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  • 1
    ISSN: 1432-1424
    Keywords: rabbit ileum ; sulfate influx ; sodium dependence ; pH dependence ; NaSO 4 − ion pair ; lithium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary In intact ileal mucosa, uptake of SO4 across the brush border membrane requires the presence of Na and is saturable, withK1/2=1.3mm at 140mm Na (P.L. Smith, S.A. Orellana & M. Field, 1981.J. Membrane Biol. 63:199–206). The present study examines the substrate specificities and transport stoichiometry of the Na-dependent SO4 uptake process. The effects of variations in medium anion and cation composition on lumen-to-epithelium influx of SO4 (J me SO4 ) were determined under short-circuit conditions.J me SO4 is inhibited by thiosulfate, but not by phosphate, methylsulfate, vanadate or taurocholate. Cl is weakly inhibitory. Uptake of SO4 is poorly supported by Li, and is unaffected by K, indicating a specific dependence on Na. At low SO4 concentration (0.22mm),J me SO4 is a hyperbolic function of medium Na concentration; the corresponding Hill plot is linear with a slope of 1.0, suggesting a transport stoichiometry of 1 Na: 1 SO4. At high SO4 concentration (6.7mm), the Na-dependent SO4 velocity curve is sigmoidal and yields a Hill plot which is again linear but has a slope of 1.56, suggesting transport of more than 1 Na per SO4. SO4 uptake in presence of Na exhibits a dependence on medium pH. At 0.22mm SO4 and 140mm Na,J me SO4 was doubled by lowering pH from 7.4 to 6.8. However, at 6.7mm SO4 and 140mm Na, changing pH had no effect onJ me SO4 over the range 6.8 to 8.5. The pH dependence ofJ me SO4 at 6.7mm SO4 was restored when medium Na was lowered to 3mm, suggesting that pH sensitivity is a function of the concentration of preformed NaSO 4 − ion pair. The results suggest that SO4 influx across the ileal brush border occurs by electroneutral Na+/NaSO 4 − or Na+/H+/SO 4 2− cotransport, the former being favored by high concentrations of Na and SO4.
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  • 2
    ISSN: 1432-1424
    Keywords: rabbit ileum ; sulfate transport ; sodium dependence ; chloride dependence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Unidirectional fluxes of35SO4 across and into rabbit ileal epithelium were measured under short-circuit conditions, mostly at a medium SO4 concentration of 2.4mm. Unidirectional mucosa (m)-to-serosa (s) ands-to-m fluxes (J ms,J sm) were 0.456 and 0.067 μmoles hr−1 cm−2, respectively.J ms was 2.7 times higher in distal ileum than in mid-jejunum. Ouabain abolished net SO4 transport (J net) by reducingJ ms. Epinephrine, a stimulus of Cl absorption, had no effect on SO4 fluxes. Theophylline, a stimulus of Cl secretion, reducedJ ms without affectingJ sm, causing a 33% reduction inJ net. Other secretory stimuli (8-Br-cAMP, heat-stable enterotoxin, Ca-ionophore A23187) had similar effects. Replacement of all Cl with gluconate markedly reducedJ net through both a decrease inJ ms and an increase inJ sm. The anion-exchange inhibitor, 4-acetoamido-4′-isothiocyano-2,2′-sulfonic acid stilbene (SITS), when added to the serosal side, reducedJ ms by 94%, nearly abolishingJ net. SITS also decreasedJ sm by 75%. Mucosal SITS (50 μm) was ineffective. 4,4′-diisothiocyano-2,2′-sulfonic acid stilbene (DIDS) had effects similar to SITS but was less potent. Measurements of initial rates of epithelial uptake from the luminal side (J me) revealed the following: (1)J me is a saturable function of medium concentration with aV max of 0.94 μmoles hr−1 cm−2 and aK 1/2 of 1.3mm; (2) replacing all Na with choline abolishedJ me; (3) replacing all Cl with gluconate increasedJ me by 40%; (4) serosal SITS had no effect onJ me; and (5) stimuli of Cl secretion had no effect onJ me or increased it slightly. Determination of cell SO4 with35SO4 indicated that, at steady-state, the average mucosal concentration is 1.1 mmoles per liter cell water, less than half the medium concentration. Cell SO4 was increased to 3.0mm by adding SITS to the serosal side. Despite net transport rates greater than 1.4 μEq hr−1 cm−2, neither addition of SO4 to the SO4-free medium nor addition of SITS to SO4-containing medium altered short-circuit current. The results suggest that (1) ileal SO4 absorption consists of Na-coupled influx (symport) across the brush border and Cl-coupled efflux (antiport) across the basolateral membrane; (2) the overall process is electrically neutral; (3) the medium-to-cell Cl concentration difference may provide part of the driving force for net SO4 absorption; and (4) since agents affecting Cl fluxes (both absorptive and secretory) have little effect on SO4 fluxes, the mechanisms for their transcellular transports are under separate regulation.
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