华北农学报 ›› 2023, Vol. 38 ›› Issue (2): 217-223. doi: 10.7668/hbnxb.20193551

所属专题: 畜牧 生物技术

• 畜牧·水产·兽医 • 上一篇    下一篇

小尾寒羊BMF基因在发情不同阶段卵巢及卵泡发育过程中的表达分析

李营营, 张昊堃, 贾皓帆, 郭婧瑶, 杨喜喜, 王志亮, 李明娜, 王欣荣   

  1. 甘肃农业大学 动物科学技术学院,甘肃 兰州 730070
  • 收稿日期:2022-12-07 出版日期:2023-04-28
  • 通讯作者:
    李明娜(1987-),女,甘肃景泰人,讲师,博士,主要从事动物繁殖原理与技术应用研究。
    王欣荣(1974-),男,甘肃临潭人,教授,博士,硕士生导师,主要从事动物繁殖原理与技术应用研究。
  • 作者简介:

    李营营(1996-),女,甘肃静宁人,在读硕士,主要从事动物繁殖原理与技术应用研究。

  • 基金资助:
    国家自然科学基金(31960664); 国家自然科学基金(31560634); 甘肃省自然科学基金(21JR7RA809); 甘肃农业大学人才专项(2017RCZX-17); 甘肃农业大学学科团队项目(GAU-XKTD-2022-21)

Expression Analysis of BMF Gene During the Ovaries at Different Stages of Estrus and Follicular Development in Small-tailed Han Sheep

LI Yingying, ZHANG Haokun, JIA Haofan, GUO Jingyao, YANG Xixi, WANG Zhiliang, LI Mingna, WANG Xinrong   

  1. College of Animal Science and Technology,Gansu Agricultural University, Lanzhou 730070,China
  • Received:2022-12-07 Published:2023-04-28

摘要:

旨在初探BMF基因在小尾寒羊卵泡发育过程中的生物学功能,为将来深入探索其调控机理提供理论依据。对小尾寒羊实施同期发情,采用实时荧光定量PCR(Quantitative real-time PCR,RT-qPCR)、蛋白免疫印迹(Western Blot,WB)及免疫荧光染色(IF)方法检测BMF基因及其编码蛋白在不同生理阶段卵巢中的表达量;随后分离不同直径的卵泡,分析目的基因在大、中、小卵泡中的表达差异。结果显示,BMF mRNA在撤栓后42 h卵巢中的表达量显著高于撤栓后18 h(P<0.05),其编码蛋白表达结果与mRNA基本一致;BMF蛋白主要表达于卵泡颗粒细胞和卵泡膜细胞中;进一步分离卵泡,发现BMF基因及其编码蛋白表达量均在中卵泡中显著高于大卵泡和小卵泡(P<0.05)。以上结果表明,BMF基因可能参与卵巢周期性活动,在卵泡发育过程中发挥重要作用。

关键词: 小尾寒羊, BMF, 卵泡, 卵巢, 繁殖

Abstract:

The aim of this experiment was to investigate the biological function of BMF gene(Bcl-2 modifying factor)in the follicular development of Small-tailed Han Sheep,and to provide the theoretical basis for further exploration of its regulatory mechanism in the future.Firstly,the synchronization estrus was implemented in Small-tailed Han Sheep.Then,we used Quantitative Real-time PCR(RT-qPCR),Western Blot(WB)and Immunofluorescence staining(IF)to detect the expression of the BMF gene and its encoded protein in the ovaries at different physiological stages;subsequently,we isolated follicles of different diameters and analyzed the differences in expression of the target gene in large,middle and small follicles.The results showed that the expression of BMF mRNA was significantly higher in the ovary at 42 h after thrombectomy than at 18 h after thrombectomy(P<0.05),and the results of its encoded protein expression of BMF were basically consistent with the mRNA.BMF protein was mainly expressed in follicular granulosa cells and follicular membrane cells.We further isolated follicles and found that the expression of both the BMF gene and its encoded protein in middle follicles was significantly higher than that in large and small follicles(P<0.05).These results suggest that BMF gene may be involved in the ovarian cyclic activity and play an important role in follicular development.

Key words: Small-tailed Han Sheep, BMF, Follicle, Ovary, Reproduction

引用本文

李营营, 张昊堃, 贾皓帆, 郭婧瑶, 杨喜喜, 王志亮, 李明娜, 王欣荣. 小尾寒羊BMF基因在发情不同阶段卵巢及卵泡发育过程中的表达分析[J]. 华北农学报, 2023, 38(2): 217-223. doi: 10.7668/hbnxb.20193551.

LI Yingying, ZHANG Haokun, JIA Haofan, GUO Jingyao, YANG Xixi, WANG Zhiliang, LI Mingna, WANG Xinrong. Expression Analysis of BMF Gene During the Ovaries at Different Stages of Estrus and Follicular Development in Small-tailed Han Sheep[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(2): 217-223. doi: 10.7668/hbnxb.20193551.

使用本文

0
    /   /   推荐 /   导出引用

链接本文: http://www.hbnxb.net/CN/10.7668/hbnxb.20193551

               http://www.hbnxb.net/CN/Y2023/V38/I2/217