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Am J Physiol Gastrointest Liver Physiol 283: G1004-G1013, 2002; doi:10.1152/ajpgi.00158.2002
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Vol. 283, Issue 4, G1004-G1013, October 2002

ClC-2 in guinea pig colon: mRNA, immunolabeling, and functional evidence for surface epithelium localization

Marcelo Catalán1, Isabel Cornejo1, Carlos D. Figueroa2, María Isabel Niemeyer1, Francisco V. Sepúlveda1, and L. Pablo Cid1

1 Centro de Estudios Científicos, Valdivia, and 2 Instituto de Histología y Patología, Universidad Austral de Chile, Valdivia, Chile

The principal function of the colon in fluid homeostasis is the absorption of NaCl and water. Apical membrane Na+ channels, Na+/H+ and Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchangers, have all been postulated to mediate NaCl entry into colonocytes. The identity of the basolateral exit pathway for Cl- is unknown. We have previously demonstrated the presence of the ClC-2 transcript in the guinea pig intestine. Now we explore in more detail, the tissue and cellular distribution of chloride channel ClC-2 in the distal colon by in situ hybridization and immunohistochemistry. The patch-clamp technique was used to characterize Cl- currents in isolated surface epithelial cells from guinea pig distal colon and these were compared with those mediated by recombinant guinea pig (gp)ClC-2. ClC-2 mRNA and protein were found in the surface epithelium of the distal colon. Immunolocalization revealed that, in addition to some intracellular labeling, ClC-2 was present in the basolateral membranes but absent from the apical pole of colonocytes. Isolated surface epithelial cells exhibited hyperpolarization-activated chloride currents showing a Cl- > I- permeability and Cd2+ sensitivity. These characteristics, as well as some details of the kinetics of activation and deactivation, were very similar to those of recombinant gpClC-2 measured in parallel experiments. The presence of active ClC-2 type currents in surface colonic epithelium, coupled to a basolateral location for ClC-2 in the distal colon, suggests a role for ClC-2 channel in mediating basolateral membrane exit of Cl- as an essential step in a NaCl absorption process.

hyperpolarization-activated chloride currents; in situ hybridization; immunohistochemistry


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