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Cftr controls lumen expansion and function of Kupffer's vesicle in zebrafish.

Publication ,  Journal Article
Navis, A; Marjoram, L; Bagnat, M
Published in: Development
April 2013

Regulated fluid secretion is crucial for the function of most organs. In vertebrates, the chloride channel cystic fibrosis transmembrane conductance regulator (CFTR) is a master regulator of fluid secretion. Although the biophysical properties of CFTR have been well characterized in vitro, little is known about its in vivo role during development. Here, we investigated the function of Cftr during zebrafish development by generating several cftr mutant alleles using TAL effector nucleases. We found that loss of cftr function leads to organ laterality defects. In zebrafish, left-right (LR) asymmetry requires cilia-driven fluid flow within the lumen of Kupffer's vesicle (KV). Using live imaging we found that KV morphogenesis is disrupted in cftr mutants. Loss of Cftr-mediated fluid secretion impairs KV lumen expansion leading to defects in organ laterality. Using bacterial artificial chromosome recombineering, we generated transgenic fish expressing functional Cftr fusion proteins with fluorescent tags under the control of the cftr promoter. The transgenes completely rescued the cftr mutant phenotype. Live imaging of these transgenic lines showed that Cftr is localized to the apical membrane of the epithelial cells in KV during lumen formation. Pharmacological stimulation of Cftr-dependent fluid secretion led to an expansion of the KV lumen. Conversely, inhibition of ion gradient formation impaired KV lumen inflation. Interestingly, cilia formation and motility in KV were not affected, suggesting that fluid secretion and flow are independently controlled in KV. These findings uncover a new role for cftr in KV morphogenesis and function during zebrafish development.

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Published In

Development

DOI

EISSN

1477-9129

Publication Date

April 2013

Volume

140

Issue

8

Start / End Page

1703 / 1712

Location

England

Related Subject Headings

  • Zebrafish
  • Mutagenesis
  • Morphogenesis
  • In Situ Hybridization
  • Humans
  • HEK293 Cells
  • Gene Expression Regulation, Developmental
  • Fluorescent Antibody Technique
  • Embryo, Nonmammalian
  • DNA Primers
 

Citation

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Navis, A., Marjoram, L., & Bagnat, M. (2013). Cftr controls lumen expansion and function of Kupffer's vesicle in zebrafish. Development, 140(8), 1703–1712. https://doi.org/10.1242/dev.091819
Navis, Adam, Lindsay Marjoram, and Michel Bagnat. “Cftr controls lumen expansion and function of Kupffer's vesicle in zebrafish.Development 140, no. 8 (April 2013): 1703–12. https://doi.org/10.1242/dev.091819.
Navis A, Marjoram L, Bagnat M. Cftr controls lumen expansion and function of Kupffer's vesicle in zebrafish. Development. 2013 Apr;140(8):1703–12.
Navis, Adam, et al. “Cftr controls lumen expansion and function of Kupffer's vesicle in zebrafish.Development, vol. 140, no. 8, Apr. 2013, pp. 1703–12. Pubmed, doi:10.1242/dev.091819.
Navis A, Marjoram L, Bagnat M. Cftr controls lumen expansion and function of Kupffer's vesicle in zebrafish. Development. 2013 Apr;140(8):1703–1712.

Published In

Development

DOI

EISSN

1477-9129

Publication Date

April 2013

Volume

140

Issue

8

Start / End Page

1703 / 1712

Location

England

Related Subject Headings

  • Zebrafish
  • Mutagenesis
  • Morphogenesis
  • In Situ Hybridization
  • Humans
  • HEK293 Cells
  • Gene Expression Regulation, Developmental
  • Fluorescent Antibody Technique
  • Embryo, Nonmammalian
  • DNA Primers