Producing Xenopus Eggs and Embryos.

Xenopus laevis and Xenopus tropicalis embryos are widely used in developmental biology, genetics and disease modeling, and can be generated through in vitro fertilization (IVF), fertilizing with either fresh or cryogenically frozen sperm, or by natural matings. This chapter describes protocols developed and used successfully over two decades by the European Xenopus Resource Centre (EXRC) in Portsmouth, UK to consistently and reliably induce ovulation and generate embryos for both Xenopus laevis and Xenopus tropicalis. The following protocols describe the complete workflow for collecting Xenopus embryos, beginning from the selection of healthy Xenopus females, through to the collection of fresh sperm, hormonal induction for ovulation, egg collection via IVF and natural matings, and preparation and selection of embryos for the experimental uses described in this volume. The chapter also contains post-ovulation monitoring for both species of Xenopus females to promote a positive welfare environment for the frogs, as well as good welfare standards for the community. The following methods support consistent embryo production across a range of experimental applications, including those listed in this volume, whilst promoting best practice in husbandry and ethical use of Xenopus.