title: Prolactin stimulates sodium and chloride ion channels in A6 renal epithelial cells.
abstract: Many hormonal pathways contribute to the regulation of renal epithelial sodium channel (ENaC) function, a key process for maintaining blood volume and controlling blood pressure. In the present study, we examined whether the peptide hormone prolactin (PRL) regulates ENaC function in renal epithelial cells (A6). Basolateral application of several different concentrations of PRL dramatically stimulated the transepithelial current in A6 cells, increasing both amiloride-sensitive (ENaC) and amiloride-insensitive currents. Using cell-attached patch clamp, we determined that PRL increased both the number (N) and open probability (Po) of ENaC present in the apical membrane. Inhibition of PKA with H-89 abolished the effect of PRL on amiloride-sensitive and insensitive transepithelial currents and eliminated the increase in ENaC NPo with PRL exposure. PRL also increased cAMP in A6 cells, consistent with signaling through the cAMP-dependent PKA pathway. We also identified that PRL induced activity of a 2-pS anion channel with outward rectification, electrophysiological properties consistent with ClC4 or ClC5. RT-PCR only detected ClC4, but not ClC5 transcripts. Here, we show for the first time that PRL activates sodium and chloride transport in renal epithelial cells via ENaC and ClC4.
authors.affiliation: Department of Physiology, Emory University School of Medicine, Atlanta, Georgia.
authors.forename: Megan M
authors.initials: MM
authors.lastname: Greenlee
authors.affiliation: Department of Physiology, Emory University School of Medicine, Atlanta, Georgia.
authors.forename: Jeremiah D
authors.initials: JD
authors.lastname: Mitzelfelt
authors.affiliation: Department of Physiology, Emory University School of Medicine, Atlanta, Georgia.
authors.forename: Billie Jeanne
authors.initials: BJ
authors.lastname: Duke
authors.affiliation: Department of Physiology, Emory University School of Medicine, Atlanta, Georgia.
authors.forename: Otor
authors.initials: O
authors.lastname: Al-Khalili
authors.affiliation: Department of Physiology, Emory University School of Medicine, Atlanta, Georgia.
authors.forename: Hui-Fang
authors.initials: HF
authors.lastname: Bao
authors.affiliation: Department of Physiology, Emory University School of Medicine, Atlanta, Georgia deaton@emory.edu.
authors.forename: Douglas C
authors.initials: DC
authors.lastname: Eaton
mesh_terms: D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D018118:Chloride Channels / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D053503:Epithelial Sodium Channels / Q000378:metabolism*; D051379:Mice; D018408:Patch-Clamp Techniques / Q000379:methods; D011388:Prolactin / Q000494:pharmacology*; D012964:Sodium / Q000378:metabolism*
chemical_list: D018118:Chloride Channels; D053503:Epithelial Sodium Channels; D000584:Amiloride; D011388:Prolactin; D012964:Sodium; D000242:Cyclic AMP
keywords: ClC channels; ENaC; cAMP; hormones; osmoregulation; protein kinase A