National Xenopus Resource, Marine Biological Laboratory, Woods Hole, Massachusetts 02543, USA.
National Xenopus Resource, Marine Biological Laboratory, Woods Hole, Massachusetts 02543, USA
Cold Spring Harb Protoc. 2021 Mar 1;2021(3):pdb.prot106203. doi: 10.1101/pdb.prot106203.
Nearly a century ago, studies by Lancelot Hogben and others demonstrated that ovulation in female can be induced via injection of mammalian gonadotropins into the dorsal lymph sac, allowing for egg production throughout the year independent of the normal reproductive cycles. Hormonally induced females are capable of producing thousands of eggs in a single spawning, which can then be fertilized to generate embryos or used as a substrate for generation of egg extracts. The protocol for induction of ovulation and subsequent egg collection is straightforward and robust, yet some of its details may vary among laboratories based on prior training, availability of necessary reagents, or the experimental objectives. As the goal of this protocol is not to describe every single variation possible for acquiring eggs but to provide a simple and clear description that can be easily applied by researchers with no prior working experience with , we focus on describing the method we use at the National Resource-that is, inducing ovulation in via dorsal lymph sac injection of gonadotropic hormones and the stimulation of egg laying through application of gentle pressure to the females.
近一个世纪前,兰斯洛特·霍格本(Lancelot Hogben)等人的研究表明,通过向背部淋巴囊注射哺乳动物促性腺激素,可以诱导雌性 排卵,从而使卵子在一年中的任何时候都能不受正常生殖周期的影响而产生。通过荷尔蒙诱导的雌性能够在一次产卵中产生数千个卵子,然后这些卵子可以受精以产生胚胎,或者用作生成卵子提取物的基质。排卵诱导和随后的卵子收集的方案简单而稳健,但由于之前的培训、必要试剂的可用性或实验目标的不同,其一些细节可能在不同的实验室之间有所不同。由于本方案的目的不是描述获取卵子的每一种可能的变化,而是提供一种简单明了的描述,使没有与 合作工作经验的研究人员能够轻松应用,因此我们专注于描述我们在国家 资源中心使用的方法——即通过向背部淋巴囊注射促性腺激素诱导 排卵,并通过对雌性施加轻柔的压力来刺激产卵。