Expression, Localization and Functional Activity of the Major Na⁺/H⁺ Exchange Isoforms Expressed in the Intestinal Cell Line Caco-2BBe.

ABSTRACT:

Background/aims

Enterocytes express a number of NHE isoforms with presumed localization in the apical (NHE2, 3 and 8) or basolateral (NHE1) membrane. Functional activity and localization of enterocyte NHE isoforms were assessed using fully differentiated Caco-2BBe cells, whose genetic expression profile closely resembles mature enterocytes.

Methods

The activity of the different NHEs was analyzed by fluorometric pH_i-metry in a perfusion chamber with separate apical and basolateral perfusion, using specific inhibitors and shRNA knockdown of NHE2. The expression of the NHEs and of other relevant acid extrusion transporters was quantified by qPCR.

Results

Quantitative comparison of the mRNA expression levels of the different NHE isoforms in 14 day-differentiate

SUBMITTER: Yu Y 

PROVIDER: S-EPMC7104346 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature