1. The role of intraperitoneal osmoreceptors in hypothalamo-neurohypophyseal control was studied in urethane- or nembutal-anaesthetized rats. Plasma samples were taken for radioimmunoassay of arginine vasopressin, and the electrical activity of single supraoptic endocrine neurones and of the hypothalamo-neurohypophyseal tract were monitored during superfusion of the hepatic portal vein with hypo-, iso- and hypertonic solutions. 2. Plasma arginine vasopressin increased within 1 min following superfusion with 0.3-0.9 osmolal NaCl solutions in a dose-related manner from basal levels of 30 pg/ml, to 170 pg/ml. Prior superfusion with xylocaine or intravenous infusions of 800 micrograms atropine-methyl bromate abolished this response, although vasopressin was still released to nicotine in atropine-blocked rats. 3. Portal vein superfusions had no significant effects on arterial blood pressure, plasma osmolality and plasma Na concentrations. 4. Forty supraoptic neurones were antidromically activated from the neural lobe/stalk region. Superfusions of the portal vein with NaCl solutions (0.33-1.20 osmole/kg, 37 degrees C, 5-120 sec) stimulated seven out of eight phasically firing and eight out of twenty-four continuously firing neurones. One phasically active, ten continuously firing and four silent cells were not affected, and six continuously firing neurones were inhibited by the superfusions. 5. The amplitude decreases of antidromic compound action potentials in the hypothalamo-neurohypophyseal tract, reflecting an increase of the orthodromic nerve impulse traffic, ranged from 17 to 22% for superfusions with 1.2 osmolal NaCl or LiCl solutions, from 8 to 11% for 1.2 osmolal Na isethionate or choline Cl and from 3 to 9% for 1.2 osmolal glucose; there was no effect when 1.2 osmolal urea and isotonic or hypotonic NaCl solutions were applied. 6. Responses of the amplitude of compound action potentials to superfusions with 1.2 osmolal NaCl solutions or with 0.1 mumole ACh, but not to electrical stimulation of the portal vein or its superfusion with 1.2 osmolal KCl, were abolished by prior application of 0.3 mumole atropine sulphate. Prior superfusions with xylocaine abolished the responses to all stimuli above. 7. These results suggest that within the hepatic portal vein area there are osmosensitive receptor cells and/or nerve terminals which activate the hypothalamoneurohypophyseal system through a peripheral cholinergic mechanism.