Na absorption across endometrial epithelial cells is stimulated by cAMP-dependent activation of an inwardly rectifying K channel.
Previous studies in our laboratory have shown that Na absorption across the porcine endometrium is stimulated by PGF2alpha and cAMP-dependent activation of a barium-sensitive K channel located in the basolateral membrane of surface epithelial cells. In this study, we identify and characterize this basolateral, barium-sensitive K conductance. Porcine uterine tissues were mounted in Ussing chambers and bathed with KMeSO4 Ringer solution. Amphotericin B (70 microM) was added to the luminal solution to permeabilize the apical membrane and determine the current-voltage relationship of the basolateral K conductance after activation by 100 microM CPT-cAMP. An inwardly rectifying current was identified which possessed a reversal potential of -53 mV when standard Ringer solution was used to bathe the serosal surface. The K:Na selectivity ratio was calculated to be 12:1. Administration of 5 mM barium to the serosal solution completely inhibited the current activated by cAMP under these conditions. In addition to these experiments, amphotericin-perforated whole cell patch clamp recordings were obtained from primary cultures of porcine surface endometrial cells. The isolated cells displayed an inwardly rectifying current under basal conditions. This current was significantly stimulated by CPT-cAMP and blocked by barium. These results together with our previous studies demonstrate that cAMP increases Na absorption in porcine endometrial epithelial cells by activating an inwardly rectifying K channel present in the basolateral membrane. Similar patch clamp experiments were conducted using cells from a human endometrial epithelial cell line, RL95-2. An inwardly rectifying current was also identified in these cells which possessed a reversal potential of -56 mV when the cells were bathed in standard Ringer solution. This current was blocked by barium as well as cesium. However, the current from the human cells did not appear to be activated by cAMP, indicating that distinct subtypes of inwardly rectifying K channels are present in endometrial epithelial cells from different species.
Vetter
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Department of Veterinary PathoBiology, 295 Animal Science/Veterinary Medicine Building, University of Minnesota, St. Paul, Minnesota 55108, USA.
Deachapunya
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C
O'Grady
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SM
D000042:Absorption; D000818:Animals; D001464:Barium / Q000494:pharmacology; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D004717:Endometrium / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D017136:Ion Transport / Q000187:drug effects; D008564:Membrane Potentials; D008954:Models, Biological; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism*; D013045:Species Specificity; D013552:Swine; D013873:Thionucleotides / Q000494:pharmacology
D015221:Potassium Channels; D013873:Thionucleotides; D001464:Barium; C015929:8-((4-chlorophenyl)thio)cyclic-3',5'-AMP; D012964:Sodium; D000242:Cyclic AMP