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
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.
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.
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