An anion exchanger mediates bile acid transport across the placental microvillous membrane.
Microvillous membrane vesicles from the term human placental syncytiotrophoblast were used to characterize further the properties of a transport mechanism for bile acids. Taurocholate (TC) uptake into an osmotically reactive intravesicular space was temperature dependent and independent of sodium. TC uptake (2 microM) was markedly inhibited by 250 microM taurine and glycine-conjugated cholate and chenodeoxycholate and unconjugated cholate but not by chenodeoxycholate, deoxycholate, etianic acid, bromosulfophthalein, pyruvate, lactate, alanine, or taurine. The initial rate of TC uptake was inhibited significantly by the anion transport inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) but was not inhibited significantly by 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid, amiloride, or furosemide. Preincubation of vesicles with DIDS in the presence of TC partially blocked the action of the inhibitor. Efflux of 5 microM TC from membrane vesicles was stimulated by the presence of 50 microM TC in the incubation media. Basal as well as transstimulated TC efflux was inhibited by DIDS. The initial rate of TC influx followed saturation kinetics with an apparent Michaelis constant of 112 +/- 23 microM and maximal velocity of 2.01 +/- 0.19 nmol.mg protein-1.min-1. When the transmembrane electrical potential difference across the brush-border membrane vesicles was altered by external anion replacement or by valinomycin-induced K+ diffusion potentials, TC uptake was not significantly affected. DIDS-sensitive TC uptake was stimulated two-to threefold by an outwardly directed hydroxyl gradient (pH 7.7in/5.5out) compared with TC influx under pH-equilibrated conditions (pH 7.7in/7.7out). These studies are consistent with an electroneutral anion-exchange mechanism that mediates transfer of conjugated bile acids across the microvillous membrane of the syncytiotrophoblast.
Dumaswala
R
R
Division of Pediatric Gastroenterology, Children's Hospital Medical Center, Cincinnati, Ohio 45229.
Setchell
K D
KD
Moyer
M S
MS
Suchy
F J
FJ
D000837:Anion Exchange Resins / Q000378:metabolism*; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006913:Hydroxysteroid Dehydrogenases; D007477:Ions; D008562:Membrane Glycoproteins; D008564:Membrane Potentials; D008871:Microvilli / Q000378:metabolism / Q000502:physiology; D010920:Placenta / Q000378:metabolism* / Q000502:physiology; D011247:Pregnancy; D013379:Substrate Specificity; D013656:Taurocholic Acid / Q000493:pharmacokinetics*; D013696:Temperature; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)
D000837:Anion Exchange Resins; D002352:Carrier Proteins; D006913:Hydroxysteroid Dehydrogenases; D007477:Ions; D008562:Membrane Glycoproteins; D013656:Taurocholic Acid; C037643:bile acid binding proteins; C435496:AKR1C2 protein, human; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)