Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama 226-8503, Japan.
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan.
Biol Open. 2023 May 15;12(5). doi: 10.1242/bio.059783. Epub 2023 May 17.
In the cytoplasm, filamentous actin (F-actin) plays a critical role in cell regulation, including cell migration, stress fiber formation, and cytokinesis. Recent studies have shown that actin filaments that form in the nucleus are associated with diverse functions. Here, using live imaging of an F-actin-specific probe, superfolder GFP-tagged utrophin (UtrCH-sfGFP), we demonstrated the dynamics of nuclear actin in zebrafish (Danio rerio) embryos. In early zebrafish embryos up to around the high stage, UtrCH-sfGFP increasingly accumulated in nuclei during the interphase and reached a peak during the prophase. After nuclear envelope breakdown (NEBD), patches of UtrCH-sfGFP remained in the vicinity of condensing chromosomes during the prometaphase to metaphase. When zygotic transcription was inhibited by injecting α-amanitin, the nuclear accumulation of UtrCH-sfGFP was still observed at the sphere and dome stages, suggesting that zygotic transcription may induce a decrease in nuclear F-actin. The accumulation of F-actin in nuclei may contribute to proper mitotic progression of large cells with rapid cell cycles in zebrafish early embryos, by assisting in NEBD, chromosome congression, and/or spindle assembly.
在细胞质中,丝状肌动蛋白(F-actin)在细胞调节中起着关键作用,包括细胞迁移、应力纤维形成和胞质分裂。最近的研究表明,形成在核中的肌动蛋白丝与多种功能相关。在这里,我们使用 F-actin 特异性探针的活细胞成像,超折叠 GFP 标记的 utrophin(UtrCH-sfGFP),在斑马鱼(Danio rerio)胚胎中展示了核中肌动蛋白的动态。在早期斑马鱼胚胎中,直到高期,UtrCH-sfGFP 在有丝分裂间期逐渐积累在核中,并在前期达到峰值。核膜破裂(NEBD)后,在核膜破裂后,UtrCH-sfGFP 的斑点仍然存在于浓缩染色体附近,处于前中期到中期。当用α-鹅膏蕈碱注射抑制合子转录时,在球期和穹顶期仍观察到 UtrCH-sfGFP 的核积累,这表明合子转录可能导致核 F-actin 减少。F-actin 在核中的积累可能有助于斑马鱼早期胚胎中大细胞的有丝分裂进程,通过协助核膜破裂、染色体聚集和/或纺锤体组装。