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Open Access Highly Accessed Research article

Functional conservation of a forebrain enhancer from the elephant shark (Callorhinchus milii ) in zebrafish and mice

Ryan B MacDonald1, Mélanie Debiais-Thibaud1, Kyle Martin13, Luc Poitras1, Boon-Hui Tay2, Byrappa Venkatesh2 and Marc Ekker1*

Author Affiliations

1 Center for Advanced Research in Environmental Genomics, Department of Biology, University of Ottawa, Ottawa, ON, K1N 6N5, Canada

2 Institute of Molecular and Cell Biology, A*STAR, Biopolis, 138673, Singapore

3 Department of Zoology, Oxford University, Oxford, UK

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BMC Evolutionary Biology 2010, 10:157  doi:10.1186/1471-2148-10-157

Published: 26 May 2010

Abstract

Background

The phylogenetic position of the elephant shark (Callorhinchus milii ) is particularly relevant to study the evolution of genes and gene regulation in vertebrates. Here we examine the evolution of Dlx homeobox gene regulation during vertebrate embryonic development with a particular focus on the forebrain. We first identified the elephant shark sequence orthologous to the URE2 cis -regulatory element of the mouse Dlx1/Dlx2 locus (herein named CmURE2). We then conducted a comparative study of the sequence and enhancer activity of CmURE2 with that of orthologous regulatory sequences from zebrafish and mouse.

Results

The CmURE2 sequence shows a high percentage of identity with its mouse and zebrafish counterparts but is overall more similar to mouse URE2 (MmURE2) than to zebrafish URE2 (DrURE2). In transgenic zebrafish and mouse embryos, CmURE2 displayed enhancer activity in the forebrain that overlapped with that of DrURE2 and MmURE2. However, we detected notable differences in the activity of the three sequences in the diencephalon. Outside of the forebrain, CmURE2 shows enhancer activity in areas such as the pharyngeal arches and dorsal root ganglia where its' counterparts are also active.

Conclusions

Our transgenic assays show that part of the URE2 enhancer activity is conserved throughout jawed vertebrates but also that new characteristics have evolved in the different groups. Our study demonstrates that the elephant shark is a useful outgroup to study the evolution of regulatory mechanisms in vertebrates and to address how changes in the sequence of cis -regulatory elements translate into changes in their regulatory activity.