Expression of Early Pregnancy Factors (EPF) in the Main Reproductive Organs of Female Yak (Bos grunnien)
ZHAO Sheng-Xian1, FAN Jiang-Feng1,*, YU Si-Jiu2, CUI Yan2, HAN Xiao-Hong2, MA Yue1, HE Hong-Hong2, MA Jin-Biao1, CAO Yan-Tao1, Bao Mei1
1 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China; 2 Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep ＆ Goat, Lanzhou 730070, China
Abstract：Early pregnancy factor (EPF), a kind of pregnancy related protein, plays an important role in the development of embryo as an autocrine and paracrine immune tolerance factor and growth factor of trophoblast layer. In this study, ovary, uterus, fallopian tube and placenta samples were collected respectively from female yak (Bos grunniens) during follicular phase, luteal phase and early pregnant period to investigate the expression of EPF and it role in reproduction. qRT-PCR, Western blott and immunohistochemistry (IHC) techniques were used to detect the expression of EFP mRNA and protein. The result indicated that EPF protein was most distributed in uterine glandular epithelial cells, endometrial epithelial cells, oviduct epithelial cells, membranous luteal cells, placental trophoblastic giant cells and crypt epithelial cells. qRT-PCR results showed that expression level of EPF mRNA in the ovary was significantly lower during follicular phase than that during early pregnant period and luteal phase. In oviduct and uterus the expression level of EPF mRNA was significantly higher during the follicular phase than that during the luteal phase and early pregnant period. The expression of EPF mRNA in ovarian and maternal placenta during early pregnant period was significantly higher than other reproductive organs. Western blot results showed that the expression of EPF protein in ovarian and oviduct during the early pregnant period was significantly higher than that during the luteal phase and follicular phase. In uterus of the follicular phase, the expression of EPF protein was significantly higher than that during the early pregnant period and luteal phase. It was noticeable that the expression of EPF in ovary and maternal placenta in early pregnant period was significantly higher than other reproductive organs, especially in the ovary. The results showed that EPF was spatiotemporal differently expressed in the main reproductive organs of female yak. During early pregnancy, EPF was mainly derived from ovary and maternal placenta. This study provides some useful materials to investigate the whole role of EPF in reproduction of yak.
赵生贤, 樊江峰, 余四九, 崔燕, 韩小红, 马悦, 何翃闳, 马进彪, 曹艳桃, 包梅. 早孕因子(EPF)在雌性牦牛主要生殖器官中的表达[J]. 农业生物技术学报, 2020, 28(5): 876-883.
ZHAO Sheng-Xian, FAN Jiang-Feng, YU Si-Jiu, CUI Yan, HAN Xiao-Hong, MA Yue, HE Hong-Hong, MA Jin-Biao, CAO Yan-Tao, Bao Mei. Expression of Early Pregnancy Factors (EPF) in the Main Reproductive Organs of Female Yak (Bos grunnien). 农业生物技术学报, 2020, 28(5): 876-883.
[1] 韩小红, 樊江峰, 于艺腾, 等. 2018.妊娠期和分娩前后牦牛胎盘组织形态学变化及细胞凋亡的研究[J]. 农业生物技术学报, 26(10): 1714-1722. (Han X H, Fan J F,Yu Y T.2018. Study on the morphological changes and apoptosis of yak (Bos grunniens) placentomes from pregnancy, pre and post-parturation[J]. Journal of Agricultural Biotechnology, 26(10): 1714-1722. [2] 何翃闳, 潘阳阳, 张慧珠, 等. 2018. FSH对牦牛卵母细胞EGF、EGFR表达及其细胞凋亡的影响[J]. 畜牧兽医学报, 49(09): 1899-1907. (He H H, Pan Y Y, Zhang H Z.2018. The Effects of follicle-stimulating hormone (FSH) on the expression of EGF and EGFR in yak oocytes and apoptosis[J]. Acta Veterinaria et Zootechnica Sinica, 49(09): 1899-1907. [3] 林燕棉. 2015.恶性生殖系统肿瘤组织中EPF的表达及EPF-McAb对生殖肿瘤细胞增殖的影响[D]. 硕士学位论文. 广州医科大学, 导师: 王家骥. pp. 30-32. (Lin Y M.2015. Expression of EPF in malignant tumors of the reproductive system and effect of EPA-McAb on reproductive tumor cells proliferation[D]. Thesis for M.S., Guangzhou Medical University, Supervisor:Wang J J,pp. 30-32.) [4] 王靖雷, 崔燕, 余四九, 等. 2019. 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, 27(03): 431-440. (Wang J L, Cui Y, Yu S J, et al.2019. Artemin expression and localization in main reproductive organs and parthenogenetic embryos of yak (Bos grunniens)[J]. Journal of Agricultural Biotechnology, 27(03): 431-440. [5] 张诗萌, 佟翠, 李鹏, 等. 2019. 早孕因子及其临床应用研究进展[J]. 