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Generation of a transgenic medaka (Oryzias latipes) strain for visualization of nuclear dynamics in early developmental stages

机译:产生转基因花aka(Oryzias latipes)菌株,用于在发育早期可视化核动力学

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摘要

In this study, we verified nuclear transport activity of an artificial nuclear localization signal (aNLS) in medaka fish (Oryzias latipes). We generated a transgenic medaka strain expresses the aNLS tagged enhanced green fluorescent protein (EGFP) driven by a medaka beta-actin promoter. The aNLS-EGFP was accumulated in the nuclei of somatic tissues and yolk nuclei of oocytes, but undetectable in the spermatozoa. The fluorescent signal was observed from immediately after fertilization by a maternal contribution. Furthermore, male and female pronuclei were visualized in fertilized eggs, and nuclear dynamics of pronuclear fusion and subsequent cleavage were captured by time-lapse imaging. In contrast, SV40NLS exhibited no activity of nuclear transport in early embryos. In conclusion, the aNLS possesses a strong nuclear localization activity and is a useful probe for fluorescent observation of the pronuclei and nuclei in early developmental stage of medaka.
机译:在这项研究中,我们验证了鱼(Oryzias latipes)中人工核定位信号(aNLS)的核转运活性。我们生成了一个转基因medaka菌株,表达了由medakaβ-actin启动子驱动的aNLS标记的增强型绿色荧光蛋白(EGFP)。 aNLS-EGFP积累在体细胞组织的核和卵母细胞的卵黄核中,但在精子中则无法检测到。受精后立即通过母亲的贡献观察到荧光信号。此外,在受精卵中可以看到雄性和雌性前核,并且通过延时成像捕获了前核融合和随后分裂的核动力学。相反,SV40NLS在早期胚胎中未显示核转运活性。总之,aNLS具有很强的核定位活性,是在med高发育初期荧光观察原核和核的有用探针。

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