H+/solute-induced intracellular acidification leads to selective activation of apical Na+/H+ exchange in human intestinal epithelial cells

The intestinal absorption of many nutrients and. drug molecules is mediated by ion-driven transport mechanisms in the intestinal enterocyte plasma mem brane. Clearly, the establishment and maintenance of the driving forces - t ransepithelial ion gradients - are vital for maximum nutrient absorption. T he purpose of this study was to determine the nature of intracellular intra cellular pH (pH(i)) regulation in response to H+-coupled transport at the a pical membrane of human intestinal epithelial Caco-2 cells. Using isoform-s pecific primers, mRNA transcripts of the Na+/H+ exchangers NHE1, NHE2, and NHE3 were detected by RT-PCR, and identities were confirmed by sequencing. The functional profile of Na+/H+ exchange was determined by a combination o f pHi, Na-22(+) influx, and EIPA inhibition experiments. Functional NHE1 an d NHE3 activities were identified at the basolateral and apical membranes, respectively. H+/solute-induced acidification (using glycylsarcosine or bet a-alanine) led to Na+-dependent, EIPA-inhibitable pH(i) recovery or EIPA-in hibitable Na-22(+) influx at the apical membrane only. Selective activation of apical (but not basolateral) Na+/H+ exchange by H+/solute cotransport d emonstrates that coordinated activity of H+/solute symport with apical Na+/ H+ exchange optimizes the efficient absorption of nutrients and Na+, while maintaining pH(i) and the ion gradients involved in driving transport.