Generating tetraploid zebrafish by heat shock treatment and labeling microtubules of their cells in vitro

Xiudan Yuan , Yue Li , Xiaoli Hu , Wen Fu , Ruoyu Lin , Yunpeng Fan , Guangjing Zhang , Jinhui Liu , Wenbin Liu , Liangyue Peng , Yamei Xiao
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Abstract

Tetraploid is a very important experimental material, whether in scientific research or breeding practice. However, tetraploid strains are relatively rare. It was reported that chromosomes allocation disorder during cell division may lead to high embryonic mortality and low fertility in tetraploid fish. In this paper, tetraploid zebrafish is prepared by ploidy operation. And successfully establishes a tetraploid zebrafish cell line in vitro. To tracing status of microtubules, the tetraploid zebrafish cells are transfected with two plasmids of pBLK-α-tubulin-mCherry and pBLK-γ-tubulin-EGFP by electroporation in vitro. Observation shows that these transfected cells can be traced the changes of spindle and centrosomes during cell division in tetraploid cells. The results provide important material platform in vitro for studying the chromosome behavior in tetraploid cell proliferation, especially the formation mechanism of aneuploidy.

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热休克法制备四倍体斑马鱼并标记细胞微管
无论是在科学研究还是在育种实践中,四倍体都是非常重要的实验材料。然而,四倍体菌株相对罕见。据报道,四倍体鱼在细胞分裂过程中染色体分配紊乱可能导致胚胎高死亡率和低育性。本文通过倍性操作制备了四倍体斑马鱼。并在体外成功建立了四倍体斑马鱼细胞系。用pBLK-α-微管蛋白- mcherry和pBLK-γ-微管蛋白- egfp两个质粒在体外电穿孔转染四倍体斑马鱼细胞,以追踪微管状态。观察表明,这些转染后的细胞可以追踪到四倍体细胞分裂过程中纺锤体和中心体的变化。该结果为研究四倍体细胞增殖过程中染色体行为,特别是非整倍体的形成机制提供了重要的体外材料平台。
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