Pediatric Genomics Discovery Program, Department of Pediatrics and Genetics, Yale University School of Medicine, New Haven, Connecticut 06510, USA
Cold Spring Harb Protoc. 2022 Apr 1;2022(4):Pdb.prot106609. doi: 10.1101/pdb.prot106609.
is a powerful model system for cell and developmental biology in part because frogs produce thousands of eggs and embryos year-round. Natural matings are a simple and common method to obtain embryos for injection or other experimental use or to raise to adulthood. This method does not require sacrificing a male as in vitro fertilization (IVF) does. Male and female frogs are injected with an ovulation hormone, placed together in a mating bucket, and left for 4-6 h or overnight to mate. Embryos are then collected, treated with cysteine to remove the sticky jelly coat, and used for injections and/or raised to later stages or adulthood. For embryos raised past free-swimming stages, the cysteine step can optionally be skipped, and tadpoles can be allowed to hatch naturally from the jelly coat. Although there are many similarities between natural mating protocols for and , there are key differences such as hormone dosage, timing of ovulation, and embryo incubation temperature. Here we provide a specific protocol for inducing natural matings in .
是细胞和发育生物学的强大模型系统,部分原因是青蛙可以全年产生数千个卵子和胚胎。自然交配是一种简单而常见的方法,可以获得用于注射或其他实验用途的胚胎,或饲养至成年。与体外受精 (IVF) 不同,这种方法不需要牺牲雄性。向雄性和雌性青蛙注射排卵激素,将它们放在交配桶中,放置 4-6 小时或过夜以交配。然后收集胚胎,用半胱氨酸处理以去除粘性的果冻层,用于注射和/或培养到后期阶段或成年期。对于培养到自由游泳阶段之后的胚胎,可以选择跳过半胱氨酸步骤,让蝌蚪自然从果冻层孵化。尽管 和 之间的自然交配协议有许多相似之处,但也存在关键差异,例如激素剂量、排卵时间和胚胎孵化温度。在这里,我们提供了一个在 中诱导自然交配的具体方案。