Moustakas-Verho Jacqueline E, McLennan Rebecca, Spengler Jennifer, Kulesa Paul M, Cebra-Thomas Judith A
Institute of Biotechnology, University of Helsinki, Helsinki, Finland.
Faculty of Biological and Environmental Sciences, University of Helsinki, Helsinki, Finland.
Methods Mol Biol. 2019;1920:247-263. doi: 10.1007/978-1-4939-9009-2_14.
Reptiles have great taxonomic diversity that is reflected in their morphology, ecology, physiology, modes of reproduction, and development. Interest in comparative and evolutionary developmental biology makes protocols for the study of reptile embryos invaluable resources. The relatively large size, seasonal breeding, and long gestation times of turtles epitomize the challenges faced by the developmental biologist. We describe protocols for the preparation of turtle embryos for ex ovo culture, electroporation, in situ hybridization, and microcomputed tomography. Because these protocols have been adapted and optimized from methods used for frog, chick, and mouse embryos, it is likely that they could be used for other reptilian species. Notes are included for alligator embryos where appropriate.
爬行动物具有极大的分类学多样性,这在它们的形态、生态、生理、繁殖方式和发育过程中都有所体现。对比较和进化发育生物学的兴趣使得研究爬行动物胚胎的实验方案成为极其宝贵的资源。海龟相对较大的体型、季节性繁殖以及较长的妊娠期,集中体现了发育生物学家所面临的挑战。我们描述了用于龟胚胎体外培养、电穿孔、原位杂交和微型计算机断层扫描的实验方案。由于这些方案是从用于青蛙、鸡和小鼠胚胎的方法改编和优化而来的,所以它们很可能也适用于其他爬行动物物种。在适当的地方还包含了针对短吻鳄胚胎的注释。