We have investigated the hypothesis that active
anion transport drives fluid secretion by the cystic epithelium in
autosomal dominant polycystic kidney disease (
ADPKD). We prepared monolayers of a primary culture derived from cystic tissue removed from
ADPKD patients. The monolayers were grown on permeant supports, and fluid secretion was initiated by
forskolin. The results were compared with those obtained with monolayers of Madin-Darby canine kidney (MDCK) cells, known to secrete Cl-. In the absence of the agonist,
ADPKD monolayers absorbed fluid (0.20 +/- 0.02 microliter.cm surface area-2.h-1).
Forskolin reversed this to secretion (0.60 +/- 0.03 microliter.cm-2.h-1). Control MDCK monolayers did not transport fluid in either direction, but
forskolin induced secretion (0.48 +/- 0.03 microliter.cm-2.h-1). The electrical properties of the monolayers were monitored in Ussing chambers.
Forskolin increased the transepithelial potential difference (Vte) of
ADPKD monolayers (-0.9 +/- 0.1 to -1.1 +/- 0.1 mV) and the short-circuit current (Isc) (6.6 +/- 0.7 to 9.2 +/- 0.8 microA/cm2). The transepithelial resistance (Rte) fell (156 +/- 9 to 138 +/- 10 omega.cm2). Similar results were obtained with MDCK monolayers. The polarity of Vte and the direction of the Isc are compatible with the hypothesis that active secretion of
anion drives fluid secretion. Basolateral application of the Na-K-2Cl cotransporter,
bumetanide, reduced
forskolin-stimulated fluid secretion by
ADPKD monolayers (0.56 +/- 0.05 to 0.28 +/- 0.03), depolarized Vte, and inhibited Isc without affecting Rte. Apical application of the Cl- channel blocker,
diphenylamine-2-carboxylate, also inhibited fluid secretion by
ADPKD monolayers (0.65 +/- 0.03 to 0.27 +/- 0.02 microliter.cm-2.h-1).(ABSTRACT TRUNCATED AT 250 WORDS)