Details
| Original language | English |
|---|---|
| Article number | e13774 |
| Journal | Acta Physiologica |
| Volume | 234 |
| Issue number | 3 |
| Publication status | Published - 5 Jan 2022 |
| Externally published | Yes |
Abstract
Aim: The 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. Methods: Alterations 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. Results: pHi-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. Conclusions: The 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.
Keywords
- colonoids, intestinal differentiation, intestinal electrolyte transport, NHE2, pH regulation, sodium/hydrogen exchange
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Physiology
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In: Acta Physiologica, Vol. 234, No. 3, e13774, 05.01.2022.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Sodium/hydrogen-exchanger-2 modulates colonocyte lineage differentiation
AU - Nikolovska, Katerina
AU - Cao, Li
AU - Hensel, Inga
AU - Di Stefano, Gabriella
AU - Seidler, Anna Elisabeth
AU - Zhou, Kunyan
AU - Qian, Jiajie
AU - Singh, Anurag Kumar
AU - Riederer, Brigitte
AU - Seidler, Ursula
N1 - Publisher Copyright: © 2022 The Authors. Acta Physiologica published by John Wiley & Sons Ltd on behalf of Scandinavian Physiological Society
PY - 2022/1/5
Y1 - 2022/1/5
N2 - Aim: The 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. Methods: Alterations 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. Results: pHi-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. Conclusions: The 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.
AB - Aim: The 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. Methods: Alterations 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. Results: pHi-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. Conclusions: The 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.
KW - colonoids
KW - intestinal differentiation
KW - intestinal electrolyte transport
KW - NHE2
KW - pH regulation
KW - sodium/hydrogen exchange
UR - http://www.scopus.com/inward/record.url?scp=85123503752&partnerID=8YFLogxK
U2 - 10.1111/apha.13774
DO - 10.1111/apha.13774
M3 - Article
C2 - 34985202
AN - SCOPUS:85123503752
VL - 234
JO - Acta Physiologica
JF - Acta Physiologica
SN - 1748-1708
IS - 3
M1 - e13774
ER -