1. A preparation of haemolysed rat erythrocytes (the haemolysate) blocked the relaxations of both the bovine retractor penis and the rat anococcygeus muscles in response to field stimulation of their non-adrenergic inhibitory nerves. The effective concentration range was 5-20 mul./ml. of haemolysate, equivalent to 0.25-1.0 mul./ml. of blood. The active principle in the haemolysate was a non-dialysable, heat-labile material of molecular weight between 50,000 and 100,000 daltons. If, as appeared probable, the active component of the haemolysate was oxyhaemoglobin, its effective blocking concentration was 0.5-2 muM.2. Haemolysate (5-20 mul./ml.) also blocked the relaxation of both the bovine retractor penis and the rat anococcygeus to the inhibitory factor extracted from the bovine retractor penis, an observation supporting the possibility that this inhibitory factor may be the transmitter released by the inhibitory nerves in these tissues. In the bovine retractor penis, haemolysate was also effective in blocking relaxations in response to sodium nitroprusside, but relaxations produced by prostaglandin E(1) or isobutylmethylxanthine were unchanged or only slightly reduced.3. In contrast, in the taenia of the guinea-pig caecum, haemolysate did not block the non-adrenergic inhibitory response to field stimulation, nor the relaxation produced by ATP, although it did block the relaxation produced by the inhibitory factor.4. In spiral strips of isolated rabbit aorta, haemolysate (10 mul./ml.) increased the contraction produced by noradrenaline and blocked the relaxation produced by the inhibitory factor. These were shown to be independent effects.5. Apamin, which blocked the relaxation of the taenia of the guinea-pig caecum elicited by either ATP or field stimulation of its non-adrenergic nerves, was without effect on relaxations of the bovine retractor penis or rat anococcygeus muscles in response to field stimulation of inhibitory nerves or to inhibitory factor.6. These differences in the blocking effects of apamin and haemolysate suggest either that the transmitter in the bovine retractor penis and rat anococcygeus differs from that in the guinea-pig taenia, or, if the transmitter is the same, then its mechanism of action differs.