甘加型藏羊发情周期和乏情期血浆NKB动态变化及HPO轴NK3R的表达  

Dynamic Changes of NKB in Plasma and Expression of NK3R in HPO Axis during Estrous Cycle and Anestrus of Ganjia Tibetan Sheep

作　　者：赵国勇 高何璇 刘晓洁 何玉琴[1] 陶乐凯 蔡永强 张光敏 ZHAO Guoyong;GAO Hexuan;LIU Xiaojie;HE Yuqin;TAO Lekai;CAI Yongqiang;ZHANG Guangmin(College of Life Science and Technology,Gansu Agricultural University,Lanzhou,Gansu 730070;Network Finance Department,Bank of Lanzhou,Lanzhou,Gansu 730070)

机构地区：[1]甘肃农业大学生命科学技术学院,甘肃兰州730070 [2]兰州银行网络金融部,甘肃兰州730070

出　　处：《核农学报》2024年第9期1699-1706,共8页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金资助项目(31960689)。

摘　　要：为探讨神经激肽B(NKB)和神经激肽B受体(NK3R)在甘加型藏羊(Ovis aries)繁殖调控中的作用,本研究采用酶联免疫吸附法(ELISA)、实时荧光定量PCR(qRT-PCR)、蛋白免疫印迹(western blot)和免疫组织化学(IHC)方法,对甘加型藏羊发情周期和乏情期血浆NKB含量、下丘脑-垂体-卵巢(HPO)轴组织NK3R mRNA及蛋白的表达和分布进行检测。结果表明,甘加型藏羊发情周期和乏情期血浆中NKB含量呈动态变化,NKB平均含量在发情期最低、间情期最高,二者差异显著(P<0.05),发情周期各时期的NKB含量均高于乏情期。发情周期和乏情期HPO轴组织均有NK3R mRNA和蛋白表达,且各时期表达存在差异,下丘脑中发情前期NK3R mRNA表达量最高,显著高于发情期、发情后期、间情期和乏情期(P<0.05),NK3R蛋白在发情后期表达量最高;垂体中间情期NK3R mRNA表达量最高,与其他时期差异显著(P<0.05),NK3R蛋白在发情期表达量最高;卵巢组织中NK3R mRNA和蛋白表达趋势一致,均在发情前期表达量高;NK3R蛋白主要分布于下丘脑神经内分泌小细胞和神经胶质细胞,垂体嗜酸性细胞、嫌色细胞、嗜碱性细胞,卵巢颗粒层细胞、卵泡膜、黄体细胞中。以上结果表明,NKB与HPO轴特异性受体NK3R结合,参与调节甘加型藏羊生殖内分泌和周期性发情等生殖活动。本研究结果可为深入探究藏羊繁殖调控机理提供科学依据。To investigate the role of Neurokinin B(NKB)and Neurokinin B receptor(NK3R)in the regulation of reproduction in Ganjia Tibetan sheep(Ovis aries).In this study,the methods of ELISA,quantitative real-time PCR(qRT-PCR),western blot and immunohistochemistry(IHC)were used to detect the NKB content in plasma,and the expression and distribution of NK3R mRNA and protein in hypothalamuspituitary-ovary(HPO)axis tissues during the estrous cycle and anestrus of Ganjia Tibetan sheep.The results showed that the NKB content of plasma changed dynamically during estrous cycle and anestrus of Ganjia Tibetan sheep,with the lowest average NKB content in the estrus and the highest average NKB content in the diestrus,showing a significant difference(P<0.05).The NKB content in plasma during each estrous cycle period was higher than in anestrus.The expression of NK3R mRNA and protein in HPO axis tissues during the estrous cycle and anestrus of Ganjia Tibetan sheep varied among different periods.In hypothalamus,the expression of NK3R as the highest in the proestrus,which was significantly higher than that in estrus,metestrus,diestrus and anestrus(P<0.05),while the highest NK3R protein level was found in the metestrus.In pituitary,the expression of NK3R was the highest in the diestrus,significantly differed from that of the other periods(P<0.05),and the highest expression of NK3R protein was found in the estrus.The expression patterns of NK3R mRNA and protein in ovary were similar,both peaking in the proestrus.NK3R protein was predominantly distributed in neuroendocrine small cells and glial cells of the hypothalamus,acidophilic cells,chromophobe cells,and basophilic cells of pituitary,as well as the stratum granulosum,theca folliculi,and luteal cells of the ovary.These findings indicate that NKB binds to the HPO axis-specific receptor NK3R,playing a role in regulating reproductive activities such as reproductive endocrinology and cyclic estrus in Ganjia Tibetan sheep.The results provide a scientific foundation for further exploration of

关 键 词：甘加型藏羊 神经激肽B 神经激肽B受体 下丘脑-垂体-卵巢轴 发情周期 

分 类 号：S826[农业科学—畜牧学]