Wellcome Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3BF, UK.
Chromosoma. 2023 Mar;132(1):1-18. doi: 10.1007/s00412-023-00784-9. Epub 2023 Jan 17.
In prophase of the first meiotic division, chromatin forms a compact spherical cluster called the karyosome within the enlarged oocyte nucleus in Drosophila melanogaster. Similar clustering of chromatin has been widely observed in oocytes in many species including humans. It was previously shown that the proper karyosome formation is required for faithful chromosome segregation, but knowledge about its formation and maintenance is limited. To identify genes involved in karyosome formation, we carried out a large-scale cytological screen using Drosophila melanogaster oocytes. This screen comprised 3916 genes expressed in ovaries, of which 106 genes triggered reproducible karyosome defects upon knockdown. The karyosome defects in 24 out of these 106 genes resulted from activation of the meiotic recombination checkpoint, suggesting possible roles in DNA repair or piRNA processing. The other genes identified in this screen include genes with functions linked to chromatin, nuclear envelope, and actin. We also found that silencing of genes with mitochondrial functions, including electron transport chain components, induced a distinct karyosome defect typically with de-clustered chromosomes located close to the nuclear envelope. Furthermore, mitochondrial dysfunction not only impairs karyosome formation and maintenance, but also delays synaptonemal complex disassembly in cells not destined to become the oocyte. These karyosome defects do not appear to be mediated by apoptosis. This large-scale unbiased study uncovered a set of genes required for karyosome formation and revealed a new link between mitochondrial dysfunction and chromatin organization in oocytes.
在第一次减数分裂前期,染色质在果蝇的扩大卵细胞核内形成一个称为核体的紧密的球形簇。在包括人类在内的许多物种的卵母细胞中,广泛观察到类似的染色质聚集。先前的研究表明,核体的正确形成对于染色体的正确分离是必需的,但对其形成和维持的了解有限。为了鉴定参与核体形成的基因,我们利用果蝇卵母细胞进行了大规模的细胞学筛选。该筛选包括在卵巢中表达的 3916 个基因,其中 106 个基因在敲低后会导致核体缺陷的可重复出现。这 106 个基因中的 24 个核体缺陷是由于减数分裂重组检查点的激活引起的,这表明它们可能在 DNA 修复或 piRNA 处理中发挥作用。在这个筛选中鉴定的其他基因包括与染色质、核膜和肌动蛋白相关的功能的基因。我们还发现,沉默具有线粒体功能的基因,包括电子传递链组件,会诱导一种独特的核体缺陷,通常是染色体去聚集,位于核膜附近。此外,线粒体功能障碍不仅会损害核体的形成和维持,还会延迟注定不成为卵母细胞的细胞中联会复合体的解体。这些核体缺陷似乎不是由细胞凋亡介导的。这项大规模的无偏研究揭示了一组形成核体所必需的基因,并揭示了线粒体功能障碍与卵母细胞中染色质组织之间的新联系。