缝隙连接蛋白43和黄体生成素受体在小鼠卵巢中的表达

doi:10.16098/j.issn.0529-1356.2017.03.016

解剖学报 ›› 2017, Vol. 48 ›› Issue (3): 342-347.doi: 10.16098/j.issn.0529-1356.2017.03.016

• 组织学胚胎学发育生物学 • 上一篇下一篇

缝隙连接蛋白43和黄体生成素受体在小鼠卵巢中的表达

经华高山庆黑常春蔡玉芳孔斌郑小敏吴凯赵承军* 常青*

宁夏医科大学基础医学院人体解剖学与组织学胚胎学系，生育力保持教育部重点实验室，宁夏生殖与遗传重点实验室，银川 750004

收稿日期:2016-09-26 修回日期:2016-11-18 出版日期:2017-06-06 发布日期:2017-09-19
通讯作者: 赵承军;常青 E-mail:seryin55@163.com
基金资助:
黄体生成素对多囊卵巢综合症卵巢缝隙连接蛋白表达调节的研究、促卵泡激素对睾丸支持细胞胶质细胞源性神经营养因子表达调节的机理研究、枸杞多糖干预对提高小鼠冻存卵巢移植效果的相关机制研究

Expression of connexin 43 and luteinizing hormone receptor in mouse ovary

JING Hua GAO Shan-qing HEI Chang-chun CAI Yu-fang KONG Bin ZHENG Xiao-min WU Kai ZHAO Cheng-jun* CHANG Qing*

Department of Anatomy, Histology and Embryology, School of Basic Medicine, Ningxia Medical University; Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education; Key Laboratory of Reproduction and Genetics in Ningxia, Yinchuan 750004, China

Received:2016-09-26 Revised:2016-11-18 Online:2017-06-06 Published:2017-09-19
Contact: ZHAO Cheng-jun; CHANG Qing* E-mail:seryin55@163.com

摘要/Abstract

摘要：

目的观察小鼠卵泡发育过程中缝隙连接蛋白43（Cx43）和黄体生成素受体（LHR）在小鼠卵巢中的表达以及黄体生成素(LH)对卵巢Cx43表达的影响，为这两种分子与卵泡发育的关系提供实验依据。方法选择出生后1d、4d、7d、2周、3周、4周、6周、8周龄雌性C57小鼠共8组，每组10只，取卵巢。HE染色观察卵巢形态、卵泡发育并测量颗粒细胞体积分数。免疫组织化学和Western blotting观察卵巢组织中Cx43及LHR的表达，Western blotting检测不同浓度黄体生成素（LH）孵育后对卵巢Cx43表达的影响。结果 HE染色显示，出生1d小鼠卵巢中见原始卵泡，出生7d卵巢内出现初级卵泡，2周时出现窦前卵泡和少量窦卵泡，3周、4周时，出现较多大的窦卵泡，6～8周时出现黄体。卵泡颗粒细胞体积分数在2周后的卵巢中明显增加。Cx43表达在各阶段卵泡的颗粒细胞胞膜上，LHR表达在卵泡膜细胞、颗粒细胞及卵母细胞胞质中。Cx43和LHR出生后2周卵巢中的表达开始明显增加，并随卵泡发育表达逐渐增强，在3周、4周、6周和8周维持相对较高水平。LH体外干预可增加卵巢组织Cx43的表达。结论 Cx43和LHR在卵泡发育中发挥重要作用，LH可能通过LHR促进卵巢卵泡颗粒细胞Cx43的表达。

关键词: 缝隙连接蛋白43, 黄体生成素受体, 颗粒细胞, 小鼠, 免疫组织化学, 免疫印迹法, 小鼠

Abstract:

Objective To observe the expression of connexin 43 (Cx43) and luteinizing hormone receptor(LHR) during the follicular development in mouse ovaries and the effect of luteinizing hormone(LH) on the expression of Cx43, providing experimental evidence for the relationship between the two molecules and the follicular development. Methods Eighty mice were divided into 8 groups with the ages of 1-day-old, 4-day-old, 7-day-old, 2-week-old, 3-week-old, 4-week-old, 6-week-old and 8-week-old, 10 mice for each group. Hematoxylineosin (HE) staining was used to observe the ovarian morphology and calculated the volume fraction of granulosa cells. Immunohistochemistry and Western blotting were used to observe the expression of Cx43 and LHR in ovarian tissues. Western blotting was used to observe the expression of Cx43 in the ovaries after incubation with different concentrations of LH. Results The HE staining results showed that there were primordial follicles, primary follicles in 1-day-old, 7-day-old, 2-week-old and 3 to 4-week-old mouse ovaries, respectively, corpus luteum appeared in 6 to 8-week-old mouse ovaries. The volume fraction of granulosa cells was significantly increased in the ovary after 2-week-old. The results of immunohistochemistry showed that Cx43 expressed in cell membrane of granulosa cells at different stages of the follicles and LHR was expressed in the cytoplasm of the theca cells, granulosa cells and oocytes. After 2 weeks of birth, the expression of Cx43 and LHR increased significantly in ovaries, and raised gradually with the follicular development and then maintained a relatively high level at 3-week-old, 4-week-old, 6-week-old and 8-week-old mouse ovaries. Intervention of LH increased the expression of Cx43 in ovarian tissues. Conclusion Cx43 and LHR play important roles in follicular development and the expression of Cx43 in granulosa cells may be regulated by LH through LHR.

Key words: Connexin 43, Luteinizing hormone receptor, Granulosa cells, Mouse, Immunohistochemistry, Western blotting, Mouse

经华高山庆黑常春蔡玉芳孔斌郑小敏吴凯赵承军常青. 缝隙连接蛋白43和黄体生成素受体在小鼠卵巢中的表达[J]. 解剖学报, 2017, 48(3): 342-347.

JING Hua GAO Shan-qing HEI Chang-chun CAI Yu-fang KONG Bin ZHENG Xiao-min WU Kai ZHAO Cheng-jun CHANG Qing. Expression of connexin 43 and luteinizing hormone receptor in mouse ovary[J]. Acta Anatomica Sinica, 2017, 48(3): 342-347.

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缝隙连接蛋白43和黄体生成素受体在小鼠卵巢中的表达

Expression of connexin 43 and luteinizing hormone receptor in mouse ovary

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