Functional activity of Pat-1 (Slc26a6) Cl(−)/HCO₃(−) exchange in the lower villus epithelium of murine duodenum

Abstract

Aims: The apical membrane anion exchanger putative anion transporter-1 (Pat-1) is expressed at significant levels in the lower villus epithelium of murine duodenum. However, previous studies of Cl(−)/HCO₃(−) exchange in the lower villus have failed to demonstrate Pat-1 function. Those studies routinely included luminal glucose which induces Na(+) -coupled glucose transport and acidifies the villus epithelium. Since Pat-1 has been proposed to be an electrogenic 1Cl(−)/2HCO₃(−) exchanger, membrane depolarization or cell acidification during glucose transport may obscure Pat-1 activity. Therefore, we investigated the effects of luminal glucose on Cl(−)(IN)/HCO₃(−) (OUT) exchange activity in the lower villus epithelium.

Methods: Cl(−)(IN) /HCO (−) (OUT) exchange of villus epithelium in duodenal mucosa from Pat-1 knockout (KO), Slc26a3 [down-regulated in adenoma (Dra)] KO, cystic fibrosis transmembrane conductance regulator (Cftr) KO and wild-type (WT) littermate mice was measured using the pH-sensitive dye 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein. Short-circuit current (I(sc) ) was measured in Ussing chambers.

Results: During glucose absorption, Cl(−)(IN)/HCO₃(−) (OUT) exchange in the lower villus epithelium was abolished in the Dra KO and unaffected in the Pat-1 KO relative to WT. However, during electroneutral mannose absorption or electrogenic α-D-methyl glucoside absorption, Cl(−)(IN) /HCO₃(−) (OUT) exchange was reduced in both Pat-1 KO and Dra KO villi. Exposure to high [K(+)] abolished Cl(−)(IN) /HCO₃(−) (OUT) exchange in the Dra KO but not the Dra/Cftr double KO epithelium, suggesting that Pat-1 activity is little affected by membrane depolarization except in the presence of Cftr.

Conclusions: The metabolic and electrogenic activity of glucose transport obscures Cl(−)(IN) /HCO₃(−) (OUT) exchange activity of Pat-1 in the lower villus. The inhibitory effects of membrane depolarization on Pat-1 Cl(−)(IN) /HCO₃(−) (OUT) exchange may require concurrent membrane association with Cftr.