利用卵巢荧光转基因斑马鱼实时观察卵母细胞减数分裂过程中胚芽囊泡的迁移。

Zebrafish Pub Date : 2024-04-01 DOI:10.1089/zeb.2023.0048
Eisei Tsutsumi, Toshinobu Tokumoto
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引用次数: 0

摘要

通过转基因(TG)斑马鱼,研究人员可以对体内细胞和组织中的特定生物现象进行生物成像。我们建立 TG 品系是为了监测活鱼卵巢的变化。最初的 TG 品系偶尔会出现卵巢荧光。虽然该品系中整合的 cDNA 是由青鳉 β-肌动蛋白启动子驱动的增强型绿色荧光蛋白(EGFP)的表达,但 EGFP 的表达仅限于成鱼的卵母细胞和鳃。此外,我们还发现,已建立品系的卵母细胞中的生殖泡(GV)可通过 GV 周围相对较强的荧光观察到。在这项研究中,我们试图利用 GV 荧光卵母细胞捕捉减数分裂细胞分裂过程中生殖泡破裂(GVBD)的动态过程。因此,GV迁移和GVBD可被实时监测。我们还成功地观察到了参与 GV 向动物极迁移的肌动蛋白丝。该菌株可用于卵母细胞减数分裂过程的教育。
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Real-Time Observation of Germinal Vesicle Migration During Oocyte Meiotic Cell Division Using Ovarian Fluorescent Transgenic Zebrafish.
The transgenic (TG) zebrafish allows researchers to bio-image specific biological phenomena in cells and tissues in vivo. We established TG lines to monitor changes in the ovaries of live fish. The original TG line with ovarian fluorescence was occasionally established. Although the cDNA integrated into the line was constructed for the expression of enhanced green fluorescent protein (EGFP) driven by the medaka β-actin promoter, the expression of EGFP is restricted to the oocytes and gills in adult fish. Furthermore, we found that germinal vesicles (GVs) in oocytes of the established line can be observed by relatively strong fluorescence around the GV. In this study, we tried to capture the dynamic processes of germinal vesicle breakdown (GVBD) during meiotic cell division using the GV fluorescent oocytes. As a result, GV migration and GVBD could be monitored in real time. We also succeeded in observing actin filaments involved in the migration of GV to the animal pole. This strain can be used for education in the process of oocyte meiotic cell division.
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