瘦素及其受体在牦牛附睾的时空表达特性
Temporal and Spatial Expression Characteristics of Leptin and Its Receptor in Yak(Bos grunniens)Epididymis
王晓艳 1崔燕 2余四九 2马睿 1何俊峰 1张虔 1龙敏1
作者信息
- 1. 甘肃农业大学动物医学院,兰州 730070
- 2. 甘肃农业大学动物医学院,兰州 730070;甘肃省牛羊胚胎工程技术研究中心,兰州 730070
- 折叠
摘要
瘦素(leptin)及瘦素受体(leptin receptor,LEPR)在雄性动物生殖调控中发挥重要作用.为探究Leptin及LEPR在性成熟前后牦牛(Bos grunniens)附睾头、体、尾中的分布特点及表达动态,本研究采集性成熟前(幼年)及性成熟后(成年,老年)牦牛附睾不同部位(头,体,尾),利用qPCR、Western blot和免疫组织化学染色方法检测Leptin及LEPR在不同年龄牦牛附睾头、体、尾中的分布及表达水平.免疫组化结果显示,Leptin和LEPR主要表达在附睾上皮、基底细胞和平滑肌细胞,其中附睾上皮呈强阳性表达;qPCR结果显示,Leptin及LEPR在幼年牦牛附睾头、体和尾的基因表达量显著高于成年和老年(P<0.05),同时发现,Leptin及LEPR在幼年附睾尾表达量显著高于附睾头和附睾体(P<0.05),瘦素及其受体在幼年牦牛的附睾头和附睾体中差异不显著(P>0.05);Western blot结果显示,Leptin及LEPR蛋白在不同年龄牦牛附睾中均有表达,其表达量在幼年组最高,从幼年、成年及老年,依次呈递减趋势,且各年龄之间差异显著(P<0.05);同时发现,Leptin及LEPR蛋白在幼年附睾尾部表达量最高,两种蛋白在幼年附睾头与附睾体之间差异不显著(P>0.05);Leptin及LEPR蛋白在成年和老年附睾中的表达量为附睾尾>附睾头>附睾体,且各部位之间差异显著(P<0.05).结果表明,牦牛性成熟前后附睾中的Leptin及LEPR表达量发生显著变化,并且性成熟后维持在一定水平持续表达至老年,在附睾不同区段的表达也存在差异,附睾尾表达相对最高,说明尾部环境具有较高的Leptin浓度.Leptin及LEPR在牦牛附睾中的时空表达差异提示二者对于牦牛精子的成熟、存储和能量保存方面具有重要意义,本研究为探究高原环境下雄性牦牛的生殖调控机制提供研究资料.
Abstract
Leptin and leptin receptor(LEPR)play important roles in the regulation of male reproduction.This study was conducted to investigate the distribution characteristics and possible roles of Leptin and LEPR in the caput,corpus,and cauda of yak(Bos grunniens)epididymis.Epididymal tissues of yaks of different ages were collected to detect exact distribution and expression levels of Leptin and LEPR in the caput,corpus,and cauda of epididymis using qPCR,Western blot and immunohistochemistry techniques.Immunohistochemical results showed Leptin and LEPR mainly expressed in the epididymal epithelial cells,basal cells,and smooth muscle cells.qPCR results showed that the gene expression levels of Leptin and LEPR in the caput,corpus and cauda of the epididymis of juvenile yak were significantly higher than those of adult and eged yak(P<0.05),and the expression levels of Leptin and LEPR in the cauda of the juvenile yak were significantly higher than those in the caput and corpus of the epididymis of juvenile yak(P<0.05)and there was no significant difference in the caput and corpus of the epididymis of juvenile yak(P>0.05);Leptin and LEPR were not significantly different in the caput,corpus and cauda of the epididymis of adult and aged yaks(P>0.05).Western blot results showed that Leptin and LEPR were expressed in the epididymis of yaks at different ages,and the expression levels of Leptin and LEPR were the highest in juvenile yaks,and decreased successively from juvenile,adult and aged yaks.Western blot results showed that Leptin and LEPR proteins were attached in yaks at different ages.The expression level was the highest in the young group,and showed a decreasing trend from juvenile,adult and old age,with significant differences among different ages(P<0.05);Leptin and LEPR proteins were found to have the highest expression levels in the cauda of juvenile epididymis.There was no significant difference between the caput of the epididymis and the corpus of the epididymis(P>0.05),but there was significant difference between the cauda of the epididymis and the two parts of the epididymis(P<0.05).At the same time,the expression levels of Leptin and LEPR proteins were the highest in the cauda of juvenile epididymis,and there was no significant difference between the two proteins in the caput of juvenile epididymis and the corpus of juvenile epididymis(P>0.05).The expression levels of Leptin and LEPR proteins in adult and aged epididymis were cauda>caput>corpus,and there were significant differences among different parts(P<0.05).The expression levels of Leptin and LEPR were also different in different parts of the epididymis,and the caudal epididymis was relatively the highest,indicating that the caudal environment had a high concentration of Leptin.The temporal and spatial expression differences of Leptin and LEPR in yak epididymis suggest that Leptin and LEPR play important roles in the maturation,storage and energy conservation of yak sperm.This study provides certain research data for the reproductive regulation mechanism of male yaks in plateau environment.
关键词
牦牛/附睾/瘦素(leptin)/瘦素受体(LEPR)Key words
Yak/Epididymis/Leptin/Leptin receptor(LEPR)引用本文复制引用
出版年
2024