1 The relative contribution of the putative transmitters, nitric oxide (NO) and an apamin-sensitive factor, possibly ATP, to inhibitory responses evoked by electrical ®eld stimulation (EFS; 0.2 ± 5 Hz, 0.2 ms duration, supra-maximal voltage for 10 s) of non-adrenergic, non-cholinergic (NANC) nerves was investigated in the guinea-pig isolated taenia coli contracted with histamine (1 mM). 2 Peak relaxations to EFS (0.2 ± 5 Hz) were tetrodotoxin (1 mM)-sensitive, maximal at 0.2 Hz and completely resistant to the nitric oxide synthase inhibitor, N G -nitro-L-arginine (L-NOARG; 100 mM) in either the presence or absence of atropine (1 mM). Furthermore, the speci®c inhibitor of soluble guanylyl cyclase, 1H-[1,2,4] oxadiazolo [4,3-a] quinoxaline-1-one (ODQ; 10 mM), the cytochrome P 450 inhibitor and free radical generator, 7-ethoxyresoru®n (7-ER; 10 mM) and the NO scavenger, oxyhaemoglobin (HbO; 30 mM) had no e ect on EFS-induced relaxations alone and in combination with L-NOARG (100 mM).3 Maximum relaxation to the NO donor, sodium nitroprusside (SNP; 1 mM) was signi®cantly reduced by HbO (30 mM), abolished by 7-ER (10 mM) and ODQ (10 mM) but was una ected by apamin (0.1 mM), an inhibitor of small conductance Ca 2+ -activated K + channels. 4 The relaxation to EFS at 0.2 Hz was resistant to apamin but those to 0.5 and 5 Hz were signi®cantly reduced. EFS (0.2 ± 5 Hz)-evoked relaxations that persisted in the presence of apamin were further signi®cantly inhibited by L-NOARG (100 mM) or ODQ (10 mM), but not by HbO (30 mM) or 7-ER (10 mM). 5 ATP (1 ± 30 mM) produced concentration-dependent relaxations that were abolished by apamin (0.1 mM), una ected by ODQ (10 mM) but only signi®cantly reduced by L-NOARG (100 mM) at the lowest concentration of ATP (1 mM) used. 6 Nifedipine (0.3 mM), abolished contractions to 67 mM KCl, histamine (10 mM), endothelin-1 (0.03 mM), 5-hydroxytryptamine (5-HT; 10 mM) and the thromboxane-mimetic, 9-11-dideoxy-9a, 11a-methano-epoxy-prostaglandin F 2a (U46619; 0.1 mM). 7 The ®ndings of the present study suggest that NO is released during NANC nerve stimulation, but plays no role in NANC relaxations in the guinea-pig taenia coli unless the e ects of another apaminsensitive, nerve-derived hyperpolarizing factor (NDHF) are blocked. Thus, we propose that in this tissue, NO acts as a`backup' or redundant NANC nerve inhibitory transmitter and like NDHF mediates relaxation via hyperpolarization.