Unilateral Gonadectomy in Adult Male Xenopus laevis

In vitro fertilization is a reliable laboratory technique for generating large numbers of synchronously developing embryos. This synchronicity enhances Xenopus laevis as a powerful model organism for studying vertebrate embryonic development, molecular, cellular, and disease mechanisms. In conventional in vitro fertilization protocols for X. laevis, male frogs are euthanized, and both testes are harvested. In order to reduce the number of required males, a procedure was developed for unilateral gonadectomy in male X. laevis, which allows sequential use of male frogs' gonads for in vitro fertilizations. This approach has been successfully used to generate embryos, yielding equally good fertilization outcomes with sperm from either the surgically removed or the contralateral testis. The surgical process involves the administration of MS-222 anesthesia, use of Meloxicam and Bupivacaine analgesia, and post-operative care and monitoring, thus allowing reduction in animal use and supporting best practices in animal care across multiple experimental conditions and laboratory setting.