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Reptile Embryology and Regeneration

  • Matthew K. VickaryousEmail author
  • Emily A. B. Gilbert
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1920)

Abstract

Reptiles (lizards, snakes, turtles, and crocodilians) are becoming increasingly popular as models for developmental investigations. In this review the leopard gecko, Eublepharis macularius, is presented as a reptilian model for embryonic and tissue regeneration studies. We provide details of husbandry and breeding and discuss aspects of embryonic nutrition, egg anatomy, and sex determination. We provide comprehensive protocols for transcardial perfusion, short-term anesthesia using the injectable anesthetic Alfaxan, and full-thickness cutaneous biopsy punches, used in geckos for the study of scar-free wound healing. We also provide modifications to three popular histological techniques (whole-mount histochemistry, immunohistochemistry, and double-label immunofluorescence) and provide details on bromodeoxyuridine (BrdU) labeling and immuno-detection.

Key words

Reptilia Gecko Husbandry Embryo Sex determination Anesthesia Biopsy Perfusion whole-mount histochemistry Immunohistochemistry Immunofluorescence Proliferating cell nuclear antigen Vimentin Bromodeoxyuridine 

Notes

Acknowledgments

We thank Pat A.D. Wise for comments on the manuscript, Helen Coates and Katie McLean for assistance developing the immunohistochemical protocols, Hanna Peacock for assistance developing the biopsy protocol, and Ilinca Lungu for assistance with the photography. This work is funded by the Natural Sciences and Engineering Research Council of Canada.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Matthew K. Vickaryous
    • 1
    Email author
  • Emily A. B. Gilbert
    • 1
    • 2
  1. 1.Department of Biomedical SciencesUniversity of GuelphGuelphCanada
  2. 2.Department of Surgery, Donnelly Centre for Cellular and Biomolecular ResearchUniversity of TorontoTorontoCanada

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