Department of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital / West China School of Medicine, Sichuan University.
Department of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital / West China School of Medicine, Sichuan University;
J Vis Exp. 2024 Jul 26(209). doi: 10.3791/66458.
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.
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（Danio rerio）已被广泛用作研究从卵母细胞到胚胎的早期发育过程的模式生物。在斑马鱼中，卵母细胞被一层单独的体腔颗粒细胞包围。然而，分离颗粒细胞和卵母细胞是一个挑战，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不同发育阶段的斑马鱼卵母细胞，但目前的技术在完全去除颗粒细胞方面存在不足，限制了仅针对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，可以在斑马鱼中分离出纯的 I 期卵母细胞，同时消除颗粒细胞的污染。该技术促进了生化和分子分析，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，该方法用户友好，最小化了卵母细胞的损伤，为后续的研究和分析提供了实用的解决方案。
