title: Autosomal dominant polycystic kidney disease decreases anion exchanger activity.
abstract: Liver cysts, the most common extrarenal manifestation of autosomal dominant polycystic kidney disease (ADPKD), derive from the intrahepatic biliary epithelium (IBE) and are found in 60-75% of ADPKD patients on dialysis. Secretin-induced secretion by the normal IBE is rich in HCO3-, whereas intact ADPKD liver cysts secrete primarily Cl- in response to secretin. To evaluate the mechanisms of decreased HCO3- secretion by ADPKD liver cysts, we utilized SV40 large T antigen-immortalized normal IBE and ADPKD liver cyst-derived epithelia (LCDE) cell lines that we created. These cell lines express biliary but not hepatocyte markers. Anion exchanger (AE) function was assessed by the response of intracellular pH (pHi) to acute Cl- removal. 2',7'-Bis(carboxyethyl)-5-(6)-carboxyfluorescein-loaded monolayers were continuously perfused with physiological HCO3- buffer containing Cl- or gluconate. In IBE cell line H75 (n = 6), acute Cl- removal alkalinized pHi at a rate of 0.04 +/- 0.01 min-1. AE function was significantly decreased in LCDE cell line CL3 (n = 6) to a rate of 0.01 +/- 0.01 min-1 after Cl- removal. Northern blot analysis demonstrated equivalent levels of AE2 mRNA in both cell lines. AE1 mRNA was undetectable. Immunoblot analysis demonstrated the AE2 polypeptide in both cell lines, but the level of mature glycosylated AE2 polypeptide was reduced in LCDE cells. Immunofluorescence microscopy demonstrated decreased membrane-localized AE2 in LCDE cells. These findings suggest that decreased plasmalemmal AE2 may account for decreased AE function in LCDE cells and suggest a possible explanation for decreased secretion of HCO3- by ADPKD liver cysts.
authors.affiliation: Department of Medicine, New England Medical Center, Boston, Massachusetts, USA.
authors.forename: R D
authors.initials: RD
authors.lastname: Perrone
authors.forename: S A
authors.initials: SA
authors.lastname: Grubman
authors.forename: S L
authors.initials: SL
authors.lastname: Murray
authors.forename: D W
authors.initials: DW
authors.lastname: Lee
authors.forename: S L
authors.initials: SL
authors.lastname: Alper
authors.forename: D M
authors.initials: DM
authors.lastname: Jefferson
mesh_terms: D000468:Alkalies / Q000494:pharmacology; D017920:Antiporters / Q000378:metabolism*; D001653:Bile Ducts, Intrahepatic / Q000166:cytology / Q000378:metabolism; D015153:Blotting, Western; D002021:Buffers; D002461:Cell Line, Transformed; D027963:Chloride-Bicarbonate Antiporters; D002712:Chlorides / Q000378:metabolism; D003560:Cysts / Q000378:metabolism / Q000473:pathology; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000378:metabolism; D008107:Liver Diseases / Q000378:metabolism / Q000473:pathology; D016891:Polycystic Kidney, Autosomal Dominant / Q000378:metabolism* / Q000473:pathology; D012016:Reference Values; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014018:Tissue Distribution
chemical_list: D000468:Alkalies; D017920:Antiporters; D002021:Buffers; D027963:Chloride-Bicarbonate Antiporters; D002712:Chlorides; D013755:Tetradecanoylphorbol Acetate