Urushibata Hirotaro, Takahashi Eisuke, Shimizu Yu, Miyazaki Toshiya, Fujimoto Takafumi, Arai Katsutoshi, Yamaha Etsuro
Nanae Freshwater Station, Field Science Center for Northern Biosphere, Hokkaido University,Hakodate, Hokkaido, Japan.
Int J Dev Biol. 2019;63(11-12):597-604. doi: 10.1387/ijdb.190144hu.
The goldfish (Carassius auratus auratus) is a useful species for embryonic micromanipulations because of its large egg size and wide temperature tolerance. Here, we describe in detail the rate of development and morphological characteristics of goldfish embryos incubated at temperatures between 10 °C and 30 °C. The cleavage speed increased rapidly as temperature increased. Synchronized cell divisions occurred at 131 min intervals at 10 °C, at 33 min intervals at 20 °C, and at 19 min intervals at 30 °C during the cleavage period. The rate of hatched abnormal embryos significantly increased at temperatures of 26 °C and above, while there was no change in the number of abnormal embryos at temperatures less than 24 °C. Moreover, the blastomeres around the center of the blastodisc rose in the direction of the animal pole at temperatures less than 14 °C. At the lower temperatures, clusters of maternally-supplied germplasm were visualized both at the ends of the first three cleavage furrows and at the border between the lower and upper tiers at the 16- to 32-cell stage, with injection of artificial mRNA and vasa in situ hybridization. This study showed that temperature affects not only developmental speed but also the shape of the blastodisc and the distribution of maternally-supplied materials in the blastodisc. By controlling the temperature, it is possible for researchers to prepare many stages of embryos and shapes of the blastodisc from a single batch of eggs.
金鱼(Carassius auratus auratus)因其卵体积大且温度耐受性广,是胚胎显微操作的有用物种。在此,我们详细描述了在10℃至30℃之间孵化的金鱼胚胎的发育速率和形态特征。随着温度升高,卵裂速度迅速增加。在卵裂期,10℃时细胞同步分裂间隔为131分钟,20℃时为33分钟,30℃时为19分钟。在26℃及以上温度时,孵化出的异常胚胎比例显著增加,而在低于24℃的温度下,异常胚胎数量没有变化。此外,在低于14℃的温度下,胚盘中心周围的卵裂球向动物极方向上升。在较低温度下,通过注射人工mRNA和原位杂交检测vas a,在前三次卵裂沟的末端以及16至32细胞期上下层之间的边界处都能看到母源种质簇。本研究表明,温度不仅影响发育速度,还影响胚盘的形状以及胚盘中母源物质的分布。通过控制温度,研究人员可以从一批卵中制备出许多胚胎阶段和胚盘形状。