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Pflügers Archiv - European Journal of Physiology

, Volume 470, Issue 9, pp 1335–1348 | Cite as

Expression and function of Anoctamin 1/TMEM16A calcium-activated chloride channels in airways of in vivo mouse models for cystic fibrosis research

  • Anne Hahn
  • Johanna J. Salomon
  • Dominik Leitz
  • Dennis Feigenbutz
  • Lisa Korsch
  • Ina Lisewski
  • Katrin Schrimpf
  • Pamela Millar-Büchner
  • Marcus A. Mall
  • Stephan FringsEmail author
  • Frank Möhrlen
Molecular and cellular mechanisms of disease
Part of the following topical collections:
  1. Topical Collection: Molecular and cellular mechanisms of disease

Abstract

Physiological processes of vital importance are often safeguarded by compensatory systems that substitute for primary processes in case these are damaged by gene mutation. Ca2+-dependent Cl secretion in airway epithelial cells may provide such a compensatory mechanism for impaired Cl secretion via cystic fibrosis transmembrane conductance regulator (CFTR) channels in cystic fibrosis (CF). Anoctamin 1 (ANO1) Ca2+-gated Cl channels are known to contribute to calcium-dependent Cl secretion in tracheal and bronchial epithelia. In the present study, two mouse models of CF were examined to assess a potential protective function of Ca2+-dependent Cl secretion, a CFTR deletion model (cftr−/−), and a CF pathology model that overexpresses the epithelial Na+ channel β-subunit (βENaC), which is encoded by the Scnn1b gene, specifically in airway epithelia (Scnn1b-Tg). The expression levels of ANO1 were examined by mRNA and protein content, and the channel protein distribution between ciliated and non-ciliated epithelial cells was analyzed. Moreover, Ussing chamber experiments were conducted to compare Ca2+-dependent Cl secretion between wild-type animals and the two mouse models. Our results demonstrate that CFTR and ANO1 channels were co-expressed with ENaC in non-ciliated cells of mouse tracheal and bronchial epithelia. Ciliated cells did not express these proteins. Despite co-localization of CFTR and ANO1 in the same cell type, cells in cftr−/− mice displayed no altered expression of ANO1. Similarly, ANO1 expression was unaffected by βENaC overexpression in the Scnn1b-Tg line. These results suggest that the CF-related environment in the two mouse models did not induce ANO1 overexpression as a compensatory system.

Keywords

Airway epithelium Chloride secretion Cystic fibrosis Anoctamin TMEM16A Mouse models 

Notes

Acknowledgements

We thank Dr. Alexei Diakov for kindly providing the βENaC antibody.

Funding information

This project was supported through a grant to AH by the Studienstiftung des deutschen Volkes and in part by the German Ministry for Education and Research (82DZL00401, 82DZL0040A1) to MAM.

Compliance with ethical standards

All experiments were approved by the Regierungspräsidium Karlsruhe and were conducted in agreement with national and international guidelines.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Anne Hahn
    • 1
  • Johanna J. Salomon
    • 2
  • Dominik Leitz
    • 2
  • Dennis Feigenbutz
    • 1
  • Lisa Korsch
    • 1
  • Ina Lisewski
    • 1
  • Katrin Schrimpf
    • 1
  • Pamela Millar-Büchner
    • 2
  • Marcus A. Mall
    • 2
    • 3
    • 4
  • Stephan Frings
    • 1
    Email author
  • Frank Möhrlen
    • 1
  1. 1.Department of Animal Molecular Physiology, Centre of Organismal StudiesUniversity of HeidelbergHeidelbergGermany
  2. 2.Department of Translational Pulmonology, Translational Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL)University of HeidelbergHeidelbergGermany
  3. 3.Department of Pediatric Pulmonology and ImmunologyCharité-Universitätsmedizin BerlinBerlinGermany
  4. 4.Berlin Institute of Health (BIH)BerlinGermany

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