IL-1β及其受体在雌牦牛主要生殖器官和孤雌激活胚胎中的表达定位  

Expression and Localization of Interleukin 1 Beta and Interleukin 1 Receptor(Type Ⅰ)in Main Reproductive Organs and Parthenogenetic Embryos of Female Yak(Bosgrunniens)

作　　者：王靖雷 王萌 潘阳阳 李秦 何翃闳 黄玉风 赵凌 樊江峰 崔燕 余四九 WANG Jinglei;WANG Meng;PAN Yangyang;LI Qin;HE Honghong;HUANG Yufeng;ZHAO Ling;FAN Jiangfeng;CUI Yan;YU Sijiu(Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep&Goat,College of Veterinary Medicine,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：《西北农业学报》2018年第10期1395-1404,共10页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(31472244;31702311);甘肃省青年科技基金计划(17JR5RA141);甘肃省科技厅项目(1506RJDA002)~~

摘　　要：为探索正常生理条件下IL-1β和IL1R1参与雌性牦牛生殖调控及早期胚胎发育的生物学作用,试验采集雌性牦牛主要生殖器官(卵泡期:输卵管、卵巢、子宫;黄体期:输卵管、卵巢、子宫;妊娠期:输卵管、卵巢、子宫),并生产孤雌激活胚胎。实时荧光定量PCR(Quantitative real-time PCR,qRT-PCR)检测IL-1β和IL1R1 mRNA在组织和胚胎发育过程中的表达;并采用蛋白质免疫印迹(Western-blot,WB)和免疫组织化学方法从蛋白水平分析IL-1β和IL1R1在各生殖器官的表达水平与定位。结果显示IL-1β和IL1R1在牦牛主要生殖器官和孤雌激活胚胎中均有表达。其中卵泡期输卵管中IL-1β和IL1R1表达均高于黄体期和妊娠期,卵泡期卵巢和妊娠期卵巢中IL-1β和IL1R1的表达高于黄体期,妊娠期子宫中IL-1β和IL1R1的表达高于卵泡期和黄体期,孤雌激活胚胎中IL-1β和IL1R1的表达桑椹胚最高,8细胞次之,2细胞和囊胚期最低。免疫组织化学结果显示IL-1β和IL1R1在输卵管黏膜上皮、卵泡颗粒层、黄体细胞、子宫上皮和子宫腺体均有表达。研究结果表明IL-1β和IL1R1参与牦牛生殖周期和胚胎发育的调控作用,为进一步探究IL-1β和IL1R1的作用机制提供了理论基础,有助于牦牛胚胎体外培养技术的改进。The aim of this study was to explore the biological function of IL-1βand IL1R1 in main reproductive organs of female yak during early embryonic development.The main reproductive organs(follicular phase:fallopian tubes,ovaries,uterus;luteal phase:fallopian tubes,ovaries,uterus;pregnancy:fallopian tubes,ovaries,uterus)of female yak were collected under the normal physiological conditions,and parthenogenetic activationembryos were also produced and cultured in vitro.The levels of IL-1β,IL1R1 mRNA in different tissues and embryos were detected by Quantitative Real-time PCR(qRT-PCR).Moreover,the levels and location of IL-1βand IL1R1 proteins were detected by methods of Western-blot and Immunohistochemistry.Our results showed that mRNA of IL-1βand IL1R1 could be detected in all reproductive organs and embryos at different stages.However,the levels of IL-1βand IL1R1 in the fallopian tube at follicular phase was higher than those at luteal phase and gestation phase.Secondly,the levels of IL-1βand IL1R1 in ovarian follicle and ovary during pregnancy was higher than those in the luteal phase.IL-1βand IL1R1 levels in uterus during pregnancy were higher than those in inuterus at luteal phase and follicular phase.The levels of IL-1βand IL1R1 mRNA in embryos were higher than those in main genital,which were highest in morula,followed by 8-cell embryos and 2-cell embryos,and was lowest in blastocyst.Immunohistochemical results showed that IL-1βand IL1R1 could be observed on the mucosal epithelium,follicle granular layer,luteal cells,uterine epithelium and uterine glands.The above results indicate that IL-1βand IL1R1 play various biological functions in the reproductive process of yak,which might involve in oocyte maturation,ovulation,early embryonic and fetal development of gestational calves.The results provide theoretical basis for further exploration of the mechanism of IL-1βand IL1R1,and help to improve the in vitro culture technology of yak embryos.

关 键 词：牦牛 IL-1Β IL1R1 孤雌胚胎 

分 类 号：S823.85.2[农业科学—畜牧学]