The Zebrafish curly fry Is Required for Proper Centrosome and Mitotic Spindle Assembly.
The Zebrafish curly fry Is Required for Proper Centrosome and Mitotic Spindle Assembly.
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斑马鱼卷曲鱼苗是正确的中心体和有丝分裂纺锤体组装所必需的。
DOI：
10.1089/zeb.2017.1427
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发表时间：
2017
期刊：
影响因子：
2
通讯作者：
Kuwada,JohnY
中科院分区：
文献类型：
--
作者：
Song,MiHye;Medley,JeffreyC;Kuwada,JohnY
斑马鱼卷曲鱼苗（cfy）突变导致有丝分裂指数的急剧增加和神经管形成期间开始的细胞死亡。突变表型是细胞自主的，并不导致缺陷的顶端/基底极性内的神经上皮。有丝分裂指数的增加可能是由于增殖增加或有丝分裂中的细胞周期停滞。分析cfy胚胎以检查这两种可能性。通过用BrdU和抗磷酸化组蛋白3脉冲标记胚胎，并通过荧光激活细胞分选术检查DNA含量，我们发现cfy突变体没有表现出增殖的增加，但有丝分裂停滞的细胞数量显著增加。此外，体内延时显微镜证实，绝大多数分裂细胞在有丝分裂过程中停滞，这些有丝分裂停滞的细胞在胚胎中死亡。最后，免疫染色和共聚焦显微镜infymutant胚胎显示，有丝分裂细胞的突变体含有异常的中心体，往往表现出单极纺锤体，从而导致有丝分裂细胞周期停滞。我们的研究结果表明，fy基因是中心体组装和有丝分裂纺锤体形成所必需的，因此对正常细胞分裂至关重要。
The zebrafishcurly fry(cfy) mutation leads to a dramatic increase in mitotic index and cell death starting during neural tube formation. The mutant phenotype is cell autonomous and does not result from defects in apical/basal polarity within the neuroepithelium. The increase in mitotic index could be due to increased proliferation or cell cycle arrest in mitosis.cfyembryos were analyzed to examine these two possibilities. By labeling embryos with a pulse of BrdU and anti-phospho-histone 3 and examining the DNA content by fluorescence activated cell sorting, we show thatcfymutants exhibit no increase in proliferation, but a significant increase in the number of cells arrested in mitosis. Furthermore, time-lapse microscopyin vivoconfirmed that a great majority of dividing cells arrest during mitosis and that these mitotically arrested cells die incfyembryos. Finally, immunostaining and confocal microscopy incfymutant embryos revealed that mitotic cells in mutants contain aberrant centrosomes and often exhibit monopolar spindles, thereby leading to mitotic cell cycle arrest. Our results suggest that thecfygene is required for proper centrosome assembly and mitotic spindle formation, therefore critical for normal cell division.