Chapter 11. Transepithelial Glucose Transport

Introduction

Imagine that you are investigating the transepithelial transport of glucose. Intestinal epithelial cells are grown in culture to form a complete sheet so that the fluid bathing the apical region of the cells (the apical medium) is completely separated from the fluid bathing the basolateral region of the cells (the basolateral medium). Radioactive (¹⁴C-labeled) glucose is added to the apical medium, and the appearance of radioactivity in the basolateral medium is monitored in terms of counts per minute per milliliter (cpm/ml), a measure of radioactivity per unit volume.

Treatment 1: The apical and basolateral media each contain 150 mM Na⁺ (curve 1).

Treatment 2: The apical medium contains 1 mM Na⁺, and the basolateral medium contains 150 mM Na⁺ (curve 2).

Treatment 3: The apical medium contains 150 mM Na⁺, and the basolateral medium contains 1 mM Na⁺ (curve 3).

Question

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Transepithelial transport of glucose requires the cotransport of Na⁺ down its concentration gradient to drive glucose against its concentration gradient from the apical medium into the cells. This coupling cannot occur when the concentration of Na⁺ in the apical medium is low, as is the case for curve 2. Under these conditions, there would be no inward gradient of Na⁺ across the apical membrane to drive glucose uptake. Subsequently, for the glucose to move from the cells into the basolateral medium, the glucose is transported down its concentration gradient and thus does not depend on Na⁺ and its concentration in the basolateral medium. Accordingly, the concentration of Na⁺ in the basolateral medium is not a factor.

Question

In additional studies, the drug ouabain, which inhibits Na⁺/K⁺ ATPases, is included as noted.

Treatment 4: The apical and basolateral media contain 150 mM Na⁺ and the apical medium contains ouabain (curve 4).

Treatment 5: The apical and basolateral media contain 150 mM Na⁺ and the basolateral medium contains ouabain (curve 5).

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_feedback: In order for the epithelial cells to maintain a low Na⁺ concentration, the Na⁺ must be continually pumped out of the cells by the Na⁺/K⁺ ATPase. Ouabain inhibits this pump and thus would prevent the cells from keeping their cytosolic [Na⁺] low. If Na⁺ cannot be pumped out, then the Na⁺ gradient driving glucose transport into the cells would dissipate, and glucose transport into the cells would cease. The observation that glucose transport ceases only when ouabain is added to the basolateral medium suggests that the Na⁺/K⁺ ATPase is localized at and/or only functions in this membrane domain.

Question

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_feedback: The drug inhibits the Na⁺-glucose symporter.