Volume 4 Supplement 1

Proceedings of the Second International Cilia in Development and Disease Scientific Conference (2014)

Open Access

FOXA2 controls Pkd1l1 expression in the mouse node during left-right determination

  • D Leordean1,
  • D Grimes1,
  • J Keynton1,
  • J Maier2,
  • B Harfe2,
  • M Benson3,
  • A Gray3,
  • S Bhattacharya3 and
  • D Norris1
Cilia20154(Suppl 1):P37

https://doi.org/10.1186/2046-2530-4-S1-P37

Published: 13 July 2015

Objectives

The embryonic node is a ciliated pit-like structure that is important in left-right (L-R) determination. Pkd1l1, encoding a membrane-bound ciliary protein, is expressed specifically in the node and is required to establish normal L-R patterning. Given the importance of this gene, we set out to understand its regulation.

Methods

Pkd1l1 and FoxA2 expression was compared through whole-mount in situ hybridization and β-galactosidase staining in 7.5-9.5 dpc mouse embryos. Bioinformatic analysis was used to characterise FOXA2 binding regions upstream of Pkd1l1 in the human-mouse lineage. Finally, luciferase assays were used to test the functionality of these potential regulatory regions.

Results

Our expression studies indicate that a regulatory relationship might exist between FOXA2 and Pkd1l1. Furthermore, Shh -/- and FoxA2 c/c ;ShhcreER T2 embryos display abnormal Pkd1l1 expression. Indeed, previous studies identified FOXA2 binding regions upstream of Pkd1l1, in both human and mouse. Our work has revealed the two regions to be non-homologous, but conserved in the human-mouse lineage, in some cases to the binding site level. Although bioinformatic analysis predicts lower binding affinities for the conserved mouse-identified region in human and vice versa, luciferase assays indicate that the conserved regions are capable of driving gene expression.

Conclusion

Our study implicates FOXA2 in the regulation of Pkd1l1 and characterises two FOXA2 binding regions upstream of this gene. Both regions are conserved across the human-mouse lineage to varying degrees. Furthermore, in both mouse and human, the two regions are able to activate gene expression.

Authors’ Affiliations

(1)
MGU, Cilia, Development and Disease, MRC Harwell
(2)
Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida
(3)
Radcliffe Department of Medicine, University of Oxford

Copyright

© Leordean et al. 2015

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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