Chronically cannulated seawater (SW)-adapted flounder (Platichthys flesus) were used unanesthetized and unrestrained in an experimental series that acutely manipulated blood volume and plasma osmolality to determine their influence on plasma arginine vasotocin (AVT) concentrations. Immunoreactive AVT was measured using a radioimmunoassay validated for flounder and other teleosts. After hemorrhage-induced hypovolemia or hypervolemia produced by saline infusion, no major changes in plasma AVT concentrations were detected. Raising plasma osmolality by intraperitoneal injection of 1 M NaCl compared with control 150 mM NaCl-injected fish (329.4 +/- 3.4 vs. 320.4 +/- 3.0 mosmol/kgH2O, P < 0.05) produced an increase in plasma AVT concentration (6.7 +/- 1.2 vs. 4.2 +/- 0.2 fmol/ml, P < 0.05). In a separate study, plasma composition in a large number of uncannulated SW-adapted flounder was determined. This demonstrated a positive linear relationship between the natural variation in plasma AVT concentrations and plasma osmolality and Na+ and Cl- concentrations observed between fish. These data indicate that AVT secretion in SW-adapted flounder is closely related and perhaps directly sensitive to changes in plasma tonicity.