Regulation of electrolyte transport across cultured endometrial epithelial cells by prolactin.
The effect of prolactin (PRL) on ion transport across the porcine glandular endometrial epithelial cells was studied in primary cell culture using the short-circuit current technique. Addition of 1 microg/ml PRL either to the apical solution or to the basolateral solution produced a peak followed by a sustained increase in Isc, but with a lesser response when PRL was added apically. Basolateral addition of PRL increased the Isc in a concentration-dependent manner with a maximum effect at 1 microg/ml and an effective concentration value of 120 ng/ml. The PRL-stimulated Isc was significantly reduced by pretreatment with an apical addition of 5-nitro-2-(3-phenylpropylamino) benzoic acid (200 microM), diphenylamine-2-carboxylic acid (1 mM) or 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (200 microM), Cl(-) channel blockers, but not by amiloride (10 microM), a Na(+) channel blocker. In addition, pretreatment with bumetanide (200 microM), a Na(+)-K(+)-2Cl(-) cotransporter inhibitor, in the basolateral solution significantly reduced the PRL-stimulated Isc. Replacement of Cl(-) or in the bathing solutions also decreased the Isc response to PRL. Pretreatment of the monolayer with AG490 (50 microM), an inhibitor of JAK2 activity significantly inhibited the PRL-induced increase in Isc. Western blot analysis of the porcine endometrial epithelial cells revealed the presence of short isoform of PRL receptor (PRLR-S) that could be regulated by 17beta-estradiol. The results of this investigation showed that PRL acutely stimulated anion secretion across the porcine endometrial epithelial cells possibly through PRLR-S present in both apical and basolateral membranes. The PRL response appeared to be mediated by the JAK2-dependent pathway.
Deachapunya
Chatsri
C
Department of Physiology, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Wattana, Bangkok 10110, Thailand. chatsri@swu.ac.th
Poonyachoti
Sutthasinee
S
Krishnamra
Nateetip
N
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000818:Animals; D001639:Bicarbonates / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D004573:Electrolytes / Q000378:metabolism*; D004717:Endometrium / Q000166:cytology / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D017136:Ion Transport / Q000187:drug effects; D053614:Janus Kinase 2 / Q000502:physiology; D009579:Nitrobenzoates / Q000494:pharmacology; D011188:Potassium / Q000378:metabolism; D011388:Prolactin / Q000494:pharmacology*; D011981:Receptors, Prolactin / Q000032:analysis; D015398:Signal Transduction; D012964:Sodium / Q000378:metabolism; D013552:Swine
D001639:Bicarbonates; D002712:Chlorides; D004573:Electrolytes; D009579:Nitrobenzoates; D011981:Receptors, Prolactin; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D011388:Prolactin; D012964:Sodium; D053614:Janus Kinase 2; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D011188:Potassium