摘要
旨在预测山羊β防御素104a蛋白质特性,研究其在成年公山羊生殖组织及其他组织中的表达特性及其定位。采用在线软件对gBD104a基因(KJ508074.1)进行生物信息学分析,用QRT-PCR法检测gBD104amRNA在睾丸、附睾及其他组织中的表达情况,用免疫荧光技术对生殖组织和精子中的gBD104a进行定位。结果,(1)生物信息学分析结果表明gBD104a基因cDNA全长324bp,编码107个氨基酸,第1~19个氨基酸为信号肽,第27~55个氨基酸为潜在的β防御素构象区域,在C端有7个潜在的O糖基化位点;(2)QRT-PCR结果表明,gBD104a在附睾体中表达量最高,在附睾头、气管、皱胃、空肠、回肠等组织中表达量较高,其他组织中表达量均较低;(3)免疫组化定位结果显示,在附睾头和体部的假复层纤毛柱状上皮细胞检测到较强的gBD104a阳性信号,在附睾尾部的纤毛柱状上皮细胞也检测到较强的阳性信号。gBD104a包裹在精子头部顶体和中段线粒体表面。gBD104a是一种分泌型糖蛋白,在成年公山羊机体各组织中广泛表达,但主要表达在附睾体,是精子表面的主要成分。推测可能与精子运动、顶体反应和获能有关。需要进一步深入研究其功能。
The objectives of this study were to analyze the goat beta-defensins 104a(gBD104a)gene by bioinformatics,and to investigate the expression levels of gBD104 ain various tissues of adult bucks and the location in reproduction organ and sperms.Bioinformatics analysis of gBD104a(GenBank No.:KJ508074.1)was made by software online.Relative mRNA expressions were detected by real-time fluorescent quantitative PCR.Cellular localization of gBD104 ain testes,epididymidis and sperms were examined by immunohistochemistry.The results showed that:(1)The results of bioinformatics analysis showed that the cDNA of gBD104 agene contains324nucleotides,including a complete open reading frame(ORF)encoding 107 amino acids,software online predictded 1-19 amino acids as signal peptide area,27-55 amino acids for potential beta defensins motif,7potential O-glycosylation sites on C-terminal;(2)The results of QRT-PCR showed that gBD104 a mRNA was widely expressed,but the relative expression of gBD104 amRNA was the highest in cauda epididymis.Expressions of gBD104 amRNA in caput epididymis,tracheal,abomasums,jejunum and ileum were more than those in other tissues;(3)The results of immunohistochemistry showed that there were strong positive signal of gBD104 ain pseudostratified ciliated columnar epithelium of caput and corpus,and strong positive signal in columnar epithelium of cauda.gBD104 awas located on the sperm acrosome and mitochondria surface.gBD104 a was an epididymal secretory glycoprotein.gBD104 a mRNA was widely expressed in various tissues of bucks,while the highest expression levels was in cauda epididymis.gBD104 awas coated on sperm surface,which maybe play an important role in sperm motility,acrosome reaction and capacitation process.Therefore,the physiological functions of gBD104 aneeds be further researched.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2015年第2期219-227,共9页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
山西省科技攻关计划项目(20110311029
20130311025-2)
关键词
β防御素104a
生殖组织
表达特性及定位
公山羊
goat beta-defensin 104a
reproductive tissues
expression characteristics and localization
bucks
