调控CTSK基因的候选miRNA在单羔、多羔贵州黑山羊卵巢和子宫体的表达分析  

Analysing the expression of the candidate miRNA regulating the CTSK gene in the ovary and uterine tissues of single-lamb and multi-lamb Guizhou black goats

作　　者：陆情梅 杨永鲜 冉铭 刘彬[1,2] 周明帅[1,2] 温晓艳 赵佳福 LU Qingmei;YANG Yongxian;RAN Ming;LIU Bin;ZHOU Mingshuai;WEN Xiaoyan;ZHAO Jiafu(College of Animal Science,Guizhou University,Guiyang 550025,China;Key Laboratory of Genetic Breeding and Reproduction of Plateau Mountain Animals of Ministry of Education,Guizhou University,Guiyang 550025,China;Guizhou Giles Circular Agriculture Co.,Ltd.,Qiannan Buyei and Miao Autonomous Prefecture 558110,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州大学高原山地动物遗传育种与繁殖教育部重点实验室,贵州贵阳550025 [3]贵州省吉尔斯循环农业有限公司,贵州黔南布依族苗族自治州558110

出　　处：《畜牧与兽医》2023年第5期7-12,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家级大学生创新创业训练计划项目(202110657012);贵州省科技计划项目(黔科合支撑[2022]一般089);黔南州农业攻关项目(黔南科合[2021]27号)。

摘　　要：以贵州黑山羊为研究对象,通过生物在线软件预测调控CTSK基因表达的候选miRNAs,采用荧光定量PCR法检测CTSK及其候选miRNAs在贵州黑山羊单、多羔性腺轴下丘脑、垂体、卵巢、输卵管、子宫体及子宫角6种组织中的表达量。结果:CTSK在单、多羔贵州黑山羊的6个组织中均有表达,在单羔组中表达量依次为子宫角>输卵管>卵巢>子宫体>垂体>下丘脑,其中在子宫角中的表达量显著高于其他组织(P<0.05),在多羔组中,其表达量依次为子宫体>输卵管>子宫角>卵巢>垂体>下丘脑,其中子宫体的表达量显著高于其他组织(P<0.05);组间比较可知,CTSK基因在多羔组子宫体中的表达量极显著高于单羔组(P<0.01),卵巢中的表达量极显著低于单羔组(P<0.01)。在线软件预测结果表明:miR-122、miR-205、miR-7、miR-30-5p和miR-124为调控CTSK表达评分最高的5条miRNAs。qPCR结果表明:5条miRNAs在子宫体和卵巢中均有表达,在子宫体中,单羔组和多羔组的表达量均依次为miR-122>miR-30-5p>miR-205>miR-7>miR-124,其中miR-122的表达量显著高于其他miRNAs(P<0.05);组间比较显示,单羔组中miR-122、miR-30-5p、miR-205和miR-124表达量极显著高于多羔组(P<0.01),而miR-7表达量显著高于多羔组(P<0.05)。在卵巢组织中,5条miRNAs在单羔组中的表达量依次为miR-30-5p>miR-122>miR-205>miR-7>miR-124,其中miR-30-5p的表达量显著高于其他miRNAs(P<0.05),在多羔组中,其表达量依次为miR-122>miR-30-5p>miR-7>miR-205>miR-124,其中miR-122的表达量显著高于其他miRNAs(P<0.05),而miR-124表达量最低;组间比较可知,miR-7在多羔组中极显著高于单羔组(P<0.01),而miR-30-5p在多羔组中极显著低于单羔组(P<0.01)。本研究解析了CTSK基因在贵州黑山羊性腺轴上的表达情况,筛选到5条调控CTSK表达的候选miRNAs,弄清了5条miRNAs在贵州黑山羊子宫体和卵巢组织上的表达谱,为进一步阐明miRNA调控CTSK基因影响山羊繁殖率的分子�Guizhou black goats were used as the research subjects to study the expression of the CTSK gene and its candidate miRNAs in the gonadal axis,and the candidate miRNAs for regulating the expression of CTSK gene were predicted by biological online software,and the expression levels of the CTSK gene and its candidate miRNAs in hypothalamus,pituitary,ovary,fallopian tube,uterine body,and uterine horn of Guizhou black goats were detected by fluorescence.CTSK was found in all the six tissues of the Guizhou black goats with single-and multiple-lambs.The expression level in the different tissues of in the single lamb group was in the order of the uterine horn>fallopian tube>ovary>uterine body>pituitary gland>hypothalamus,and the expression level in the uterine horn was significantly higher than in the other tissues(P<0.05).In the multiple-lamb group,the expression level was in the order of the uterine body>fallopian tube>uterine horn>ovary>pituitary gland>hypothalamus;and the expression level in the uterine body was significantly higher than in the other tissues(P<0.05).By group comparison,the expression of the CTSK gene in the uterus of the multiple-lamb group was significantly higher than that of the single-lamb group(P<0.01);however,the expression in the ovary was significantly lower(P<0.01).According to the results of online software prediction,the five miRNAs with the highest scores in regulating CTSK expression are miR-122,miR-205,miR-7,miR-30-5p,and miR-124.The five miRNAs were found to express in the uterus and ovary,according to the qPCR results.In the uterine body,the expression levels of the single-lamb group were in the order of miR-122>miR-30-5p>miR-205>miR-7>miR-124,with the expression of miR-122 being significantly higher than the other miRNAs(P<0.01).The expression level in the multiple-lamb group was in the order of miR-122>miR-30-5p>miR-205>miR-7>miR-124,and the expression of miR-122 was also significantly higher than the other miRNAs(P<0.05).The expression levels of miR-122,miR-30-5p,miR-205,and miR-124

关 键 词：贵州黑山羊 CTSK基因 MICRORNA 特异性表达 

分 类 号：S813[农业科学—畜牧学]