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Methods Cell Biol. 2018;145:237-248. doi: 10.1016/bs.mcb.2018.03.026. Epub 2018 Apr 11.
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本文引用的文献

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14-3-3 regulation of Ncd reveals a new mechanism for targeting proteins to the spindle in oocytes.14-3-3对Ncd的调控揭示了一种将蛋白质靶向卵母细胞纺锤体的新机制。
J Cell Biol. 2017 Oct 2;216(10):3029-3039. doi: 10.1083/jcb.201704120. Epub 2017 Aug 31.
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The chromosomal basis of meiotic acentrosomal spindle assembly and function in oocytes.卵母细胞减数分裂无中心体纺锤体组装和功能的染色体基础。
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Kdm5/Lid Regulates Chromosome Architecture in Meiotic Prophase I Independently of Its Histone Demethylase Activity.Kdm5/Lid在减数分裂前期I中独立于其组蛋白去甲基化酶活性调控染色体结构。
PLoS Genet. 2016 Aug 5;12(8):e1006241. doi: 10.1371/journal.pgen.1006241. eCollection 2016 Aug.
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The microtubule catastrophe promoter Sentin delays stable kinetochore-microtubule attachment in oocytes.微管解聚促进因子Sentin延迟卵母细胞中动粒微管的稳定附着。
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Meiosis: an overview of key differences from mitosis.减数分裂:与有丝分裂关键差异的概述
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Meiosis-specific stable binding of augmin to acentrosomal spindle poles promotes biased microtubule assembly in oocytes.减数分裂特异性 augmin 与无中心体纺锤极的稳定结合促进了卵母细胞中偏心的微管组装。
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Depleting gene activities in early Drosophila embryos with the "maternal-Gal4-shRNA" system.利用“母源-Gal4-shRNA”系统在早期果蝇胚胎中消耗基因活性。
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8
The chromosomal passenger complex is required for meiotic acentrosomal spindle assembly and chromosome biorientation.染色体乘客复合物对于减数分裂无着丝粒纺锤体的组装和染色体的定向排列是必需的。
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The conserved kinase SRPK regulates karyosome formation and spindle microtubule assembly in Drosophila oocytes.保守激酶 SRPK 调节果蝇卵母细胞中的核体形成和纺锤体微管组装。
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A genome-scale shRNA resource for transgenic RNAi in Drosophila.用于果蝇中转基因 RNAi 的全基因组规模 shRNA 资源。
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结合显微镜技术和生物化学方法研究果蝇卵母细胞减数分裂纺锤体组装

Combining microscopy and biochemistry to study meiotic spindle assembly in Drosophila oocytes.

作者信息

Romé Pierre, Ohkura Hiroyuki

机构信息

Wellcome Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.

Wellcome Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.

出版信息

Methods Cell Biol. 2018;145:237-248. doi: 10.1016/bs.mcb.2018.03.026. Epub 2018 Apr 11.

DOI:10.1016/bs.mcb.2018.03.026
PMID:29957206
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6031290/
Abstract

Studies using Drosophila have played pivotal roles in advancing our understanding of molecular mechanisms of mitosis throughout the past decades, due to the short generation time and advanced genetic research of this organism. Drosophila is also an excellent model to study female meiosis in oocytes. Pathways such as the acentrosomal assembly of the meiotic spindle in oocytes are conserved from fly to humans. Collecting and manipulating large Drosophila oocytes for microscopy and biochemistry are both time and cost efficient, offering advantages over mouse or human oocytes. Therefore, Drosophila oocytes serve as an excellent platform for molecular studies of female meiosis using a combination of genetics, microscopy, and biochemistry. Here we describe key methods to observe the formation of the meiotic spindle either in fixed or in live oocytes. Moreover, biochemical methods are described to identify protein-protein interactions in vivo.

摘要

在过去几十年里,由于果蝇的世代周期短且遗传研究先进,利用果蝇进行的研究在推动我们对有丝分裂分子机制的理解方面发挥了关键作用。果蝇也是研究卵母细胞中雌性减数分裂的优秀模型。卵母细胞中减数分裂纺锤体的无中心体组装等途径在从果蝇到人类中都是保守的。收集和操作大量果蝇卵母细胞用于显微镜检查和生物化学研究既节省时间又成本效益高,比小鼠或人类卵母细胞具有优势。因此,果蝇卵母细胞是使用遗传学、显微镜检查和生物化学相结合的方法对雌性减数分裂进行分子研究的优秀平台。在这里,我们描述了在固定或活卵母细胞中观察减数分裂纺锤体形成的关键方法。此外,还描述了用于鉴定体内蛋白质 - 蛋白质相互作用的生物化学方法。