Expression of the tilapia prolactin receptor (tiPRL-R) has been characterized in the intestine of Oreochromis niloticus and the levels of both tiPRL-R transcripts and tiPRL binding sites have been further analyzed in this organ, as well as in gill and kidney, during adaptation of tilapia to a hyperosmotic environment. A single high-affinity binding site for tilapia PRL-I (tiPRL-I) was determined in full-length intestine by Scatchard analysis. A heterogeneous distribution of tiPRL-R was detected in this organ, with the posterior part always displaying a higher expression of both tiPRL-R transcript and tiPRL binding sites than the anterior and medial parts. Transfer of tilapia to brackish water (BW) led to an apparent increase in the specific binding of tiPRLs in intestine and gill even for long-term-adapted fish, whereas the high level of kidney tiPRL binding sites measured in control fish reared in fresh water was still detected in BW-adapted tilapia. There was no overall significant modification of tiPRL-R transcript levels in any organ during short-term or long-term adaptation, although a limited decrease occurred in the gill of BW-adapted fish, as shown earlier. Therefore, in O. niloticus adapted to BW, high and sustained levels of ti-PRL-R were observed in the three major osmoregulatory organs, gill, kidney, and intestine.