Abstract:The regulation of angiogenic factors in the ovary of sheep (Ovis aries) is closely related to vascular proliferation, follicular development and corpus luteum formation, and has an important regulatory effect on the reproductive performance of sheep. In this study, qRT-PCR was used to detect the expression of VEGF gene in different ovarian ovary, and the expression and distribution of VEGF protein in ovarian tissue was analyzed by Immunohistochemistry and Western blot combined with IPP(image pro plus) technology. The results showed that VEGF was expressed in sheep ovary, and the expression of high fertility sheep was significantly higher than that of low fertility sheep (P<0.05). Studies showed that VEGF-positive signals were mainly localized in granulosa cells, luteal cells and vascular endothelial cells in different follicles of sheep, and were also weakly expressed in ovarian stroma and atresia follicular granulosa cells, the integrated optical density analysis of VEGF found that The expression of VEGF in the ovarian tissues of Small-tailed Han sheep, Hu sheep and Tibetan sheep was consistent. During the development of different follicles, the integrated optical density values of the follicles gradually increased and synchronized with follicular development. The positive signal strength was consistent with the follicle size, and the expression in the early stage of luteal formation was significantly higher than that in the degenerative stage. Researches found that high expression of VEGF could induce more angiogenesis around the follicles and corpus luteum in sheep, which improved the permeability of microvessels and capillaries and played an active role in increasing the number and quality of dominant follicles. This study provides basic information for revealing some of the genetic mechanisms of differences in sheep fecundity.
郭亚军, 王欣荣. VEGF在不同繁殖力绵羊卵巢组织中的分布及表达[J]. 农业生物技术学报, 2020, 28(5): 866-875.
GUO Ya-Jun, WANG Xin-Rong. Distribution and Expression of VEGF in Ovarian Tissues of Sheep (Ovis aries) with Different Fertility. 农业生物技术学报, 2020, 28(5): 866-875.
[1] 曹彩霞. 2005. 缬沙坦对糖尿病肾病大鼠肾小球VEGF及其受体表达的影响[D]. 硕士学位论文, 青岛大学, 导师: 刘丽秋, pp. 10-20. (Cao C X.2005. Effects of valsartan on the expressions of vascular endothelial growth factor and its receptor in the kidney of diabetic nephropathy Rats[D]. Thesis for M. S., Qingdao University,Supervisor: Liu L Q, pp.10-20.) [2] 曹俊国, 陈敏, 李文, 等. 2018. 哺乳动物卵泡发育调控分子机制研究进展[J]. 特产研究, (4): 14-118. (Cao J G, Chen M, Li W, et al. 2018. Research progresses on the molecular mechanismsun derlying mammalian follicle development[J]. Special Wild Economic Animal and Plant Research, (4): 114-118.) [3] 成志军, 陶勇, 章孝荣. 2008. 胎猪卵巢发生过程中的组织学变化[J]. 细胞生物学杂志, (03): 352-356. (Cheng Z J, Tao Y, Zhang X R. 2008. Histological change during fetal pig ovarigenesis[J], Chinese Journal of Cell Biology,(03):352-356. ) [4] 黄颖, 周刚, 谢婷, 2008. 宣痹汤对Ⅱ型胶原诱导的关节炎大鼠关节组织VEGF的影响[J]. 安徽中医药大学学报,27(2): 27-29. (Hang Y, Zhou G, Xie T.2008. Effects of xuanbi decoction on expression of vascular endothelial growth factor in rats with collage-induced arthritis[J]. Journal of Anhui Traditional Chinese Medical College, 27(2): 27-29.) [5] 戢爽. 2007. VEGF基因在妊娠期绵羊主要生殖器官表达的研究[D]. 硕士学位论文, 吉林农业大学, 导师:姜怀志,pp.12-20. (Jian S.2007. The study on the expression of the vascular endothelial growth factor in reproductive organs of female ovine during pregnancy[D]. Thesis for M. S., Jilin Agriculture University, Supervisor: Jiang H Z, pp. 13-23.) [6] 时小艳. 2016, 血管生长调节因子与卵巢发育的关系[J]. 西北农业学报, 25(12): 1755-1761. (Shi X Y.2016. Relation of vascular endothelial growth actor and ovarian development[J]. Acta Agriculturae Boreali-occidentalis Sinica, 25(12): 1755-1761.) [7] 王典, 陈洋, 杨雨江, 等.2012. 绵羊发情周期中VEGF 在卵巢中的表达[J]. 中国兽医学报, 32(01): 151-156. (Wang D, Chen Y, Yang Y J, et al.2012. Expression of VEGF in ovary of ovine through the estrous cycle[J]. Chinese Journal of Veterinary Science, 32(01): 151-156.) [8] 王欣荣, 杨雅楠, 梁翕, 等. 2017. 绵羊卵巢动脉及其分支的解剖学特征[J]. 