clc-2c is regulated by salinity, prolactin and extracellular osmolality in tilapia gill.

Teleosts inhabiting fresh water (FW) depend upon ion-absorptive ionocytes to counteract diffusive ion losses to the external environment. A Clc Cl- channel family member, Clc-2c, was identified as a conduit for basolateral Cl- transport by Na+/Cl- cotransporter 2 (Ncc2)-expressing ionocytes in stenohaline zebrafish (Danio rerio). It is unresolved whether Clc-2c/clc-2c is expressed in euryhaline species and how extrinsic and/or intrinsic factors modulate branchial clc-2c mRNA. Here, we investigated whether environmental salinity, prolactin (Prl) and osmotic conditions modulate clc-2c expression in euryhaline Mozambique tilapia (Oreochromis mossambicus). Branchial clc-2c and ncc2 mRNAs were enhanced in tilapia transferred from seawater (SW) to FW, whereas both mRNAs were attenuated upon transfer from FW to SW. Next, we injected hypophysectomized tilapia with ovine prolactin (oPrl) and observed a marked increase in clc-2c from saline-injected controls. To determine whether Prl regulates clc-2c in a gill-autonomous fashion, we incubated gill filaments in the presence of homologous tilapia Prls (tPrl177 and tPrl188). By 24 h, tPrl188 stimulated clc-2c expression ~5-fold from controls. Finally, filaments incubated in media ranging from 280 to 450 mosmol/kg for 3 and 6 h revealed that extracellular osmolality exerts a local effect on clc-2c expression; clc-2c was diminished by hyperosmotic conditions (450 mosmol/kg) compared with isosmotic controls (330 mosmol/kg). Our collective results suggest that hormonal and osmotic control of branchial clc-2c contributes to the FW adaptability of Mozambique tilapia. Moreover, we identify for the first time a regulatory link between Prl and a Clc Cl- channel in a vertebrate.

Hormonal regulation of aquaporin 3: opposing actions of prolactin and cortisol in tilapia gill.

Aquaporins (Aqps) are expressed within key osmoregulatory tissues where they mediate the movement of water and selected solutes across cell membranes. We leveraged the functional plasticity of Mozambique tilapia (Oreochromis mossambicus) gill epithelium to examine how Aqp3, an aquaglyceroporin, is regulated in response to osmoregulatory demands. Particular attention was paid to the actions of critical osmoregulatory hormones, namely, prolactin (Prl), growth hormone and cortisol. Branchial aqp3 mRNA levels were modulated following changes in environmental salinity, with enhanced aqp3 mRNA expression upon transfer from seawater to freshwater (FW). Accordingly, extensive Aqp3 immunoreactivity was localized to cell membranes of branchial epithelium in FW-acclimated animals. Upon transferring hypophysectomized tilapia to FW, we identified that a pituitary factor(s) is required for Aqp3 expression in FW. Replacement with ovine Prl (oPrl) was sufficient to stimulate Aqp3 expression in hypophysectomized animals held in FW, an effect blocked by coinjection with cortisol. Both oPrl and native tilapia Prls (tPrl177 and tPrl188) stimulated aqp3 in incubated gill filaments in a concentration-related manner. Consistent with in vivo responses, coincubation with cortisol blocked oPrl-stimulated aqp3 expression in vitro Our data indicate that Prl and cortisol act directly upon branchial epithelium to regulate Aqp3 in tilapia. Thus, within the context of the diverse actions of Prl on hydromineral balance in vertebrates, we define a new role for Prl as a regulator of Aqp expression.