牦牛CAV1基因克隆及其在雌性生殖系统主要器官中的表达定位  

Cloning of Yak(Bos grunniens) CAV1 Gene and Its Expression in Major Organs of Female Reproductive System

作　　者：张瑞 王靖雷 潘阳阳 王静瑜 胡学权 马睿 崔燕 余四九 ZHANG Rui;WANG Jing-Lei;PAN Yang-Yang;WANG Jing-Yu;HU Xue-Quan;MA Rui;CUI Yan;YU Si-Jiu(College of Veterinary Medicine/Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep&Goat,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]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

出　　处：《农业生物技术学报》2020年第4期681-692,共12页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31702311);甘肃省重点研发项目(17YF1NA058);甘肃省教育厅创新能力提升项目(2019B-081);甘肃省教育厅产业支撑引导项目(2019C-03);甘肃农业大学青年伏羲基金项目(Gaufx‐02Y10)。

摘　　要：窖蛋白-1(Caveolin-1, CAV1)是一种与内吞作用、细胞外基质组织、胆固醇分布、细胞迁移和信号传导相关的膜蛋白。为探索正常生理条件下CAV1在雌性牦牛(Bos grunniens)生殖系统中的表达及其生理功能,本研究以牦牛不同繁殖时期(卵泡期,黄体期和妊娠期)的子宫、卵巢和输卵管为材料,利用基因克隆、qRT-PCR、蛋白质免疫印迹(Western blot)、免疫组织化学(immunohistochemistry, IHC)等方法,对CAV1基因和蛋白的表达进行分析。结果显示,CAV1基因(GenBank No. MN127963)高度保守,牦牛CAVI基因序列与普通牛(Bos taurus)相比有差异,但其编码的氨基酸不变,牦牛CAVI基因与普通牛和绵羊(Ovis aries)亲缘性最近,与家猫(Felis catus)和家犬(Canis lupus familiaris)亲缘性最远,其编码的蛋白为稳定的疏水性膜蛋白;CAV1在雌性牦牛生殖系统主要器官中均有表达,其中卵泡期输卵管、卵巢、子宫中CAV1的表达水平显著高于黄体期、妊娠期(P<0.05),妊娠期输卵管中CAV1的表达水平显著高于黄体期(P<0.05),黄体期卵巢和子宫中CAV1的表达水平显著高于妊娠期(P<0.05);免疫组织化学结果显示,CAV1在输卵管黏膜上皮、浆液腺,卵巢卵泡膜、卵泡颗粒层、生殖上皮、黄体细胞,子宫内膜和子宫腺体均有表达。研究结果为深入探索CAV1参与哺乳动物生殖生理调控提供了理论基础,有助于高原哺乳动物繁殖规律的研究。Caveolin-1(Caveolin-1, CAV1) is a membrane protein associated with endocytosis, extracellular matrix organization, cholesterol distribution, cell migration and signaling.The aim of the present study was to explore the expression and biological function of CAV1 in the reproductive system of female yak(Bos grunniens) under normal physiological conditions. In this study, the uterus, ovaries and fallopian tubes of female yaks at different reproductive stages(follicula, luteal and gestation) were used as materials. The mRNA and protein expression of CAV1 were analyzed by gene cloning, qRT-PCR, Western blot and immunohistochemistry(IHC). The results showed that the CAV1 gene(GenBank No. MN127963) was highly conserved. The CAVI gene sequence of yak was different from ordinary cattle(Bos taurus), but the encoded amino acid was constant. The CAVI gene of yak had the closest relatives to Bos taurus and Ovis aries, and the farthest relatives to Felis catus and Canis lupus familiaris, and the encoded protein was a stable hydrophobic membrane protein. CAV1 was expressed in the main organs of the female yak reproductive system, while the expression level of CAV1 protein in the oviduct, ovary and uterus in the follicular phase was significantly higher than that in the luteal phase and gestational period(P<0.05). The expression level of CAV1 protein in the fallopian tube of the gestational period was significantly higher than that in the luteal phase(P<0.05). The expression level of CAV1 protein in the ovary and uterus of the luteal phase was significantly higher than that in the gestation phase(P<0.05). Immunohistochemistry results showed that CAV1 was expressed in the oviduct mucosa epithelium, serous gland, ovarian follicle membrane, follicular granule layer, luteal cells, reproductive epithelium, endometrium and uterine gland. This study provides a theoretical basis for further exploration of CAV1 involved in mammalian reproductive physiology regulation, which is helpful to the study of the breeding rules of mammals in the p

关 键 词：牦牛 窖蛋白-1(CAV1) 胞吞 

分 类 号：S823.85.2[农业科学—畜牧学]