牦牛GnIH基因生物学特征分析及其在卵巢颗粒细胞中亚细胞定位的研究  

Biological characteristics analysis of yak gnih gene and study on its subcellularlocalization in ovarian granule cells

作　　者：胡宇磊 杨满珍 于海玲 朱艳锦 潘帮婷 付伟 熊燕[1] 李键[1] 熊显荣[1,2] HU Yulei;YANG Manzhen;YU Hailing;ZHU Yanjin;PAN Bangting;FU Wei;XIONG Yan;LI Jian;XIONG Xianrong(Ministry of Education Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Breeding Resource Reservation and Utilization,Southwest Minzu University,Chengdu 610041,China;State Ethnic Affairs Commission Key Laboratory of Animal Science,Southwest Minzu University,Chengdu 610041,China)

机构地区：[1]西南民族大学青藏高原动物遗传育种资源保护与利用国家教育部重点实验室,成都610041 [2]西南民族大学动物科学国家民委重点实验室,成都610041

出　　处：《黑龙江畜牧兽医》2023年第19期121-127,142,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：四川省科技厅重点研发计划项目(2021YFN0121);西南民族大学中央高校基本科研业务费项目(2021PTJS20)。

摘　　要：为探究促性腺激素抑制激素(gonadotropin-inhibitory hormone, GnIH)基因的序列特征及其在牦牛卵巢颗粒细胞中的表达与定位,试验从牦牛卵巢组织cDNA中克隆了GnIH基因,利用在线软件进行生物信息学分析,并采用实时荧光定量PCR(qPCR)检测GnIH基因在牦牛各器官中的表达情况,通过免疫组织化学和免疫荧光法检测GnIH在组织中的表达及定位。结果表明:GnIH基因编码序列(CDS)区大小为591 bp,编码196个氨基酸;牦牛GnIH基因与黄牛和野牦牛同源性最高;GnIH前体为不稳定亲水酸性蛋白,且无跨膜结构,在第26,27个氨基酸间有信号肽区域,同时GnIH前体包含了3个磷酸化位点,二级结构主要由α-螺旋和无规则卷曲组成。GnIH基因在卵巢中的相对表达量显著高于其他组织(P<0.05);随着卵泡的增长发育,GnIH基因在卵泡颗粒细胞中的表达量呈上升趋势,大卵泡(≥6 mm)中GnIH基因相对表达量最高,显著高于中(>3~<6 mm)、小卵泡(≤3 mm)(P<0.05),而中、小卵泡之间差异不显著(P>0.05)。GnIH在卵巢颗粒细胞的细胞核及细胞质中均有表达。说明牦牛GnIH基因参与了牦牛的卵泡发育调控。In order to explore the sequence characteristics of gonadotropin-inhibitory hormone(GnIH)gene and its expression and localization in yak ovary granule cells,in this experiment,GnIH gene sequence was cloned from cDNA of yak ovary tissue and bioinformatics analysis was carried out using online biological software.Real-time PCR(qPCR)was used to detect the expression of GnIH gene in yak tissues.Expression and localization of GnIH in tissues were detected by immunohistochemistry and immunofluorescence.The results showed that the size of the coding sequence(CDS)region of GnIH gene was 591 bp,encoding 196 amino acids.Yak GnIH gene had the highest homology with cattle and wild yak.GnIH precursor was an unstable hydrophilic acidic protein with no transmembrane structure,signaling peptide region between amino acids 26 and 27;while GnIH precursor contained 3 phosphorylation sites,and the secondary structure was mainly composed ofα-helix and irregular coil.The expression of GnIH gene in ovary was significantly higher than those of other tissues(P<0.05).With the growth and development of follicles,the expression of GnIH gene in follicular granule cells showed an upward trend;the expression of GnIH gene was the highest in large follicles(≥6 mm),which was significantly higher than those of medium(>3-<6 mm)and small follicles(≤3 mm)(P<0.05),while the difference between medium and small follicle groups was not significant(P>0.05).GnIH was expressed in both the nucleus and cytoplasm of ovarian granule cells.The results indicated that yak GnIH gene was involved in the regulation of yak follicle development.

关 键 词：牦牛 GnIH基因 序列特征 颗粒细胞 亚细胞定位 

分 类 号：S823.85[农业科学—畜牧学]