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Updated: Jul 11, 2026

Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D
Published on: March 16, 2017
H+ and HCO3- flux across apical surface of rat distal colon
1Department of Medicine, Veterans Affairs Medical Center, Philadelphia, Pennsylvania.
Abstract:
Colonic ion transport is postulated to occur via simultaneous operation of Na(+)-H+ exchange and Cl(-)-HCO3- exchange. Accordingly H+ and HCO3- should be transported simultaneously by the colon. To assess simultaneous H+ and HCO3- transport, net acid-base flux was measured in isolated segments of rat distal colon. When both tissue surfaces were bathed in symmetrical solutions containing Cl-, net base was secreted (-1.0 +/- 0.1 mu eq.cm-2.h-1). Cl- substitution with gluconate in the mucosal medium caused net base flux to switch from secretion to absorption (2.0 +/- 0.2 mu eq.cm-2.h-1). To evaluate whether base absorption was dependent on H+ secretion via Na(+)-H+ exchange, mucosal Na+ was substituted with N-methylglucamine, and amiloride, an inhibitor of Na(+)-H+ exchange, was applied. Na+ substitution and 1 mM amiloride inhibited base absorption by 37 and 38%, respectively, suggesting operation of Na(+)-H+ exchange. Because base absorption persisted, an additional mechanism was considered, HCO3- absorption via Cl(-)-HCO3- exchange. This was evaluated with an inhibitor of Cl(-)-HCO3- exchange 4-acetamido-4'-isothiostilbene-2,2'-disulfonic acid (SITS). SITS (1 mM) inhibited HCO3- absorption by 40%. The effects of amiloride and SITS were additive, suggesting that the Na(+)-H+ and Cl(-)-HCO3- exchangers operate simultaneously. Amiloride also inhibited H+ secretion when net HCO3- was secreted, suggesting that the direction of HCO3- movement does not influence Na(+)-H+ exchange activity. These data suggest that the colon transports both H+ and HCO3- across the apical surface via Na(+)-H+ exchange and Cl(-)-HCO3- exchange; H+ is secreted via Na(+)-H+ exchange, whereas HCO3- can be secreted or absorbed via Cl(-)-HCO3- exchange.
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