Egg Collection and In Vitro Fertilization of the Western Clawed Frog Xenopus tropicalis

Abstract

INTRODUCTIONThe eggs of Xenopus laevis have been widely used in studies investigating a variety of aspects of biology, such as control of the cell cycle, RNA processing, and the cytoskeleton. The Western clawed frog Xenopus tropicalis is likely to prove useful for such studies in the future, because of the potential to combine traditional experimental approaches with genetic analysis and the available genome sequence. The eggs of X. tropicalis are also a key starting material for transgenesis. Here, we describe a method for the routine collection of eggs from X. tropicalis, together with a method for in vitro fertilization. Very large numbers of eggs, often more than 2000, can be obtained from a single X. tropicalis female. In vitro fertilization is a valuable alternative to natural mating for generating embryos. It is particularly useful for microinjection experiments and when collecting embryos at a series of defined developmental stages (e.g., for studies of gene expression), because it produces embryos that develop synchronously during early embryonic development. The typical yield of embryos ranges from several hundred to more than 1000 per fertilization.