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Cited by 58Ion Transport and Channel RegulationDigestive system and related healthGastrointestinal motility and disorders

Molecular Basis and Differentiation-Associated Alterations of Anion Secretion in Human Duodenal Enteroid Monolayers

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Yin, Jianyi, Tse, Chung-Ming, Avula, Leela Rani, Singh, Varsha, Foulke-Abel, Jennifer, de Jonge, Hugo R., Donowitz, Mark·2018·Cellular and Molecular Gastroenterology and Hepatology
DOI 10.1016/j.jcmgh.2018.02.002PMC PMC6009799
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Abstract

Background & Aims Human enteroids present a novel tool to study human intestinal ion transport physiology and pathophysiology. The present study describes the contributions of Cl - and HCO 3 - secretion to total cyclic adenosine monophosphate (cAMP)-stimulated electrogenic anion secretion in human duodenal enteroid monolayers and the relevant changes after differentiation. Methods Human duodenal enteroids derived from 4 donors were grown as monolayers and differentiated by a protocol that includes the removal of Wnt3A, R-spondin1, and SB202190 for 5 days. The messenger RNA level and protein expression of selected ion transporters and carbonic anhydrase isoforms were determined by quantitative real-time polymerase chain reaction and immunoblotting, respectively. Undifferentiated and differentiated enteroid monolayers were mounted in the Ussing chamber/voltage-current clamp apparatus, using solutions that contained as well as lacked Cl - and HCO 3 - /CO 2 , to determine the magnitude of forskolin-induced short-circuit current change and its sensitivity to specific inhibitors that target selected ion transporters and carbonic anhydrase(s). Results Differentiation resulted in a significant reduction in the messenger RNA level and protein expression of cystic fibrosis transmembrane conductance regulator, (CFTR) Na + /K + /2Cl - co-transporter 1 (NKCC1), and potassium channel, voltage gated, subfamily E, regulatory subunit 3 (KCNE3); and, conversely, increase of down-regulated-in-adenoma (DRA), electrogenic Na + /HCO 3 - co-transporter 1 (NBCe1), carbonic anhydrase 2 (CA2), and carbonic anhydrase 4 (CA4). Both undifferentiated and differentiated enteroids showed active cAMP-stimulated anion secretion that included both Cl - and HCO 3 - secretion as the magnitude of total active anion secretion was reduced after the removal of extracellular Cl - or HCO 3 - /CO 2 . The magnitude of total anion secretion in differentiated enteroids was approximately 33% of that in undifferentiated enteroids, primarily owing to the reduction in Cl - secretion with no significant change in HCO 3 - secretion. Anion secretion was consistently lower but detectable in differentiated enteroids compared with undifferentiated enteroids in the absence of extracellular Cl - or HCO 3 - /CO 2 . Inhibiting CFTR, NKCC1, carbonic anhydrase(s), cAMP-activated K + channel(s), and Na + /K + -adenosine triphosphatase reduced cAMP-stimulated anion secretion in both undifferentiated and differentiated enteroids. Conclusions Human enteroids recapitulate anion secretion physiology of small intestinal epithelium. Enteroid differentiation is associated with significant alterations in the expression of several ion transporters and carbonic anhydrase isoforms, leading to a reduced but preserved anion secretory phenotype owing to markedly reduced Cl - secretion but no significant change in HCO 3 - secretion.

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  • Compared to undifferentiated organoids, expression of the intestinal stem cell marker LGR5 and the proliferation marker MKI67 was substantially decreased in differentiated monolayers.

    “Compared to undifferentiated organoids, expression of the intestinal stem cell marker LGR5 and the proliferation marker MKI67 was substantially decreased in differentiated monolayers (Figure 1C), in accordance with an earlier study performed in human duodenal organoid-derived monolayers [16].”

    From Regulation of Enterocyte Brush Border Membrane Primary Na-Absorptive Transporters in Human Intestinal Organoid-Derived Monolayers (2024) · cites this paper as [16]

  • Human intestinal organoid-derived monolayers have been successfully used to study host-pathogen interactions, epithelial barrier function, and various transport processes including secretion of hormones, growth factors, and ions, and absorption of glucose.

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    From Regulation of Enterocyte Brush Border Membrane Primary Na-Absorptive Transporters in Human Intestinal Organoid-Derived Monolayers (2024) · cites this paper as [16]

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Cited by

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Authors
Yin, J., Tse, C.-M., Avula, L. R., Singh, V., Foulke-Abel, J., de Jonge, H. R., & Donowitz, M.
Affiliations
Johns Hopkins University (US)
Corresponding author
Donowitz, Mark
ORCID
Venue
Cellular and Molecular Gastroenterology and Hepatology
Publisher
Elsevier BV
Published
2018
Volume
5
Issue
4
Pages
591-609
ISSN
2352-345X (ISSN-L)
DOI
10.1016/j.jcmgh.2018.02.002
PMC
PMC6009799
OpenAlex
W2801507037
Cited by
58
References
3
Citations by year
2026: 5 · 2025: 7 · 2024: 7 · 2023: 5 · 2022: 6 · 2021: 6 · 2020: 9 · 2019: 9 · 2018: 4
FWCI
2.49
Type
journal article
Language
en
Open access
Licence
Locations
  • Cellular and Molecular Gastroenterology and Hepatology (publishedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • EUR Research Repository (Erasmus University Rotterdam) (publishedVersion, other-oa)
  • EUR Research Repository (Erasmus University Rotterdam) (submittedVersion, other-oa)
  • RePub (Erasmus University Rotterdam) (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
National Institutes of Health (RO1 DK26523, RO1 DK61765, P01 DK072084, P30 DK089502, UO1AI125181, UH3TR00003, U18TR000552, R24 DK99803)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Digestive system and related health
  • Health Sciences › Medicine › Gastroenterology › Gastrointestinal motility and disorders
Keywords
anion secretion, cyclic AMP, short-circuit current, carbonic anhydrase, Na+/K+-ATPase, ion transporters, duodenal epithelium

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