貉卵母细胞成熟发育的细胞学研究

doi:10.16431/j.cnki.1671-7236.2023.04.015

摘要/Abstract

摘要： 【目的】探究貉卵母细胞成熟发育特征,为貉卵母细胞的体外成熟培养及优质成熟卵母细胞的挑选提供依据,以提高貉的体外受精效率。【方法】采集1~3岁(性成熟)繁殖期母貉的新鲜卵巢,制备光、电镜样品,观察貉卵母细胞成熟发育过程中细胞器变化;利用X射线微分析技术探索卵母细胞质膜化学元素变化与成熟发育的关系。【结果】根据颗粒细胞层数、卵泡直径、卵母细胞大小以及透明带的变化,将貉卵母细胞成熟发育划分为8个时期。貉卵母细胞在早期发育阶段(第Ⅰ~Ⅳ期)由颗粒细胞包围,逐渐出现透明带、卵泡腔,生发泡内的染色质越来越致密;随着卵母细胞的成熟发育(第Ⅴ~Ⅶ期),微绒毛、皮质颗粒出现;高尔基复合体、线粒体等细胞器数量明显增加,且逐渐迁移至皮质区,同时线粒体、脂滴形态发生变化。卵母细胞成熟时(第Ⅷ期),高尔基复合体及粗面内质网消失,皮质颗粒在质膜下排列;排卵后,核仁发生致密化。随着卵泡腔的增大,卵母细胞质膜表面的钠、氯和硫含量呈下降趋势,第Ⅴ期卵母细胞质膜中钠、氯和硫含量显著高于第Ⅶ期(P＜0.05),钙含量显著低于第Ⅵ期和第Ⅶ期卵母细胞(P＜0.05);钾含量呈现低-高-低的变化规律,第Ⅵ期卵母细胞质膜中钾含量显著高于第Ⅴ期和第Ⅵ期(P＜0.05)。【结论】貉卵母细胞发育至第Ⅴ期后细胞器的种类和数量增多,质膜化学元素含量发生变化,卵母细胞膜电位发生去极化,开始进入成熟发育阶段,此时的卵母细胞具备体外发育成熟的能力。

关键词: 貉卵母细胞; 成熟发育; 细胞器; 化学元素

Abstract: 【Objective】 The purpose of this study was to explore the maturation characteristics of raccoon dog oocytes,and to provide a basis for the in vitro maturation culture of raccoon dog oocytes and to select high-quality mature oocytes,so as to improve the in vitro fertilization efficiency of raccoon dog.【Method】 Fresh ovaries from 1 to 3 years old female raccoon dogs at reproductive stage were collected to prepare light and electron microscope samples to observe the changes of organelles during oocyte maturation of raccoon dog. X-ray microanalysis was used to explore the relationship between the changes of chemical elements in plasma membrane and maturation of oocyte.【Result】 The maturation and development of raccoon dog oocytes could be divided into 8 stages according to the layer number of granulosa cells,the diameter of follicles,the size of oocytes and the changes of the zona pellucida.Raccoon dog oocytes were surrounded by granulosa cells in the early developmental stage (stages Ⅰ to Ⅳ),with the gradual appearance of the zona pellucida and the follicular cavity,the chromatin in the germinal vesicles became more and more compact.With the maturation and development of oocytes (stages Ⅴ to Ⅶ),cortical granules and microvilli appeared and the number of organelles such as Golgi complex,mitochondria increased significantly and they gradually migrated to the cortex,while mitochondria and lipid droplets experienced morphologic variation.When the oocyte grew mature (stage Ⅷ),the Golgi complex and the rough endoplasmic reticulum disappeared,and the cortical granules were arranged under the plasmalemma.After ovulation,the nucleolus became compact.The contents of sodium,chloride and sulfur in oocyte plasma membrane decreased with the increase of follicular cavity.The contents of sodium,chlorine and sulfur in oocyte plasma membrane in stage Ⅴ were significantly higher than those in stage Ⅶ (P＜0.05),and the relative content of calcium was significantly lower than that of stage Ⅵ and Ⅶ oocytes (P＜0.05).Potassium showed a low-high-low variation,and the content of potassium in oocyte plasma membrane in stage Ⅵ was significantly higher than that of stage Ⅴ and Ⅵ oocytes (P＜0.05).【Conclusion】 After the raccoon dog oocytes developed to stage Ⅴ,the type and number of organelles increased,the plasma membrane chemical elements changed,and oocyte membrane potential was depolarized,and the oocytes began to enter the mature stage.At this time,the oocytes had the ability to mature in vitro.

Key words: raccoon dog oocyte; maturation development; organelles; chemical elements

中图分类号:

S865.22

唐毓, 杨镒峰, 张颖, 薛海龙, 冯怀亮, 许保增. 貉卵母细胞成熟发育的细胞学研究[J]. 中国畜牧兽医, 2023, 50(4): 1434-1443.

TANG Yu, YANG Yifeng, ZHANG Ying, XUE Hailong, FENG Huailiang, XU Baozeng. Cytological Study on Oocyte Maturation of Raccoon Dog[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1434-1443.

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