Na⁺/H⁺ exchanger isoform 1 activity in AQP2-expressing cells can be either proliferative or anti-proliferative...

We have previously shown in renal cells that expression of the water channel Aquaporin-2 increases cell proliferation by a regulatory volume mechanism involving Na⁺/H⁺ exchanger isoform 2. Here, we investigated if Aquaporin-2 (AQP2) also modulates Na⁺/H⁺ exchanger isoform 1-dependent cell proliferation. We use two AQP2-expressing cortical collecting duct models: one constitutive (WT or AQP2-transfected RCCD₁ cell line) and one inducible (control or vasopressin-induced mpkCCD_c14 cell line). We found that Aquaporin-2 modifies Na⁺/H⁺ exchanger isoform 1 (NHE1) contribution to cell proliferation. In Aquaporin-2-expressing cells, Na⁺/H⁺ exchanger isoform 1 is anti-proliferative at physiological pH. In acid media, Na⁺/H⁺ exchanger isoform 1 contribution turned from anti-proliferative to proliferative only in AQP2-expressing cells. We also found that, in AQP2-expressing cells, NHE1-dependent proliferation changes parallel changes in stress fiber levels: at pH 7.4, Na⁺/H⁺ exchanger isoform 1 would favor stress fiber disassembly and, under acidosis, NHE1 would favor stress fiber assembly. Moreover, we found that Na⁺/H⁺ exchanger-dependent effects on proliferation linked to Aquaporin-2 relied on Transient Receptor Potential Subfamily V calcium channel activity. In conclusion, our data show that, in collecting duct cells, the water channel Aquaporin-2 modulates NHE1-dependent cell proliferation. In AQP2-expressing cells, at physiological pH, the Na⁺/H⁺ exchanger isoform 1 function is anti-proliferative and, at acidic pH, Na⁺/H⁺ exchanger isoform 1 function is proliferative. We propose that Na⁺/H⁺ exchanger isoform 1 modulates proliferation through an interplay with stress fiber formation.