Evidence has accumulated that acetylcholine (ACh) is capable of acting on neurones at sites other than the subsynaptic membrane. It has been shown for instance that it can excite mammalian sensory nerve fibres in the skin and mesentery (Brown & Gray, 1948;Douglas & Gray, 1953), carotid sinus pressure receptors (Diamond, 1955) and C fibres carrying activity from cutaneous touch receptors (Douglas & Ritchie, 1960). Among the first to suggest that acetylcholine might excite post-ganglionic sympathetic fibres were Coon & Rothman (1940), who injected acetylcholine into the skin and obtained sympathetic effects which were prevented by ergotamine and by degeneration of the sympathetic supply to the area. Recently, Brandon & Rand (1961) and Daly & Scott (1961) have reinvestigated the sympathomimetic effect of acetylcholine on the spleen, previously described by Farber (1936). They showed that close arterial injection of acetylcholine into the spleen produced contraction of that organ and this effect could be prevented by reserpine, by adrenergic blocking agents, by hexamethonium and by degeneration of the sympathetic supply. It seemed possible that this sympathomimetic effect of acetylcholine might be due to its exciting the sympathetic post-ganglionic adrenergic C fibres, and experiments were carried out to test this hypothesis. Some of the results of these experiments have already been communicated to the Physiological Society (Ferry, 1962).
METHODSCats were anaesthetized with ethyl chloride and ether and then with intravenous chloralose, 80 mg/kg. The abdominal cavity was opened by a mid-line incision and the spleen and its arterial supply were separated from the stomach and omentum by tying and cutting the hepatic, left gastric and gastro-epiploic arteries. In some experiments the stomach was removed to gain better access to the arterial supply of the spleen.Acetylcholine solution was injected into the splenic artery through a polythene cannula placed in a suitable sidebranch and pointing toward the heart. The arteriesused for retrograde cannulation were the hepatic artery, the smaller of the two divisions of the splenic artery, or a small branch of the splenic artery just before it enters the spleen. In one experiment one of the gastro-epiploic arteries was used. The arrangement of the cannula was such that injections could be made without interrupting the natural blood supply during the injection.