Elsevier

Gene Expression Patterns

Volume 12, Issues 3–4, March–April 2012, Pages 123-129
Gene Expression Patterns

Claudin-5 expression in the vasculature of the developing chick embryo

https://doi.org/10.1016/j.gep.2012.01.005Get rights and content

Abstract

The claudin family of proteins are integral components of tight junctions and are responsible for determining the ion specificity and permeability of paracellular transport within epithelial and endothelial cell layers. Studies in human, mouse, Xenopus, and zebrafish have shown that only a limited number of claudins are expressed in endothelial cells. Here, we report the expression pattern of Claudin-5 during chick development. Between HH stage 4 and 6 Claudin-5 expression was observed exclusively in extraembryonic tissue. Claudin-5 expression was not observed in the embryo until HH stage 8, coincident with the onset of embryonic vascularization. Claudin-5 expression was maintained in the developing vasculature in the embryonic and extraembryonic tissue throughout organogenesis (HH stage 19–35), including the vasculature of the ectoderm and of organs derived from the mesoderm and endoderm lineages. These data describe a conserved expression pattern for Claudin-5 in the endothelial tight junction barrier and is the first report of the onset of Claudin-5 expression in a vertebrate embryo.

Highlights

Claudin-5 is the only endothelial claudin expressed during chick development. ► Functional amino acids are conserved in chick Claudin-5. ► Onset of embryonic expression in the vasculature is HH stage 8. ► Claudin-5 is highly expressed in vasculature throughout the body.

Section snippets

Claudin-5 is evolutionarily conserved in the chick

To determine the similarity between Claudin-5 in the chick and other vertebrates, we performed phylogenetic analyses using the human, mouse, Xenopus, and chick Claudin-5 protein sequence (Fig. 1A and B). Examination of the entire protein coding sequence reveals that chick Claudin-5 is most closely related to Xenopus (73%) and shows 70% sequence identity to both the human and mouse orthologs. These levels of identity are higher in known functional domains of the protein. The chick and human are

RT-PCR and cDNA cloning of chick Claudin-5

Fertilized eggs were obtained from Couvoir Simetin Hatchery (Mirabel, Quebec, Canada) and incubated at 38.5 °C until the appropriate age was reached. Embryos were handled according to the Canadian Council on Animal Care guidelines and were staged using Hamburger and Hamilton (HH) staging criteria (Hamburger and Hamilton, 1951). Whole HH stage 4 embryos were dissected and flash frozen in liquid nitrogen. Poly A+ mRNA was isolated using the MicroFast Track 2.0 kit (Invitrogen, Carlsbad, CA) and

Acknowledgements

We would like to thank members of the Ryan lab, I. Gupta, and L. Jerome-Majewska for helpful discussions and comments. M.M.C. is the recipient of a doctoral studentship from the Fonds de la Recherche en Santé du Quebec (FRSQ) and A.I.B. is the recipient of a studentship from the Foundation of Stars. This work was supported by a Natural Sciences and Engineering Research of Canada Discovery Grant and Canadian Institutes of Health Research Operating Grant to A.K.R. A.K.R. is a member of the

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