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生物钟基因Rev-erb-α缺失导致雌鼠生理周期延长

张芹, 董珍, 吴曦, 颜洁, 高基民

温州医科大学浙江省模式生物技术与应用重点实验室，温州 325035；南京大学模式动物研究所模式动物疾病研究中心MOE重点实验室，南京 210061；苏州大学系统生物学中心，苏州 215006

摘要

在哺乳动物中，生物节律与生殖生理和生殖内分泌密切相关，流行病学研究显示长期倒时差的女性表现出生殖力低下和不良妊娠的概率更高，但是人们对生物节律影响生殖的机制了解还很少。本研究旨在以时钟基因Rev-erb-α敲除型(Rev-erb-α−/−)雌性小鼠为模型初步探索生物节律对生殖的影响。将Rev-erb-α−/− 和野生型(Rev-erb-α+/+)小鼠与C57BL/6J小鼠交配观察生殖力变化，用苏木精-伊红(HE)染色法观察卵巢组织变化，将Rev-erb-α+/+与Rev-erb-α−/−雌性小鼠分别与C57BL/6J 雄性小鼠交配后比较见栓后1.5 d统计的胚胎数量，用阴道涂片结合周期图和周期谱观察小鼠生理周期变化。结果显示，Rev-erb-α−/−雌性小鼠与C57BL/6J雄鼠交配是不生育的，并且Rev-erb-α−/−雌性小鼠生成的胚胎数量明显减少。Rev-erb-α−/−雌性小鼠卵巢存在各级卵泡，并且结构与野生型(Rev-erb-α+/+)雌性小鼠相比无明显差异。Rev-erb-α−/−雌性小鼠的生理周期较Rev-erb-α+/+小鼠周期长约3.22 d，较Rev-erb-α+/+雌性小鼠延长63.5%。以上结果提示，生物钟基因Rev-erb-α缺失导致雌性小鼠生理周期明显延长，可能是影响雌性小鼠生育的原因之一。因此，生物节律的稳定对于生理周期具有重要作用。

关键词： Rev-erb-α; 生理周期; 阴道涂片; 生物节律

分类号：R339.2

[Circadian clock gene Rev-erb-α deficiency prolongs female estrous cyclicity.] [Article in Chinese]

ZHANG Qin, DONG Zhen, WU Xi, YAN Jie, GAO Ji-Min

Zhejiang Province Key Laboratory of biotechnology and application mode, Wenzhou Medical University, Wenzhou 325035, China； MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing University, Nanjing 210061, China； Center for Systems Biology, Soochow University, Suzhou 215006, China

Abstract

The circadian clock has been linked to female reproductive physiology and endocrine in mammals. Epidemiological studies of female shift workers have shown increased rates of abnormal reproduction and adverse pregnancy. But little is known how the circadian rhythms affect reproduction. The aim of the present study was to investigate the influences of circadian rhythms on estrous cycle in female mice using clock gene Rev-erb-α knock out (Rev-erb-α^-/-) mice. To test the fertility of Rev-erb-α^-/- mice, litter sizes were counted after mating with C57BL/6J male mice. HE staining was used to observe the change of follicle development. The number of embryos of Rev-erb-α^+/+ and Rev-erb-α^-/- female mice was compared 1.5 d after mating with C57BL/6J male mice. Then Rev-erb-α^+/+ and Rev-erb-α^-/- female mice were housed to adult, and daily vaginal lavage with 0.9% saline was used to monitor estrous cycle for at least 30 days. Quantity of various cells was counted on specified smears views after staining. We observed estrous cycles of Rev-erb-α^+/+ and Rev-erb-α^-/- female mice using line plots and periodic spectrograms. The results showed that the Rev-erb-α^-/- female mice were infertility, and the number of embryos of Rev-erb-α^-/- females was less than that of Rev-erb-α^+/+ females. However, the follicle development of Rev-erb-α^-/- female mice was normal. The estrous cycle of Rev-erb-α^-/- female mice was 3.22 days longer than that of Rev-erb-α^+/+ female mice. The results suggest that loss of Rev-erb-α prolongs estrous cycle, which is probably one of the reasons for female mice infertility, and circadian rhythm is important for mammalian estrous cycle.

Key words: Rev-erb-α; estrous cycle; vaginal smear; circadian rhythm

收稿日期：2013-02-01　　录用日期：2013-04-12

通讯作者：高基民　　E-mail: jimingao64@163.com

引用本文：

张芹, 董珍, 吴曦, 颜洁, 高基民. 生物钟基因Rev-erb-α缺失导致雌鼠生理周期延长[J]. 生理学报 2013; 65 (4): 395-401.

ZHANG Qin, DONG Zhen, WU Xi, YAN Jie, GAO Ji-Min. [Circadian clock gene Rev-erb-α deficiency prolongs female estrous cyclicity.] [Article in Chinese]. Acta Physiol Sin 2013; 65 (4): 395-401 (in Chinese with English abstract).