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Carbachol-induced colonic mucus formation requires transport via NKCC1, K+ channels and CFTR

  • Integrative physiology
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Abstract

The colonic mucosa protects itself from the luminal content by secreting mucus that keeps the bacteria at a distance from the epithelium. For this barrier to be effective, the mucus has to be constantly replenished which involves exocytosis and expansion of the secreted mucins. Mechanisms involved in regulation of mucus exocytosis and expansion are poorly understood, and the aim of this study was to investigate whether epithelial anion secretion regulates mucus formation in the colon. The muscarinic agonist carbachol was used to induce parallel secretion of anions and mucus, and by using established inhibitors of ion transport, we studied how inhibition of epithelial transport affected mucus formation in mouse colon. Anion secretion and mucin exocytosis were measured by changes in membrane current and epithelial capacitance, respectively. Mucus thickness measurements were used to determine the carbachol effect on mucus growth. The results showed that the carbachol-induced increase in membrane current was dependent on NKCC1 co-transport, basolateral K+ channels and Cftr activity. In contrast, the carbachol-induced increase in capacitance was partially dependent on NKCC1 and K+ channel activity, but did not require Cftr activity. Carbachol also induced an increase in mucus thickness that was inhibited by the NKCC1 blocker bumetanide. However, mice that lacked a functional Cftr channel did not respond to carbachol with an increase in mucus thickness, suggesting that carbachol-induced mucin expansion requires Cftr channel activity. In conclusion, these findings suggest that colonic epithelial transport regulates mucus formation by affecting both exocytosis and expansion of the mucin molecules.

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Acknowledgments

This work was supported by the Swedish Research Council (nos. 7461, 8288, 21027, 21372, 20693, 2195501, 2009-2420, 521-2011-2370 and 342-2004-4434), The Knut and Alice Wallenberg Foundation, IngaBritt and Arne Lundberg Foundation, Sahlgren’s University Hospital (LUA-ALF), Wilhelm and Martina Lundgren’s Foundation, Åke Wibergs Foundation, Torsten and Ragnar Söderbergs Foundations, The Swedish Foundation for Strategic Research-The Mucosal Immunobiology and Vaccine Center (MIVAC) and the Mucus-Bacteria-Colitis Center (MBC) of the Innate Immunity Program. We acknowledge Dr. Anna Velcich for the Muc2−/− animals and Maria Sapnara for the technical assistance.

Ethical standards

All experiments were performed according to the ethical guidelines of the University of Gothenburg.

Conflict of interest

The authors declare that they have no conflicts of interest.

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Authors and Affiliations

  1. Department of Medical Biochemistry, University of Gothenburg, Medicinaregatan 9A, Box 440, Gothenburg, 405 30, Sweden

    Jenny K. Gustafsson, Sara K. Lindén, Ala H. Alwan & Gunnar C. Hansson

  2. Department of Cell Biology, Erasmus MC, Rotterdam, The Netherlands

    Bob J. Scholte

  3. Department of Internal Medicine, University of Gothenburg, Gothenburg, Sweden

    Henrik Sjövall

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  1. Jenny K. Gustafsson
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Correspondence to Jenny K. Gustafsson.

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Gustafsson, J.K., Lindén, S.K., Alwan, A.H. et al. Carbachol-induced colonic mucus formation requires transport via NKCC1, K+ channels and CFTR. Pflugers Arch - Eur J Physiol 467, 1403–1415 (2015). https://doi.org/10.1007/s00424-014-1595-y

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