In tilapia, there are two forms of prolactin (PRL) whose effects on sodium and chloride movements differ and depend on the living environment of the fish. To see whether different receptors or the same receptor mediates these different effects, we have characterized the specific binding of both forms of tilapia (ti)PRL in two osmoregulatory organs, the gill and kidney. Two recombinant tiPRLs were used for this analysis. The recombinant hormones had the same properties as the native hormones in a tilapia gill radioreceptor assay. Specific binding to gill and kidney membranes was increased by optimizing the quality of the tissue preparations (physiological state of fish, membrane preparation) and the incubation conditions (pH, salt concentrations, temperature, time). Under these optimized conditions, we detected only one class of high affinity PRL receptor in gill and kidney. Its binding affinity was higher for tiPRLI than for tiPRLII in both gill and kidney (for tiPRLI the respective affinity values were 2.9 and 2.3 x 10(10) per M, for tiPRLII they were 1.9 and 0.5 x 10(10) per M). In competition studies, tiPRLI was more potent, followed by tiPRLII and ovine (o)PRL. tiGH and oGH did not significantly displace either tiPRL. The receptor we have characterized thus recognizes quite specifically both tiPRLs.