Fluid transport in a cultured cell model of kidney epithelial cyst enlargement

George A. Tanner, Michele R. Maxwell, James A. McAteer

Research output: Contribution to journalArticle

22 Scopus citations

Abstract

Madin-Darby canine kidney (MDCK) cells, when seeded into collagen gel, form polarized, spherical, epithelial cysts, which grow by a process involving fluid secretion and cell proliferation. These cysts are a useful model for understanding the dynamics of cyst enlargement in renal cystic disease. The hypothesis that MDCK cyst fluid secretion depends upon chloride secretion was tested, and a cell model for this process is presented here. Lumen and epithelial cell volumes were measured by video microscopy in acute experiments. Fluid absorption (-0.073 ± 0.007 μl·h-1·cm-2; N = 8) was observed when cysts were superfused with unsupplemented Dulbecco's modified Eagle's medium at 36 to 37°C. Fluid secretion (0.221 ± 0.016 μl·h-1·cm-2; N = 25) was seen when 1 mM dibutyryl cAMP plus 0.1 mM 3-isobutyl-1-methylxanthine were added to the superfusate. cAMP-induced fluid secretion was significantly inhibited by basolateral 1 mM ouabain, 0.1 mM furosemide, or 1 mM amiloride. It was not significantly affected by 1 mM chlorothiazide, 0.01 mM bumetanide, or 0.1 mM acetazolamide in the presence of normal bicarbonate/CO2. In the nominal absence of bicarbonate/CO2, fluid secretion was 18% of control. Vasopressin-induced fluid secretion was significantly inhibited by pretreatment of cysts with 0.1 mM 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS). Cyst cell shrinkage in isosmotic chloride-free Ringer's solution (chloride replaced by gluconate) was inhibited by 0.1 mM basolateral DIDS. The results suggest that chloride-bicarbonate exchange in the basolateral membrane of MDCK cyst epithelial cells plays a critical role in cyst fluid secretion.

Original languageEnglish (US)
Pages (from-to)1208-1218
Number of pages11
JournalJournal of the American Society of Nephrology
Volume2
Issue number7
StatePublished - Jan 1 1992

Keywords

  • cAMP
  • Chloride transport
  • Chloride/bicarbonate exchange
  • Madin-Darby canine kidney cells
  • Renal cystic disease

ASJC Scopus subject areas

  • Nephrology

Fingerprint Dive into the research topics of 'Fluid transport in a cultured cell model of kidney epithelial cyst enlargement'. Together they form a unique fingerprint.

  • Cite this