Ontogeny of bile acid transport in brush border membrane vesicles from rat ileum

Abstract

We studied the postnatal development of bile acid transport in rat ileum, using brush border membrane vesicles prepared by a Ca2+ precipitation method. Membrane vesicles from developing (day 14–21) and adult Sprague-Dawley rats were enriched to a similar degree in brush border membrane marker enzyme activities (sucrase or lactase) compared with homogenate. Uptake of 25 μM [3H]taurocholate by adult membrane vesicles was markedly accelerated in the presence of an inwardly directed 100 mM Na+ gradient compared with a K+ gradient, and there was a transient intravesicular accumulation of isotope above equilibrium (“overshoot”). In contrast, at 14 and 16 days of age there was no difference in taurocholate uptake in the presence of a Na+ or a K+ gradient, and uptake was not saturable. The integrity of the vesicle preparation from 14- and 16-day-old rats was confirmed by the demonstration of Na+-dependent uphill transport of 100 μM l-[3H]alanine. Stimulation of taurocholate uptake by a Na+ compared with a K+ gradient (“sodium effect”) was first observed at age 17 days, but an overshoot was not present until 18 days of age. The initial rate of Na+-dependent taurocholate (25 μM) uptake increased sixfold between 17 and 21 days of age (24.36 ± 6.11 to 148.59 ± 8.56 pmol · mg−1 protein · 5 s−1). Absent or decreased Na+-dependent taurocholate uptake was not due to increased permeability or “leakiness” of vesicles from younger animals to Na+. Ileal brush border membrane vesicles demonstrated saturable kinetics at 21 days, but the Vmax was significantly lower (10.15 ± 0.44 vs. 13.42 ± 0.59 nmol · mg−1 protein · min−1, p < 0.001) and the apparent Km higher (130.6 ± 18.9 vs. 70.1 ± 12.6 μM, p < 0.007) than the adult. We conclude that (a) saturable, Na+-bile acid coupled transport is absent in rat ileum throughout most of the suckling period and (b) kinetic analysis suggests that maturation occurs near weaning, primarily through an increase in functional bile acid carriers within the ileal brush border membrane.