摘要
目的了解人睾丸组织中ACRBP的表达及其启动子CpG位点的甲基化状态,为探讨该基因表达机制奠定基础。方法运用免疫组化法研究睾丸组织中目的蛋白定位;结合生物信息学分析该基因启动子CpG位点序列;通过Bisulfite PCR测序法(BSP)分析目的基因CpG位点的甲基化状态。结果免疫组化显示精子与精子细胞及部分的精原细胞和精母细胞表达目的蛋白。对ACRBP启动子序列(转录起始点-127～+110 bp)进行BSP扩增测序,发现该区域共有24个CpG位点,其中仅有3个出现甲基化,其甲基化频率为1.67%。结论生精小管中ACRBP蛋白呈区域性的阳性反应,可能与生精小管内精子发生的不同步性有关;ACRBP启动子CpG位点低甲基化可能与该基因表达的机制有关。
Objective To investigate the expression of acrosin binding protein（ACRBP）and CpG site methylation of its promoter in human normal testis for conforring expression mechanisms of ACRBP.Methods ACRBP expression in human testis was detected by immunohistochemical staining with polyclonal antibody;the proximal promoter of ACRBP was bioformatically analyzed to obtain its sequence;the methylation of CpG site was analyzed by bisulfite sequencing PCR（BSP）.Results Seminiferous tubles had strong intratubular staining of mostly spermatozoon and spermatid with less staining of spermatogonium and spermatocyte by ACRBP polyclonal antibody.Three of 24 CpG sites were methylated during the DNA fragment（-127-＋110 bp） of ACRBP gene amplified,and the methylation frequence was 1.67%.Conclusion Asynchronous spermatogenesis may lead to the reginal location of ACRBP in seminiferous tubles;the low CpG site methylation of ACRBP maybe related to its mechanism of expression.
出处
《山东医药》
CAS
北大核心
2011年第47期21-23,共3页
Shandong Medical Journal
基金
国家自然科学基金项目(30760055)
广西高发疾病研究创新团队基金项目(桂教人2007-59号)
广西教育厅立项项目(200911LX2022
01106LX265)
广西中医药管理局立项课题项目(gzzc0902)
