In euryhaline fish, prolactin (Prl) plays an essential role in freshwater (FW) acclimation. In the euryhaline and eurythermal Mozambique tilapia, Oreochromis mossambicus, Prl cells are model osmoreceptors, recently described to be thermosensitive. To investigate the effects of temperature on osmoreception, we incubated Prl cells of tilapia acclimated to either FW or seawater (SW) in different temperature (20, 26 and 32°C) and osmolality (280, 330 and 420 mOsm/kg) combinations for 6 h. Release of both Prl isoforms, Prl188 and Prl177, increased in hyposmotic media and were further augmented with a rise in temperature. Hyposmotically-induced release of Prl188 was inhibited at 20°C. In SW fish, mRNA expression of prl188 and prl177 showed direct and inverse relationships with temperature, respectively. In SW-acclimated tilapia Prl cells incubated in hyperosmotic media, Prl receptors, prlr1 and prlr2, and the stretch-activated Ca2+ channel, trpv4,were inhibited at 32°C, suggesting the presence of a cellular mechanism to compensate for elevated Prl release. Transcription factors, pou1f1, pou2f1b, creb3l1, cebpb, stat3, stat1a and nfat1c, known to regulate prl188 and prl177, were also downregulated at 32°C. Our findings provide evidence that osmoreception is modulated by temperature, and that both thermal and osmotic responses vary with acclimation salinity.
