Claudin-5 expression in the vasculature of the developing chick 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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