Abstract:
The study of oocyte development holds significant implications in developmental biology. The zebrafish (Danio rerio) has been extensively used as a model organism to investigate early developmental processes from oocyte to embryo. In zebrafish, oocytes are surrounded by a single layer of somatic granulosa cells. However, separating granulosa cells from oocytes poses a challenge, as achieving pure oocytes is crucial for precise analysis. Although various methods have been proposed to isolate zebrafish oocytes at different developmental stages, current techniques fall short in removing granulosa cells completely, limiting the accuracy of genome analysis focused solely on oocytes. In this study, we successfully developed a rapid and efficient process for isolating pure stage I oocytes in zebrafish while eliminating granulosa cell contamination. This technique facilitates biochemical and molecular analysis, particularly in exploring epigenetic and genome structure aspects specific to oocytes. Notably, the method is user-friendly, minimizes oocyte damage, and provides a practical solution for subsequent research and analysis.
摘要:
卵母细胞发育的研究在发育生物学中具有重要意义。斑马鱼(Daniorerio)已被广泛用作模型生物,以研究从卵母细胞到胚胎的早期发育过程。在斑马鱼中,卵母细胞被单层体细胞颗粒细胞包围。然而,从卵母细胞中分离颗粒细胞是一个挑战,因为获得纯卵母细胞对于精确分析至关重要。尽管已经提出了各种方法来分离处于不同发育阶段的斑马鱼卵母细胞,目前的技术不足以完全去除颗粒细胞,限制了只关注卵母细胞的基因组分析的准确性.在这项研究中,我们成功开发了一种快速有效的方法,用于分离斑马鱼中的纯I期卵母细胞,同时消除颗粒细胞污染。这种技术有助于生化和分子分析,特别是在探索卵母细胞特有的表观遗传和基因组结构方面。值得注意的是,该方法是用户友好的,最大限度地减少卵母细胞的损伤,为后续研究和分析提供了切实可行的解决方案。