LHR、PRLR基因在不同发育阶段牦牛睾丸中的表达与定位  

Expression and Localization of LHR and PRLR Genes in Testicles of Yak(Bos grunniens) at Different Developmental Stages

作　　者：马富龙 褚敏[1] 常永芳 包鹏甲[1] 郭宪[1] Qudratullah Kalwar[1] 马启财 潘和平 阎萍[1] MA Fu-Long;CHU Min;CHANG Yong-Fang;BAO Peng-Jia;GUO Xian;Qudratullah Kalwar;MAQi-Cai;PAN He-Ping;YAN Ping(Lanzhou Institute of Husbandry and Pharmaceutical Sciences,Chinese Academy of Aicultural Sciences/Gansu Key Laboratory for Yak Breeding Engineering of Gansu Province,Lanzhou 730050,China;College of Life Science and Engineering,Northwest University for Nationlities,Lanzhou 730030,China)

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所/甘肃省牦牛繁育工程重点实验室,兰州730050 [2]西北民族大学生命科学与工程学院,兰州730030

出　　处：《农业生物技术学报》2020年第4期672-680,共9页Journal of Agricultural Biotechnology

基　　金：中国农业科学院创新工程项目(CAAS-ASTIP-2014-LIHPS-01);现代肉牛牦牛产业技术体系(CARS-37)。

摘　　要：促黄体素受体(luteinizing hormone receptor, LHR)和催乳素受体(prolactin receptor, PRLR)基因在哺乳动物生殖发育中扮演着重要角色,广泛存在于各种组织。为分析不同年龄牦牛(Bos grunniens)睾丸组织中LHR、PRLR基因表达的变化模式,并探究其在成年期牦牛不同器官中的表达差异,分别采集幼年期(6月龄)、初情期(24月龄)、性成熟期(36月龄)、成年期(72月龄)牦牛睾丸组织和72月龄公牦牛的肾脏、肝脏、肺脏、心脏、脾脏、骨骼肌等器官组织。采用qRT-PCR技术检测不同发育时期牦牛睾丸组织和成年牦牛不同器官组织中LHR、PRLR mRNA表达水平,并采用免疫组织化学技术研究不同发育期牦牛睾丸组织的LHR、PRLR蛋白表达定位。结果显示,LHR、PRLR基因在牦牛睾丸组织中的表达量从幼年期到性成熟期极显著增加(P<0.01),成年期与性成熟期差异不显著。成年牦牛不同器官组织LHR基因在睾丸组织中的表达水平高于在肾脏、骨骼肌和肺脏组织中的表达水平(P<0.05),PRLR基因在肾脏的表达量最高,其次为骨骼肌、睾丸和脾脏(P<0.05)。LHR、PRLR免疫阳性标记定位于睾丸支持细胞、间质细胞、初级精母细胞和次级精母细胞,其在细胞中的阳性信号从幼年期到性成熟期逐渐增强,成年期趋于稳定,蛋白表达趋势与mRNA基本相符。说明睾丸组织中LHR、PRLR可能在精子成熟过程中发挥作用并参与间质细胞的生物学功能,LHR还可能参与调节其他非性腺组织的发育。上述研究结果为揭示牦牛睾丸发育与生殖调控的分子机制提供重要理论依据。Luteinizing hormone receptor(LHR) and prolactin receptor(PRLR) genes play essential roles in mammalian reproductive development. But still no any available research regarding these genes in yak(Bos grunniens). Therefore this study was planned to explore the role of LHR and PRLR genes in different tissues of Yak during different developmental ages(6, 24, 36 and 72 months). The mRNA expression levels of LHR and PRLR in the testis tissues of yak at different developmental stages and organs of yak were detected by q RTPCR, while the protein expression of LHR and PRLR were evaluated by immunohistochemistry. The results showed that mRNA expression levels of LHR and PRLR genes in yak testis increased significantly from infancy to sexual maturity(P<0.01), but there was no significant difference between adult and sexual maturity. The expression level of LHR gene mRNA in different organs of adult yak was higher in testis than in kidney, skeletal muscle and lung(P<0.05). While the expression level of PRLR mRNA was higher in kidney,followed by skeletal muscle, testis and spleen(P<0.05). Additionally, immunohistochemistry findings revealed that LHR and PRLR proteins were located in testicular supporting cells, mesenchymal cells, primary spermatocytes and secondary spermatocytes, and they were increased gradually from infancy to sexual maturity stage and tended to be stable in adulthood. These findings displayed that LHR and PRLR may be vital for sperm maturation, in the biological functions of interstitial cells, and in regulating the development of other non-gonadal tissues. These results provide an important theoretical basis for revealing the molecular mechanism of testis development and reproductive regulation in yaks.

关 键 词：牦牛 睾丸 促黄体素受体基因 催乳素受体基因 基因表达 蛋白定位 

分 类 号：S823.83[农业科学—畜牧学]