Na-H exchangers prevent cellular acidification by catalyzing the electroneutral exchange of extracellular sodium for an intracellular proton. To date, seven Na-H exchangers have been identified in mammals, and although several members of this family have been extensively studied and characterized, it is clear that there are major gaps in our understanding with respect to the remaining family members. In order to initiate the study of Na-H exchangers in a genomically-defined and genetically-tractable model system, we have cloned the complete cDNAs and analyzed splice site variation for nine putative homologs from the nematode C. elegans, which we have called nhx-1
through -9. The expression patterns and cellular distributions of the nhx proteins were determined using transcriptional and translational promoter-transgene fusion constructs to Green Fluorescent Protein (GFP). Four of the putative exchangers were expressed at the cell surface, while five of the exchangers were associated with the membranes of intracellular organelles. Individual isoforms were expressed exclusively in the intestine, seam cells, hypodermal cells of the main body syncytium, and the excretory cell, all of which are polarized epithelial cells, suggesting a role for these proteins in epithelial membrane transport processes in the nematode. Other isoforms were found to express either ubiquitously, or in a pan-neural pattern, suggesting a more conserved role in cell pH regulation or neuronal function. Finally, we show that recombinant nhx-4
, the ubiquitous nematode Na-H exchanger, mediates Na+-dependent pH recovery following intracellular acidification. Nhx-4 activity has a Ka for Na+ of approximately 32 mM, is not Cl--dependent, and is relatively insensitive to the amiloride analog EIPA.