University Center of Tacuarembó,UDELAR, Route 5, Km 386, Tacuarembó,Uruguay.
National Agricultural Research Institute,INIA, Route 50, Km 11, Colonia 70000,Uruguay.
Microsc Microanal. 2019 Aug;25(4):1032-1036. doi: 10.1017/S1431927619000692. Epub 2019 May 28.
The aims of the present study were to: (i) evaluate the ultrastructural differences in the zona pellucida (ZP) surface between immature and mature bovine oocytes, and (ii) describe a new objective technique to measure the pores in the outer ZP. Intact cumulus-oocyte complexes (COCs) obtained from a local abattoir were immediately fixed (immature group) or submitted to in vitro maturation (IVM) at 38.5 °C for 24 h in a humidified atmosphere of 5% CO2 in air (mature group). Oocytes from both groups were morphologically evaluated via Scanning Electron Microscopy (SEM) and the images were processed in the Fiji/ImageJ software using a new objective methodology through the Trainable Weka Segmentation plugin. The average number of pores in ZP was greater (p 0.05) between groups. In conclusion, it has been shown that the number of pores highlighted the main ultrastructural change in the morphology of the ZP surface of bovine oocytes during the IVM process. We have described an objective method that can be used to evaluate ultrastructural modifications of the ZP surface during oocyte maturation and early embryo development.
（i）评估不成熟和成熟牛卵母细胞透明带（ZP）表面的超微结构差异，（ii）描述一种新的客观技术来测量 ZP 外孔。从当地屠宰场获得的完整卵丘-卵母细胞复合物（COCs）立即固定（不成熟组）或在 38.5°C 下在 5%CO2 加湿气氛中进行体外成熟（IVM）24 小时（成熟组）。通过扫描电子显微镜（SEM）对两组卵母细胞进行形态评估，并使用新的客观方法通过 Trainable Weka Segmentation 插件在 Fiji/ImageJ 软件中处理图像。ZP 中的孔的平均数量更大（p 0.05）。总之，已经表明，在 IVM 过程中，ZP 表面形态的主要超微结构变化突出了孔的数量。我们已经描述了一种客观的方法，可用于评估卵母细胞成熟和早期胚胎发育过程中 ZP 表面的超微结构变化。
