). The sustained tonic contractile response (predominantly noradrenergic) was inhibited only at a high frequency of stimulation (60 Hz) and only at the highest concentration of 7-NI studied (200 mu M). 3 7-NI (100 mu M) significantly reduced the contractile response of the vas deferens to exogenous ATP (20 mu M-5 mM) and the stable P-2X purinoceptor agonist, alpha,beta-methylene ATP (2.5 and 25.0 mu M) but was without effect on contractions due to noradrenaline (0.1-50 mu M) indicating a lack of antagonist effect on post-junctional alpha(1) adrenoceptors. 4 The effect of 7-NI (100 mu M) on the phasic contractile response to field stimulation (0.1 and 2.0 Hz) was unaffected by preincubation of preparations with yohimbine (1.0 mu M) or propranolol (0.01-10.0 mu M) indicating the absence of involvement of alpha(2)- or beta-adrenoceptors in this response. 5 7-NI (50-600 mu M) caused dose-related inhibition of contractions elicited by addition of a depolarizing concentration of KCl (64 mM). 6 The effect of 7-NI (100 mu M) on the phasic contractile response to held stimulation (0.1 and 2.0 Hz) was unaffected by preincubation of preparations with L-arginine (1 mM). Neither L-arginine (1 mM) nor N-G nitro L-arginine methyl ester (L-NAME, 100 mu M) affected the response of the vas deferens to field stimulation at 0.1 or 2.0 Hz. Nitric oxide synthase (NOS) enzyme activity, measured as the conversion of [H-3]-L-arginine to [H-3]-citrulline, was not detectable in rat vas deferens homogenates. 7 7-NI preferentially inhibits the purinergic component of the response of the rat vas deferens to held stimulation. The mechanism of action of 7-NI is not known but is not related to NOS inhibition. It seems likely that 7-NI combines an antagonist action at smooth muscle cell P-2X-purinoceptors with the ability to inhibit the cellular influx of calcium ions. Although these hitherto unrecorded effects of 7-NI occur at relatively high concentrations, the effects described may contribute to the pharmacological effects of this NOS inhibitor.