Noramly Selina, Zimmerman Lyle, Cox Amanda, Aloise Renee, Fisher Marilyn, Grainger Robert M
Department of Biology, University of Virginia, Gilmer Hall, Charlottesville, VA 22904, USA.
Mech Dev. 2005 Mar;122(3):273-87. doi: 10.1016/j.mod.2004.11.001.
In the rapidly developing, diploid amphibian Xenopus tropicalis, genetics can be married to the already powerful tools of the amphibian system to overcome a disability that has hampered Xenopus laevis as a model organism: the difficulties inherent in conducting genetic analyses in a tetraploid organism with a longer generation time. We describe here a gynogenetic screen to uncover naturally occurring recessive mutations in wild X. tropicalis populations, a procedure that is both faster and easier than conventional genetic screens traditionally employed in model organisms to dissect early developmental pathways. During the first round of our screen, gynogenetic diploids from over 160 females comprising four different wild-caught populations were examined. Forty-two potential mutant phenotypes were isolated during this round of gynogenesis. From this group, we describe 10 lines that have genetically heritable recessive mutations. A wide range of developmental defects were obtained in this screen, encompassing effects limited to individual organs as well phenotypes characterized by more global changes in tadpole body morphology. The frequency of recessive mutations detected in our screen appears lower than that seen in other vertebrate genetic screens, but given constraints on the screening procedure used here, is likely to be consistent with rates seen in other animals, and clearly illustrates how wild-caught animals can be a productive source of developmental mutations for experimental study. The development of genetic strategies for the Xenopus system, together with new genomic resources, existing technologies for transgenesis, and other means for manipulating gene expression, as well as the power of performing embryonic manipulations, will provide an impressive set of tools for resolving complex cell and developmental phenomena in the future.
在快速发展的二倍体两栖动物热带爪蟾中,遗传学可以与两栖动物系统中现有的强大工具相结合,以克服阻碍非洲爪蟾成为模式生物的一个障碍:在四倍体生物中进行遗传分析存在困难,且其世代时间较长。我们在此描述一种雌核发育筛选方法,以揭示野生热带爪蟾种群中自然发生的隐性突变,该方法比传统用于剖析早期发育途径的模式生物遗传筛选方法更快、更简便。在我们第一轮筛选中,检查了来自四个不同野生捕获种群的160多名雌性的雌核发育二倍体。在这一轮雌核发育过程中分离出42种潜在的突变表型。从这一组中,我们描述了10个具有可遗传隐性突变的品系。在这个筛选中获得了广泛的发育缺陷,包括仅限于单个器官的影响以及以蝌蚪身体形态更全面变化为特征的表型。我们筛选中检测到的隐性突变频率似乎低于其他脊椎动物遗传筛选中观察到的频率,但考虑到这里使用的筛选程序的限制,很可能与其他动物中观察到的频率一致,并且清楚地说明了野生捕获的动物如何能够成为用于实验研究的发育突变的丰富来源。爪蟾系统遗传策略的发展,连同新的基因组资源、现有的转基因技术以及其他操纵基因表达的手段,以及进行胚胎操作的能力,将为未来解析复杂的细胞和发育现象提供一套令人印象深刻的工具。