Csenki Zsolt, Mueller Ferenc
Faculty of Agricultural and Environmental Sciences, Department of Aquaculture, Institute of Aquaculture and Environmental Safety, Szent István University, Gödöllő, Hungary.
Institute of Cancer and Genomic Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK.
Methods Mol Biol. 2019;1920:343-352. doi: 10.1007/978-1-4939-9009-2_21.
In zebrafish and other externally developing animals, maternal effect gene products significantly contribute to embryonic and larval development. Several methods exist for both forward and reverse genetic manipulation of maternal gene products, yet it remains technically difficult to interfere with maternal gene products. Therefore, alternative methods to manipulate maternal factors in oocytes remain of interest. Here we describe a method which allows manipulation and subsequent transplantation of stage I-II follicles of zebrafish into recipient mothers where donor oocytes can develop into mature eggs and produce viable offspring. Additionally, we describe a simple microinjection protocol for injecting reporter constructs into follicles as a proxy for manipulating maternal effect genes.
在斑马鱼和其他外部发育的动物中，母体效应基因产物对胚胎和幼体发育有显著贡献。目前存在多种对母体基因产物进行正向和反向遗传操作的方法，但在技术上干扰母体基因产物仍然具有挑战性。因此，在卵母细胞中操纵母体因子的替代方法仍然备受关注。在此，我们描述了一种方法，该方法允许对斑马鱼I-II期卵泡进行操作并随后移植到受体母体中，在受体母体中供体卵母细胞能够发育成成熟卵子并产生可存活的后代。此外，我们还描述了一种简单的显微注射方案，用于将报告构建体注入卵泡，以此作为操纵母体效应基因的替代方法。
