Csenki Zsolt, Zaucker Andreas, Kovács Balázs, Hadzhiev Yavor, Hegyi Arpád, Lefler Katalin-Kinga, Müller Tamás, Kovács Robert, Urbányi Béla, Váradi László, Müller Ferenc
Department of Fish Culture, Faculty of Agricultural and Environmental Sciences, Szent István University, Gödöllo, Hungary.
Int J Dev Biol. 2010;54(4):585-9. doi: 10.1387/ijdb.082786zc.
Maternal gene products drive early embryogenesis almost exclusively until the mid blastula transition (MBT) in many animal models including fish. However, the maternal contribution to embryogenesis does not stop at MBT, but continues to be an essential regulator of key developmental processes. The extent to which maternal effects contribute to embryonic and larval development is hard to estimate due to the technical difficulty of interfering with maternal gene products by conventional forward and reverse genetic tools. Therefore, novel methods to manipulate maternal factors in oocytes need to be developed. Here, we provide a proof of principle protocol for transplanting stage I-II zebrafish follicles into recipient mothers where donor stage I oocytes can develop to stage IV in 2 weeks and in 3 weeks they develop into mature eggs and produce viable offspring. Moreover, we show that simple microinjection of stage I-II follicles with RNA results in reporter gene expression in oocytes and paves the way for developing tools for interfering with maternal gene activity. This early stage oocyte transplantation protocol provides a means to study cellular and molecular aspects of oocyte development in the zebrafish.
在包括鱼类在内的许多动物模型中,母源基因产物几乎完全驱动早期胚胎发育,直至中囊胚转换(MBT)。然而,母源对胚胎发育的贡献并非在MBT时停止,而是继续作为关键发育过程的重要调节因子。由于使用传统正向和反向遗传工具干扰母源基因产物存在技术困难,母源效应在胚胎和幼体发育中的作用程度难以估计。因此,需要开发新的方法来操纵卵母细胞中的母源因子。在此,我们提供了一个原理验证方案,即将I-II期斑马鱼卵泡移植到受体母鱼中,供体I期卵母细胞在2周内可发育到IV期,3周内发育成成熟卵子并产生可存活的后代。此外,我们表明,对I-II期卵泡简单地进行RNA显微注射会导致卵母细胞中报告基因表达,为开发干扰母源基因活性的工具铺平了道路。这种早期卵母细胞移植方案为研究斑马鱼卵母细胞发育的细胞和分子方面提供了一种手段。