Transfected rat cMOAT is functionally expressed on the apical membrane in Madin-Darby canine kidney (MDCK) cells

Pharm Res. 1998 Dec;15(12):1851-6. doi: 10.1023/a:1011953906065.


Purpose: The purpose of the present study is to investigate the expression of canalicular multispecific organic anion transporter (cMOAT) by its cDNA transfection in polarized Madin-Darby canine kidney cells (MDCK).

Methods: MDCK cells were transfected with an expression vector (pCXN2) containing the rat cMOAT cDNA with lipofectamine to obtain the stable transfectant under G418. Cells from a single colony whose cMOAT expression was the highest were seeded to form a tight epithelial monolayer on microporous membrane filters. Export of glutathione S-bimane (GS-B) from monolayers was determined after preloading its precursor, monochloro bimane (MCB).

Results: A comparable amount of GS-B was excreted to the apical and basal compartments in the vector-transfected cells. In contrast, in cMOAT-transfected cells, the amount apically excreted was approximately twice that excreted into the basal compartment. Cyclosporin A (CsA) (30 microM), an inhibitor of cMOAT at higher concentrations, inhibited the preferential apical export of GS-B from cMOAT-transfected cells.

Conclusions: Rat cMOAT is functionally expressed on the apical membrane of MDCK cells after transfection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anion Transport Proteins
  • Anions / metabolism
  • Biological Transport / drug effects
  • Biological Transport / physiology
  • Blotting, Northern
  • Bridged Bicyclo Compounds / pharmacokinetics
  • Carrier Proteins / genetics*
  • Carrier Proteins / metabolism*
  • Cation Exchange Resins
  • Cell Membrane / chemistry
  • Cell Membrane / metabolism
  • Cell Polarity / drug effects
  • Cyclosporine / pharmacology
  • DNA, Complementary
  • Dogs
  • Epithelial Cells / chemistry*
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism*
  • Fluorescent Dyes / pharmacokinetics
  • Gene Expression
  • Glutathione / analogs & derivatives
  • Glutathione / pharmacokinetics
  • Indicators and Reagents
  • Kidney / cytology
  • LLC-PK1 Cells
  • Lipids
  • Membrane Proteins / genetics
  • RNA, Messenger / analysis
  • Rats
  • Swine
  • Transfection


  • Anion Transport Proteins
  • Anions
  • Bridged Bicyclo Compounds
  • Carrier Proteins
  • Cation Exchange Resins
  • DNA, Complementary
  • Fluorescent Dyes
  • Indicators and Reagents
  • Lipids
  • Lipofectamine
  • Membrane Proteins
  • RNA, Messenger
  • glutathione-bimane
  • Cyclosporine
  • Glutathione