畜牧兽医学报, 48(08): 1452-1458. (Wang X Y, Yang Y N, Liang X, et al.2017. The anatomical characteristics of ovarian artery and its branches in sheep[J]. Chinese Journal of Animal and Veterinary Sciences, 48(08): 1452-1458.) [9] 杨雅楠, 刘成泽, 王彪, 等. 2018. 绵羊VEGF基因与Ang/Tie-2信号通路在卵巢中的表达分析[J]. 农业生物技术学报,2018, 26(07): 1219-1225. (Yang Y N, Liu C Z, Wang B, et al.2018. Expression analysis of VEGF gene and Ang/Tie-2 pathway in ovaries of sheep (Ovis aries)[J]. Journal of Agricultural Biotechnology, 26(07): 1219-1225.) [10] 雍艳红, 2003. 牦牛妊娠期卵泡和黄体的组织形态学研究[D]. 硕士学位论文, 甘肃农业大学, 导师: 崔燕, pp. 12-20. (Yong H Y.2003. Studies follicular the histological characteristics of corpus luteum in yaks during pregnancy[D]. Thesis for M. S., Gansu Agricultural University, Supervisor: Cui Y, pp. 13-23.) [11] 张慧慧, 宋炎成, 蔡道章. 2008. 基因多态性和股骨头坏死的相关研究进展[J]. 中国矫形外科杂志, (11): 833-836. (Zhang H H, Song Y C, Cai D Z. 2008. Advances in gene polymorphism and femoral head necrosis[J]. Orthopedic Journal of China, (11): 833-836) [12] 赵有璋. 2011. 羊生产学[M]. 中国农业出版社, 北京, pp. 125-127. (Zhao Y Z.2011. Sheep and Goats Production [M]. China Agricultural Press, Beijing, pp. 125-127.) [13] 郑惠丽, 倪江. 2004. 血管内皮生长因子(VEGF)与黄体形成和功能关系的研究概况[J]. 生殖与避孕, 24(4): 249-253. (Zheng H L, Ni J.2004. Overview of the relationship between vascular endothelial growth factor (VEGF) and luteal formation and function[J]. Reproduction and Contraception, 24(4): 249-253.) [14] Barboni B, Turriani M, Galeati G, et al.2000. Vascular endothelial growth factor production in growing pig antral follicles[J]. Biology of Reproduction, 63: 858-864. [15] Bruno J B, Celestino J J H, Lima-Verde I B, et al.2009. Expression of vascular endothelial growth factor (VEGF) receptor in goat ovaries and improvement of in vitro caprine preantral follicle survival and growth with VEGF[J]. Reproduction Fertility Development, 21(5): 679-687. [16] Chowdhury M W H, Scaramuzzi R J, Wheeler-Jones C P D, et al.2010. The expression of angiogenic growth factors and their receptors in ovarian follicles throughout the estrous cycle in the ewe[J]. Theriogenology, 73(7): 856-872. [17] Kaczmarek M M, Kiewisz J, Schams D, et al.2009. Expression of VEGF-receptor system in conceptus during peri-implantation period and endometrial and luteal expression of soluble VEGFR-1 in the pig[J]. Theriogenology, 71(8): 1298-1306. [18] Kawano Y, Zeineh H K, Fukuda J, et al.2003. Production of vascular endothelial growth factor and angiogenic factorin human follicular fluid[J]. Molecular and Cellular Endocrinology, 202: 19-23. [19] Kashida S,Sugino N, Takiguchi S, et al.2001. Regulation and role of vascular endothelial growth factor in the corpus luteum during mid-pregnancy in rats[J]. Biology of Reproduction, 64(1): 317-323. [20] Ntala V, Asimakopoulos B, Veletza S, et al.2015. Follicular fluid levels of vascular endothelial growth factor and its receptors and pregnancy outcome of women participating in intracytoplasmic sperm injection cycles[J]. Clinical & Experimental Obstetrics & Gynecology, 42(4): 437. [21] Rabbani M, Rogers P.2001. Role of vascular endothelial growth factor in endometrial vascular events before implantation in rats[J]. Reproduction, 122(1): 85-90. [22] Otani N, Minami S, Yamoto M, et al.1999. The vascular endothelial growth factor/fms-like tyrosine kinase systemin human ovary during the menstrual cycle and earlypregnancy[J]. The Journal of Clinical Endocrinology and Metabolism, 84(10): 3845-3851. [23] Shimizu T, Iijima K, Miyabayashi K, et al.2007. Effect of direct ovarian injection of vascular endothelial growth factor gene fragments on follicular development in immature female rats[J]. Reproduction, 134(5): 677-682. [24] Smith S K.2001. Angiogenesis and reproduction[J]. BJOG: An International Journal of Obstetrics & Gynaecology, 108(8): 777-783. [25] Stouffer R L, Martinez- Chequer J C, Molskness T A, et al.2001. Regulation and action of angiogenic factors in theprimate ovary[J]. Archives of Medical Research, 32(6): 567-575.