白血病抑制因子对牛卵母细胞体外发育能力的影响

doi:10.16431/j.cnki.1671-7236.2024.02.003

中国畜牧兽医 ›› 2024, Vol. 51 ›› Issue (2): 462-469.doi: 10.16431/j.cnki.1671-7236.2024.02.003

白血病抑制因子对牛卵母细胞体外发育能力的影响

莫显红^1,4, 陶庆华², 房义³, 梁婧¹, 张佳音¹, 梁小航¹, 徐振军¹

1. 赤峰学院化学与生命科学学院, 赤峰 024000;
2. 福建农林大学动物科学学院(蜂学学院), 福州 350002;
3. 中国科学院东北地理与农业生态研究所吉林省草地畜牧重点实验室, 长春 130062;
4. 内蒙古赛科星家畜种业与繁育生物技术研究院, 呼和浩特 011517

收稿日期:2023-10-07 出版日期:2024-02-05 发布日期:2024-01-29
作者简介:莫显红,E-mail:15652502953@163.com。

Effect of Leukemia Inhibitory Factor on Developmental Capacity of Bovine Oocyte

MO Xianhong^1,4, TAO Qinghua², FANG Yi³, LIANG Jing¹, ZHANG Jiayin¹, LIANG Xiaohang¹, XU Zhenjun¹

1. College of Chemistry and Life Science, Chifeng University, Chifeng 024000, China;
2. College of Animal Science (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, China;
3. Jilin Provincial Key Laboratory of Grassland Farming, Northeast Institute of Geography and Agoecology, Chinese Academy of Sciences, Changchun 130062, China;
4. Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Hohhot 011517, China

Received:2023-10-07 Online:2024-02-05 Published:2024-01-29
Contact: 内蒙古自治区自然科学基金(2022LHMS03010);内蒙古自治区高等学校科学研究项目(NJZY22130);赤峰学院大学生创新创业训练计划项目(S202310138024);赤峰学院一流学科建设项目(CFXYYLXKB202102);蒙东地区生物资源开发利用创新团队(cfxykycxtd202008) E-mail:linkur3@126.com
Supported by:
The project was supported by the National Key Research and Development Program of China (2019YFC1905301);National Natural Science Foundation of China (22078115,21776108,21690083,22008078).

摘要/Abstract

摘要： 【目的】探究白血病抑制因子(leukemia inhibitory factor,LIF)对牛卵母细胞和早期胚胎发育能力及多潜能基因表达的影响。【方法】将牛卵丘-卵母细胞复合体(cumulus oocyte complexes,COCs)分为4组,对照组:在卵母细胞体外成熟(in vitro maturation,IVM)和胚胎体外培养(in vitro culture,IVC)过程中均不添加LIF溶液;处理组1(T1组):IVM过程添加25 ng/mL LIF,IVC过程不添加LIF;处理组2(T2组):IVM过程不添加LIF,IVC过程添加25 ng/mL LIF;处理组3(T3组):IVM和IVC全过程均添加25 ng/mL LIF。用Fluo-3荧光指示剂检测卵母细胞胞内游离钙离子([Ca²⁺]i)浓度;挑选有第一极体排出的卵母细胞进行孤雌激活,统计各组卵母细胞卵裂率和囊胚率;利用实时荧光定量PCR检测囊胚中多潜能基因的相对表达量。【结果】与对照组相比,培养8 h后,T1组牛卵母细胞胞内[Ca²⁺]i浓度极显著下降(P＜0.01);培养24 h后,T1组牛卵母细胞胞内[Ca²⁺]i浓度极显著升高(P＜0.01)。T1和T3组孤雌激活后卵裂率和囊胚率均显著高于对照组,且T3组囊胚率显著高于T1和T2组(P＜0.05)。实时荧光定量PCR结果显示,T3组囊胚中POU结构域5类转录因子1(POU5F1)和同源框蛋白(NANOG)基因表达量均显著高于对照组,尾型同源盒转录因子2(CDX2)基因表达量显著低于对照组(P＜0.05)。【结论】在IVM过程中添加LIF可提高卵母细胞的发育能力,在IVM＋IVC全过程添加LIF对早期胚胎发育和多潜能基因的表达有积极作用。

关键词: 白血病抑制因子; 多潜能基因; 钙离子; 胚胎发育

Abstract: 【Objective】 This experiment was to explore the effect of leukemia inhibitory factor (LIF) on the development ability of bovine oocytes and early embryo and the expression of multipotent genes.【Method】 The cumulus-oocyte-complexes (COCs) of bovine were divided into 4 groups.In the control group, no LIF solution was added during in vitro maturation (IVM) and embryo in vitro culture (IVC).In treatment group 1 (T1 group), 25 ng/mL LIF was added to the IVM process and no LIF was added to the IVC process.In treatment group 2 (T2 group), the IVM process did not add LIF, and the IVC process added 25 ng/mL LIF.In treatment group 3 (T3 group), 25 ng/mL LIF was added to the whole process of IVM and IVC.The concentration of intracellular free calcium ([Ca²⁺]i) in oocytes was detected by Fluo-3 fluorescence indicator.Oocytes with the first polar body were selected for parthenogenetic activation, and the cleavage rate and blastocyst rate of each group were calculated.Real-time quantitative PCR was used to detect the relative expression of multipotent genes.【Result】 Compared with the control group, intracellular [Ca²⁺]i concentration of bovine oocytes in T1 group was extremely significantly decreased after 8 h of cultivation (P<0.01).After 24 h of cultivation, the intracellular [Ca²⁺]i concentration of bovine oocytes in T1 group was extremely significantly increased (P<0.01).The cleavage rate and blastocyst rate in T1 and T3 groups were significantly higher than those in control group, and the blastocyst rate in T3 group was significantly higher than that in T1 and T2 groups (P<0.05).The results of Real-time quantitative PCR showed that the expression levels of POU5F1 and NANOG genes in T3 group were significantly higher than those in control group, and the expression level of CDX2 gene was significantly lower than that in control group (P<0.05).【Conclusion】 The addition of LIF during IVM could improve the developmental ability of oocytes, and the addition of LIF during the whole process of IVM＋IVC had a positive effect on early embryonic development and the expression of multipotent genes.

Key words: leukemia inhibitory factor; multipotent gene; calcium ions; embryonic development

中图分类号:

S852.2

莫显红, 陶庆华, 房义, 梁婧, 张佳音, 梁小航, 徐振军. 白血病抑制因子对牛卵母细胞体外发育能力的影响[J]. 中国畜牧兽医, 2024, 51(2): 462-469.

MO Xianhong, TAO Qinghua, FANG Yi, LIANG Jing, ZHANG Jiayin, LIANG Xiaohang, XU Zhenjun. Effect of Leukemia Inhibitory Factor on Developmental Capacity of Bovine Oocyte[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(2): 462-469.

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白血病抑制因子对牛卵母细胞体外发育能力的影响

Effect of Leukemia Inhibitory Factor on Developmental Capacity of Bovine Oocyte

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