Olson David J, Hulstrand Alissa M, Houston Douglas W
Department of Biology, The University of Iowa, Iowa City, IA, USA.
Methods Mol Biol. 2012;917:167-82. doi: 10.1007/978-1-61779-992-1_10.
The ability to inhibit the activity of maternally stored gene products in Xenopus has led to numerous insights into early developmental mechanisms. Oocytes can be cultured and manipulated in vitro and then implanted into the body cavity of a host female to make them competent for fertilization. Here, we summarize the methods for obtaining, culturing, and fertilizing Xenopus oocytes, with the goal of inhibiting maternal gene function through antisense oligonucleotide-mediated mRNA knock-down. We describe a simplified technique for implanting donor oocytes into host females using intraperitoneal injection. Also, we present optimized methods for performing the host-transfer procedure with Xenopus tropicalis oocytes.
在非洲爪蟾中抑制母源储存基因产物活性的能力,为深入了解早期发育机制带来了诸多见解。卵母细胞可以在体外进行培养和操作,然后植入宿主雌性的体腔中,使其具备受精能力。在这里,我们总结了获取、培养和使非洲爪蟾卵母细胞受精的方法,目的是通过反义寡核苷酸介导的mRNA敲低来抑制母源基因功能。我们描述了一种使用腹腔注射将供体卵母细胞植入宿主雌性的简化技术。此外,我们还介绍了用热带爪蟾卵母细胞进行宿主转移程序的优化方法。