畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (8): 1642-1650.doi: 10.11843/j.issn.0366-6964.2018.08.009

• 生物技术与繁殖 • 上一篇    下一篇

不同发育阶段湖羊生殖器官脂联素受体与睾酮分泌关键基因表达模式及相关性研究

姚晓磊1,2, 黄欣爱3, 肖慎华1,2, 郑临枫1, 范丽洁1, 金宇月1, 刘孜斐1,2, 张艳丽1,2, 王洁3, 王锋1,2*   

  1. 1. 南京农业大学动物科技学院, 南京 210095;
    2. 南京农业大学羊业科学研究所, 南京 210095;
    3. 江苏农牧科技职业学院, 泰州 225300
  • 收稿日期:2018-01-29 出版日期:2018-08-23 发布日期:2018-08-23
  • 通讯作者: 王锋,E-mail:caeet@njau.edu.cn
  • 作者简介:姚晓磊(1989-),男,山西运城人,博士,主要从事动物遗传育种与繁殖研究,E-mail:2016205004@njau.edu.cn
  • 基金资助:

    国家科技支撑计划(2015BAD03B05-06);江苏省重点研发计划(BE2015362);江苏省研究生科技创新项目(KYCX17_0608)

Expression Patterns and Correlation of Adiponectin Receptors and Testosterone Secretion Related-genes Expression in Male Reproductive Organs at Different Developmental Stages of Hu Sheep

YAO Xiao-lei1,2, HUANG Xin-ai3, XIAO Shen-hua1,2, ZHENG Lin-feng1, FAN Li-jie1, JIN Yu-yue1, LIU Zi-fei1,2, ZHANG Yan-li1,2, WANG Jie3, WANG Feng1,2*   

  1. 1. College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China;
    2. Institute of Sheep Industry Science, Nanjing Agricultural University, Nanjing 210095, China;
    3. Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China
  • Received:2018-01-29 Online:2018-08-23 Published:2018-08-23

摘要:

旨在探究不同发育阶段湖羊生殖器官中脂联素受体(adiponectin receptor,AdpR)及睾酮(testosterone,T)分泌相关基因mRNA的表达变化及相关性分析。选取3、9、24月龄(M)湖羊各5只,颈静脉采血并收集血清,屠宰后,采集下丘脑、垂体、睾丸、附睾头、体、尾组织;ELISA技术检测各月龄湖羊血清中脂联素(adiponectin,Adp)、T、促性腺激素释放激素(gonadotropin releasing hormone,GnRH)、促性腺激素抑制激素(gonadotropin inhibitory hormone,GnIH)浓度;qRT-PCT技术检测不同发育阶段湖羊下丘脑-垂体-性腺轴(hypothalamus-pituitary-gonad axis,HPG)中AdpR1和AdpR2及T分泌关键基因mRNA表达规律;免疫组化技术对AdpR1在湖羊睾丸和附睾中进行定位分析。结果表明:不同发育阶段湖羊血清中Adp、GnRH、GnIH浓度呈现不同的变化趋势,其中Adp含量在性成熟后(9和24月龄)显著高于性成熟前(3月龄)(P<0.05)。AdpR1 mRNA表达量在各个月龄睾丸和附睾尾中均显著高于AdpR2(P<0.05),其中AdpR1 mRNA表达量在二者中呈上升趋势,且差异显著(P<0.05),而在附睾头、体差异不显著(P>0.05)。AdpR1存在于睾丸间质细胞、生精细胞、曲精细管管壁肌样细胞、精子细胞以及附睾头、体、尾的上皮细胞。血清中Adp与T含量呈显著正相关(P<0.05);Adp含量与GnIH负相关以及与GnRH正相关,但差异不显著(P>0.05);AdpR1 mRNA表达水平与StAR、3β-HSD mRNA表达水平呈显著正相关(P<0.05)。综上表明,在湖羊生殖器官发育过程中,Adp通过与其受体AdpR1结合,促进T的产生,进而影响湖羊睾丸发育。

Abstract:

The aims of this study were to observe the expression patterns and the correlation of adiponectin receptors (AdpR) and testosterone (T) synthesis-related genes in male reproductive organs at different developmental stages of Hu sheep. Five Hu sheep at ages of 3, 9, 24 month (M) were randomly selected, respectively. Blood was collected from the jugular vein of each ram to separate the plasma. Tissue samples including hypothalamus, pituitary, testis, caput, corpus and cauda were collected from the different developmental stages of Hu sheep after the slaughter. The concentrations of serum adiponectin (Adp), T, gonadotropin releasing hormone (GnRH) and gonadotropin inhibitory hormone (GnIH) at different ages were analyzed by ELISA, and the expression patterns of AdpR1, AdpR2 and T synthesis-related genes mRNA in male reproductive tract at the 3 developmental stages of Hu sheep were examined by qRT-PCR. The location of AdpR1 in the male reproductive tract was detected by immunohistochemical. The results showed that the levels of Adp, GnRH and GnIH in serum had different change trends. Adp concentration were significantly higher in adult (9 and 24 month) group than pre-pubertal (3 month) group. The mRNA expression of AdpR1 in testis and epididymis cauda at different developmental stages was significantly higher than that of AdpR2 (P<0.05). Moreover, the mRNA expression of AdpR1 in the testis and cauda increased significantly with the increasing age (P<0.05), but no significant difference between epididymis caput and corpus (P>0.05). AdpR1 expressed in the ram testis and epididymis, and was mainly localized in leydig cells, spermatocytes, peritubular myoid cells and principal cells. Furthermore, Adp and T levels in serum had significant positive correlation (P<0.05). Although the content of Adp was negatively correlated with GnIH levels and positively correlated with GnRH, respectively, there was no significant difference among them (P>0.05). Meanwhile, the mRNA expression of AdpR1 were significantly positive correlated with StAR and 3β-HSD, respectively (P<0.05). In conclusion, during the development of the reproductive organs of Hu sheep,Adp may promote production of T by combining AdpR1, and then promote the testis development of Hu sheep.

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