Molecular targets of prolactin in mummichogs (Fundulus heteroclitus): Ion transporters/channels, aquaporins, and claudins.

Prolactin (Prl) was identified over 60 years ago in mummichogs (Fundulus heteroclitus) as a "freshwater (FW)-adapting hormone", yet the cellular and molecular targets of Prl in this model teleost have remained unknown. Here, we conducted a phylogenetic analysis of two mummichog Prl receptors (Prlrs), designated Prlra and Prlrb, prior to describing the tissue- and salinity-dependent expression of their associated mRNAs. We then administered ovine Prl (oPrl) to mummichogs held in brackish water and characterized the expression of genes associated with FW- and seawater (SW)-type ionocytes. Within FW-type ionocytes, oPrl stimulated the expression of Na/Cl cotransporter 2 (ncc2) and aquaporin 3 (aqp3). Alternatively, branchial Na/H exchanger 2 and -3 (nhe2 and -3) expression did not respond to oPrl. Gene transcripts associated with SW-type ionocytes, including Na/K/2Cl cotransporter 1 (nkcc1), cystic fibrosis transmembrane regulator 1 (cftr1), and claudin 10f (cldn10f) were reduced by oPrl. Isolated gill filaments incubated with oPrl in vitro exhibited elevated ncc2 and prlra expression. Given the role of Aqps in supporting gastrointestinal fluid absorption, we assessed whether several intestinal aqp transcripts were responsive to oPrl and found that aqp1a and -8 levels were reduced by oPrl. Our collective data indicate that Prl promotes FW-acclimation in mummichogs by orchestrating the expression of solute transporters/channels, water channels, and tight-junction proteins across multiple osmoregulatory organs.