Ploidy manipulation and induction of alternate cleavage patterns through inhibition of centrosome duplication in the early zebrafish embryo.
Ploidy manipulation and induction of alternate cleavage patterns through inhibition of centrosome duplication in the early zebrafish embryo.
复制标题
通过抑制早期斑马鱼胚胎中的中心体重复，对交替的切割模式进行了拼合操纵和诱导。
DOI：
10.1002/dvdy.24315
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发表时间：
2015-10
期刊：
影响因子：
--
通讯作者：
Pelegri F
中科院分区：
文献类型：
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作者：
Heier J;Takle KA;Hasley AO;Pelegri F
全基因组复制是一种有用的遗传工具，因为它允许母系后代的直接和完全的遗传纯合。热休克是斑马鱼全基因组复制的一种方法，在受精后13 - 15分钟内施加热脉冲来抑制第一次有丝分裂周期的细胞分裂。然而，热休克产生相对较低的雌雄同体产量。在第一个细胞周期（22mpf, HS2）的较晚时间点的热脉冲导致胚胎在第二个细胞周期中表现出精确的单细胞分裂停滞的高频率（bbb80 %），从而诱导全基因组复制。与单倍体生产相结合，HS2产生可存活的雌性二倍体，其产量比通过标准热冲击获得的产量高出4倍。细胞周期的延迟也会导致卵裂球分裂模式的变化，支持在下一个细胞周期中纺锤体方向的细胞质分裂。我们的研究为斑马鱼的全基因组复制、诱导雌核发生和切割模式改变提供了一种新的工具，基于细胞分裂开始前的一段时间，这段时间对温度介导的中心体复制干扰很敏感。以这一时期为目标也可能促进其他生物的遗传和发育操纵。
Whole genome duplication is a useful genetic tool because it allows immediate and complete genetic homozygosity in gynogenetic offspring. A whole genome duplication method in zebrafish, Heat Shock, involves a heat pulse in the period 13 – 15 minutes post-fertilization (mpf) to inhibit cytokinesis of the first mitotic cycle. However, Heat Shock produces a relatively low yield of gynogenotes. A heat pulse at a later time point during the first cell cycle (22 mpf, HS2) results in a high (>80%) frequency of embryos exhibiting a precise one-cell division stall during the second cell cycle, inducing whole genome duplication. Coupled with haploid production, HS2 generates viable gynogenetic diploids with yields up to 4 times higher than those achieved through standard Heat Shock. The cell cycle delay also causes blastomere cleavage pattern variations, supporting a role for cytokinesis in spindle orientation during the following cell cycle. Our studies provide a new tool for whole genome duplication, induced gynogenesis and cleavage pattern alteration in zebrafish, based on a time period prior to the initiation of cell division that is sensitive to temperature-mediated interference with centrosome duplication. Targeting of this period may also facilitate genetic and developmental manipulations in other organisms.