Apamin- and nitric oxide-sensitive biphasic non-adrenergic non-cholinergicinhibitory junction potentials in the rat anococcygeus muscle

1. Changes in membrane potential following electrical field stimulation (EP S; 1, 2 and 5 pulses at 5 Hz, 0.5 ms duration, 60-80 V) of non-adrenergic n on-cholinergic (NANC) inhibitory nerves in the rat isolated anococcygeus mu scle were measured using standard intracellular recording techniques. Resti ng membrane potential ranged between -60 and -70 mV. 2. In the presence of guanethidine (30 mu M), atropine (1 mu M), propranolo l (1 mu M) and phentolamine (0.05 mu M) to establish NANC conditions, the m embrane potential depolarized to between -40 and -50 mV. Under these condit ions, EFS caused pulse-dependent, tetrodotoxin (1 mu M)-sensitive biphasic inhibitory junction potentials (IJPs) comprising a fast onset and time-to-p eak phase followed by a second, slower phase that delayed repolarization. T he duration of NANC IJPs ranged between 10 and 20 s. 3. Inhibition of small-conductance Ca2+-activated K+ channels with apamin ( 0.1 mu M) selectively blocked the first fast phase of the NANC IJP, whereas inhibitors of large-conductance Ca2+-activated K+ channels (charybdotoxin and iberiotoxin) and ATP-sensitive K+ channels (glibenclamide) all had no e ffect on NANC IJPs. 4. Both the nitric oxide synthase inhibitor N-G-nitro-L-arginine (L-NOARG; 100 mu M) and the inhibitor of soluble guanylate cyclase 1-H-oxodiazol-[1,2 ,4]-[4,3-a] quinoxaline-1-one (ODQ; 10 mu M) had no effect on the first fas t phase of the NANC IJP. Each treatment, however, markedly inhibited the sl ow phase with the duration of the IJP reduced to between 1 and 3 s. The L-N OARG-resistant fast phase of the NANC IJP was almost abolished by the subse quent addition of apamin (0.1 mu M). 5. In conclusion, the present study demonstrates unequivocal NANC nerve-med iated biphasic IJPs in the rat isolated anococcygeus. We propose that nitri c oxide (NO), via activation of cGMP-dependent K+ channels, and a non-NO in hibitory factor which activates apamin-sensitive K+ channels contribute to NANC nerve-evoked IJPs in the rat anococcygeus.