Sodium/hydrogen‐exchanger‐2 modulates colonocyte lineage differentiation

AbstractAimThe sodium/hydrogen exchanger 2 (NHE2) is an intestinal acid extruder with crypt‐predominant localization and unresolved physiological significance. Our aim was to decipher its role in colonic epithelial cell proliferation, differentiation and electrolyte transport.MethodsAlterations induced by NHE2‐deficiency were addressed in murine nhe2−/− and nhe2+/+ colonic crypts and colonoids, and NHE2‐knockdown and control Caco2Bbe cells using pH‐fluorometry, gene expression analysis and immunofluorescence.ResultspHi‐measurements along the colonic cryptal axis revealed significantly decreased intracellular pH (pHi) in the middle segment of nhe2−/− compared to nhe2+/+ crypts. Increased Nhe2 mRNA expression was detected in murine colonoids in the transiently amplifying/progenitor cell stage (TA/PE). Lack of Nhe2 altered the differentiation programme of colonic epithelial cells with reduced expression of absorptive lineage markers alkaline phosphatase (iAlp), Slc26a3 and transcription factor hairy and enhancer‐of‐split 1 (Hes1), but increased expression of secretory lineage markers Mucin 2, trefoil factor 3 (Tff3), enteroendocrine marker chromogranin A and murine atonal homolog 1 (Math1). Enterocyte differentiation was found to be pHi dependent with acidic pHi reducing, and alkaline pHi stimulating the expression of enterocyte differentiation markers in Caco2Bbe cells. A thicker mucus layer, longer crypts and an expanded brush border membrane zone of sodium/hydrogen exchanger 3 (NHE3) abundance may explain the lack of inflammation and the normal fluid absorptive rate in nhe2−/− colon.ConclusionsThe results suggest that NHE2 expression is activated when colonocytes emerge from the stem cell niche. Its activity increases progenitor cell pHi and thereby supports absorptive enterocyte differentiation.