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2025年8月8日 星期五
农业生物技术学报  2019, Vol. 27 Issue (3): 431-440    DOI: 10.3969/j.issn.1674-7968.2019.03.006
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
Artemin在牦牛主要生殖器官和孤雌激活胚胎中的表达及定位
王靖雷, 王萌, 潘阳阳, 李秦, 何翃闳, 胡学权, 韩小红, 樊江峰, 崔燕, 余四九*
甘肃农业大学 动物医学院/甘肃省牛羊胚胎工程技术研究中心,兰州 730070
Artemin Expression and Localization in Main Reproductive Organs and Parthenogenetic Embryos of Yak (Bos grunniens)
WANG Jing-Lei, WANG Meng, PAN Yang-Yang, LI Qin, HE Hong-Hong, HU Xue-Quan, HAN Xiao-Hong, FAN Jiang-Feng, CUI Yan, YU Si-Jiu*
College of Veterinary Medicin/Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep & Goat, Gansu Agricultural University, Lanzhou 730070, China
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摘要 Artemin (ARTN)是胶质细胞源神经营养因子(glial cell line-derived neurotrophic factor, GDNF)配体家族成员之一,在多种哺乳动物生殖道和早期胚胎发育过程中表达并参与早期胚胎的发育调节,但尚未明确其是否存在于牦牛(Bos grunniens)主要生殖器官和胚胎中。为探究正常生理条件下Artemin在牦牛发情周期及早期胚胎发育过程中的调控作用及生物学机制,本研究分别采集不同繁殖周期(卵泡期, 黄体期及妊娠期)牦牛的主要生殖器官(输卵管, 卵巢, 子宫),以及孤雌激活胚胎,利用qRT-PCR检测Artemin基因在组织和胚胎发育过程中的表达,同时利用蛋白质免疫印迹(Western blot, WB)、免疫组织化学方法以及免疫荧光染色方法,分析Artemin在各生殖器官和胚胎中的表达水平与定位。结果显示,Artemin在牦牛主要生殖器官和孤雌激活胚胎中均有表达,其中黄体期与妊娠期的输卵管和卵巢中Artemin表达均显著高于卵泡期;卵泡期和妊娠期子宫中Artemin的表达显著高于黄体期;孤雌激活胚胎中Artemin的表达在桑椹胚期最高,4~8-细胞与囊胚期次之,2-细胞期最低。免疫组织化学结果显示,Artemin在输卵管黏膜上皮、卵泡膜、卵巢生殖上皮、黄体细胞、子宫内膜和子宫腺体均有表达。免疫荧光染色结果显示,Artemin在牦牛囊胚滋养层有表达。本研究结果表明,Artemin参与牦牛发情周期和早期胚胎发育的调控作用。该研究结果为进一步探究Artemin的作用机制提供了基础资料,有助于牦牛胚胎体外培养技术的改进和牦牛繁殖能力的提高。
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作者相关文章
王靖雷
王萌
潘阳阳
李秦
何翃闳
胡学权
韩小红
樊江峰
崔燕
余四九
关键词 Artemin (ARTN)牦牛繁殖周期滋养层    
Abstract:Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) ligand family, expressed in a variety of mammalian reproductive tract and early embryo development, and involved in early embryonic developmental regulation. But the presence of Artemin in the main reproductive organs and embryos of the yak (Bos grunniens) is unknown. In order to explore the biological effects of Artemin in female yak reproductive regulation and early embryo development under normal physiological conditions, this study collected the main reproductive organs (ovary, uterus, fallopian tubes) of female yak at different stages (follicular phase, luteal phase, and pregnancy) and produced parthenogenetically activated embryos. qRT-PCR was used to detect the expression of Artemin gene during tissue and embryonic development. Western blot(WB), immunohistochemistry and immunofluorescence staining were used to analyse the Artemin expression level and localization in various reproductive organs and embryos. The results showed that Artemin was expressed in the main genital organs and parthenogenetic embryos of yak. The expression of Artemin in the luteal phase and the oviduct during pregnancy was higher than that in the follicular phase. The expression of Artemin in the uterus during follicular phase and pregnancy was higher than that in the luteal phase. The levels of Artemin mRNA in embryos were higher than that in main genital, which were highest in morula, followed by 4~8-cell embryos and blastocyst , and were lowest in 2-cell embryos. Immunohistochemistry results showed that Artemin was expressed in the oviductal mucosa epithelium, follicular membrane, ovarian reproductive epithelium, luteal cells, endometrium and uterine glands. Immunofluorescence staining showed that Artemin was expressed in the trophoblast of the yak blastocyst. These results showed that Artemin was involved in the regulation of yak estrous cycle and early embryo development. This study provides basic data for further exploration of the mechanism of action of Artemin, which contributes to the improvement of in vitro culture techniques of yak embryos and the improvement of yak breeding ability.
Key wordsArtemin (ARTN)    Yak    Reproductive cycle    Trophoblast
收稿日期: 2018-09-05     
ZTFLH:  S823.8+5.2  
基金资助:国家重点研发项目(No. 2017YFD0502200)、国家自然科学基金(No. 31472244)、国家自然基金地区基金(No. 31660732)、国家自然科学基金青年科学基金(No. 31702311)、甘肃省青年科技基金计划(No. 17JR5RA141)和甘肃省重点研发项目(No. 17YF1NA058)
通讯作者: *sjyu@163.com   
引用本文:   
王靖雷, 王萌, 潘阳阳, 李秦, 何翃闳, 胡学权, 韩小红, 樊江峰, 崔燕, 余四九. Artemin在牦牛主要生殖器官和孤雌激活胚胎中的表达及定位[J]. 农业生物技术学报, 2019, 27(3): 431-440.
WANG Jing-Lei, WANG Meng, PAN Yang-Yang, LI Qin, HE Hong-Hong, HU Xue-Quan, HAN Xiao-Hong, FAN Jiang-Feng, CUI Yan, YU Si-Jiu. Artemin Expression and Localization in Main Reproductive Organs and Parthenogenetic Embryos of Yak (Bos grunniens). 农业生物技术学报, 2019, 27(3): 431-440.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.03.006     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I3/431
 
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