title: Electrogenic bicarbonate secretion by prairie dog gallbladder.
abstract: Pathological rates of gallbladder salt and water transport may promote the formation of cholesterol gallstones. Because prairie dogs are widely used as a model of this event, we characterized gallbladder ion transport in animals fed control chow by using electrophysiology, ion substitution, pharmacology, isotopic fluxes, impedance analysis, and molecular biology. In contrast to the electroneutral properties of rabbit and Necturus gallbladders, prairie dog gallbladders generated significant short-circuit current (I(sc); 171 +/- 21 microA/cm(2)) and lumen-negative potential difference (-10.1 +/- 1.2 mV) under basal conditions. Unidirectional radioisotopic fluxes demonstrated electroneutral NaCl absorption, whereas the residual net ion flux corresponded to I(sc). In response to 2 microM forskolin, I(sc) exceeded 270 microA/cm(2), and impedance estimates of the apical membrane resistance decreased from 200 Omega.cm(2) to 13 Omega.cm(2). The forskolin-induced I(sc) was dependent on extracellular HCO(3)(-) and was blocked by serosal 4,4'-dinitrostilben-2,2'-disulfonic acid (DNDS) and acetazolamide, whereas serosal bumetanide and Cl(-) ion substitution had little effect. Serosal trans-6-cyano-4-(N-ethylsulfonyl-N-methylamino)-3-hydroxy-2,2-dimethyl-chroman and Ba(2+) reduced I(sc), consistent with the inhibition of cAMP-dependent K(+) channels. Immunoprecipitation and confocal microscopy localized cystic fibrosis transmembrane conductance regulator protein (CFTR) to the apical membrane and subapical vesicles. Consistent with serosal DNDS sensitivity, pancreatic sodium-bicarbonate cotransporter protein pNBC1 expression was localized to the basolateral membrane. We conclude that prairie dog gallbladders secrete bicarbonate through cAMP-dependent apical CFTR anion channels. Basolateral HCO(3)(-) entry is mediated by DNDS-sensitive pNBC1, and the driving force for apical anion secretion is provided by K(+) channel activation.
authors.affiliation: Univ. of Pittsburgh School of Medicine, 497 Scaife Hall; 3550 Terrace St.; Pittsburgh, PA 15261, USA. moseraj@upmc.edu
authors.forename: A James
authors.initials: AJ
authors.lastname: Moser
authors.forename: A
authors.initials: A
authors.lastname: Gangopadhyay
authors.forename: N A
authors.initials: NA
authors.lastname: Bradbury
authors.forename: K W
authors.initials: KW
authors.lastname: Peters
authors.forename: R A
authors.initials: RA
authors.lastname: Frizzell
authors.forename: R J
authors.initials: RJ
authors.lastname: Bridges
mesh_terms: D000086:Acetazolamide / Q000494:pharmacology; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001639:Bicarbonates / Q000378:metabolism*; D002257:Carbonic Anhydrase Inhibitors / Q000494:pharmacology; D002712:Chlorides / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism*; D017097:Electric Impedance; D020536:Enzyme Activators / Q000494:pharmacology; D005704:Gallbladder / Q000187:drug effects / Q000378:metabolism*; D007213:Indomethacin / Q000494:pharmacology; D008297:Male; D008564:Membrane Potentials; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000378:metabolism*; D012589:Sciuridae; D012964:Sodium / Q000378:metabolism; D027982:Sodium-Bicarbonate Symporters / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013267:Stilbenes / Q000494:pharmacology
chemical_list: D001639:Bicarbonates; D002257:Carbonic Anhydrase Inhibitors; D002712:Chlorides; D016861:Cyclooxygenase Inhibitors; D020536:Enzyme Activators; D015221:Potassium Channels; D027982:Sodium-Bicarbonate Symporters; D013267:Stilbenes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005576:Colforsin; C022306:4,4'-dinitro-2,2'-stilbenedisulfonic acid; D012964:Sodium; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D000086:Acetazolamide; D011188:Potassium; D007213:Indomethacin