黑龙江畜牧兽医, (05): 41-43. (Zhang S M, Tong C, Li P, et al. 2019. Research progress on early pregnancy factor and its clinical application[J]. Heilongjiang Animal Husbandry And Veterinary Medicine, (05): 41-43.) [6] Aluvihare V R, Kallikourdis M, Betz A G.2004. Regulatory T cells mediate maternal tolerance to the fetus[J]. Nature Immunology, 5: 266-271. [7] Bemis L T, McCue P M, Hatzel J N, et al.2012. Evidence for production of early pregnancy factor (Hsp10), microRNAs and exosomes by day 8 equine embryos[J]. Journal of Equine Veterinary Science, 32: 398. [8] Cavanagh A C, Morton H.1994. Early pregnancy factor: Purification to homogenity from human platelets and identification as chaperonoin 10[J]. European Journal of Biochemistry; 222: 551-560. [9] Chen Q G, Zhu X R,Chen R J, et al.2016. Early pregnancy factor enhances the generation and function of CD4+CD25+ regulatory T cells[J]. The Tohoku Journal of Experimental Medicine, 240(3): 215-220. [10] Corrao S,Campanella C, Anzalone R, et al.2009. Human Hsp10 and early pregnancy factor (EPF) and their relationship and involvement in cancer and immunity: Current knowledge and perspectives[J]. Life Sciences, 86(5-6): 145-152. [11] Dang-Nguyen T Q, Kaneda M, Somfai T, et al.2011. Development of single blastomeres derived from two-cell embryos produced in vitro in pigs[J]. Theriogenology, 76(1): 88-96. [12] Forde N, Bazer F W, Spencer Spencer T E.2015. Lonergan Pat. 'Conceptualizing' the endometrium: Identification of conceptus-derived proteins during early pregnancy in cattle[J]. Biology of Reproduction, 92(6): 156. [13] Gad A, Besenfelder U, Rings F, et al.2011. Effect of reproductive tract environment following controlled ovarian hyperstimulation treatment on embryo development and global transcriptome profile of blastocysts: Implications for animal breeding and human assisted reproduction[J]. Human Reproduction, 26(7): 1693-1707. [14] Grosso M C, Bellingeri R V, Motta C E, et al.2015. Immunohistochemical distribution of early pregnancy factor in ovary, oviduct and placenta of pregnant gilts[J]. Biotechnic & Histochemistry, 90(1): 14-24. [15] Grosso M C, Bellingeri R V, Schade R, et al.2012.Neutralization of early pregnancy factor by passive immunization alters normal embryonic development and cytokine balance[J]. Inmunologia, 31: 106-114. [16] Hatzel J N, Bouma G J, Cleys E R, et al.2015. Identification of heat shock protein 10 within the equine embryo, endometrium, and maternal peripheral blood mononuclear cells[J]. Theriogenology, 83(5): 832-839. [17] Hatzel J N, McCue P M, Ehrhart E J, et al.2012. Immunohistochemical localization of early pregnancy factor (Hsp10) in equine embryos[J]. Journal of Equine Veterinary Science, 32(7): 399. [18] Ito K, Takahashi M, Kawahata K, et al.1998. Supplementation effect of early pregnancy factor-positive serum into bovine in vitro fertilization culture[J]. American Journal of Reproductive Immunology. 39: 356-361. [19] Kawamura K, Fukuda J, Itoh H, et al.2000. Chaperonin 10 in the rat oocytes and early embryos: Its expression and activity for early pregnancy factor[J]. American Journal of Reproductive Immunology(Print), 44(4): 242-248. [20] Klein C.2016. Maternal recognition of pregnancy in the context of equine embryo transfer[J]. Journal of Equine Veterinary Science, 41: 22-28. [21] Mitchell L A, Nixon B, Aitken R J.2007. Analysis of chaperone proteins associated with human spermatozoa during capacitation[J]. Molecular Human Reproduction, 13(9): 605-613. [22] Morton H.1998. Early pregnancy factor: An extracellular chaperonin 10 homologue[J]. Immunology and Cell Biology, 76: 483-496. [23] Morton H, Hegh V, Clunie G.1974. Immunosuppression detected in pregnant mice by rosette inhibition test[J]. Nature, 249: 459-60. [24] Morton H, Rolfe B, Cavanagh A C.1992. Early pregnancy factor[J]. Seminars in Reproductive Endocrinology, 10(2): 72-82. [25] Nahhas F, Barnea E.1990. Human embryonic origin early pregnancy factor before and after implantation[J]. American Journal of Reproductive Immunology, 22(3-4): 105-108. [26] Ohnuma K, Yokoo M, Ito K, et al.2000. Study of early pregnancy factor (EPF) in equine (Equus caballus)[J]. American Journal of Reproductive Immunology (Print), 43(3):174-179. [27] Ott T L.2019. Symposium review: Immunological detection of the bovine conceptus during early pregnancy[J]. Journal of Dairy Science, 102(4): 3766-3777. [28] Sasaki Y, Sakai M.2004. Decidual and peripheral blood CD4+CD25+ regulatory T cells in early pregnancy subjects and spontaneous abortion cases[J]. Molecular Human Reproduction, 10: 347-353. [29] Vasudevan S, Kamat M M, Walusimbi S S, et al.2017. Effects of early pregnancy on uterine lymphocytes and endometrial expression of immune-regulatory molecules in dairy heifers[J]. Biology of Reproduction, 97(1): 104-118. [30] Walsh A,Whelan D, Bielanowicz A, et al.2008. Identification of the molecular chaperone, heat shock protein 1 (chaperonin 10), in the reproductive tract and in capacitating spermatozoa in the male mouse[J]. Biology of Reproduction, 78(6): 983-993. [31] Wang W J, Liu F J, Zhang X, et al.2017. Periodic elevation of regulatory T cells on the day of embryo transfer is associated with better in vitro fertilization outcome[J]. Journal of Reproductive Immunology, 119: 49-53.
