Sperm histone code and epigenetic-mediated subfertility

精子组蛋白密码和表观遗传介导的生育力低下

• 批准号: 45912574
• 负责人: Professor Dr. Klaus Steger
• 金额: --
• 依托单位: Klinik und Poliklinik für Urologie, Kinderurologie und Andrologie
• 依托单位国家: 德国
• 项目类别: Clinical Research Units
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/45912574?language=en
• 关键词: Sperm; histone; code; epigenetic; mediated

It is now generally accepted that a sperm-specific epigenetic code is established during spermiogenesis and transmitted to the oocyte. Aberrations may represent one reason for idiopathic male infertility resulting in decreased IVF/ICSI outcome. In a previous study, gene promoters associated with histone H4 acetylated at lysine 12 (H4K12ac) and also exonic sequences associated with histone H3 acetylated at lysine 9 (H3K9ac) have been identified in ejaculated spermatozoa from fertile donors and subfertile patients applying ChlP-on-chip HG18 promoter array and ENCODE array, respectively. In the present study, the following aims will be addressed: 1) To clarify whether H4K12ac-associated gene promoters colocalize with other histones, such as H3K9ac, H3K4me3 and H3K27me3, or protamine applying Re-ChIP; 2) To determine binding sequences for protamine-1 iri fertile donors and subfertile patients; 3) To analyze the methylation status of CpG dinucleotides within gene promoters of developmental genes associated with H4K12ac and protamine-1 applying pyrosequencing; 4) To analyze the possible role of H4K12ac and H3K9ac on paternal imprinted loci; 5) To analyze expression of H4K12ac-associated genes in the paternal pronucleus of mouse zygote using in-situ hybridization and immunofluorescence in combination with confocal microscopy; 6) To perform a genome-wide bioinformatic-based data comparision of own data from H4K12ac ChlP-on-chip and protamine-1 ChlPsequencing with transcriptomes from human preimplantation embryos/embryonic stem cells and data generated in bovine model providing paternal contributed candidate genes for eariy development (coop, with project 7).

现在人们普遍认为，精子特异性表观遗传密码是在精子发生过程中建立并传递给卵母细胞的。畸变可能是特发性男性不育症导致 IVF/ICSI 结果下降的原因之一。在之前的一项研究中，应用 ChlP-on-chip HG18 启动子阵列和 ENCODE 阵列，在来自生育力供体和生育力低下患者的射精精子中分别鉴定了与赖氨酸 12 乙酰化组蛋白 H4 (H4K12ac) 相关的基因启动子以及与赖氨酸 9 乙酰化组蛋白 H3 (H3K9ac) 相关的外显子序列。在本研究中，将解决以下目标：1）阐明H4K12ac相关基因启动子是否与其他组蛋白（例如H3K9ac、H3K4me3和H3K27me3）或应用Re-ChIP的鱼精蛋白共定位； 2) 确定鱼精蛋白-1 iri 生育力供体和生育力低下患者的结合序列； 3) 应用焦磷酸测序分析与H4K12ac和鱼精蛋白1相关的发育基因的基因启动子内CpG二核苷酸的甲基化状态； 4) 分析H4K12ac和H3K9ac对父本印记位点的可能作用； 5) 采用原位杂交和免疫荧光结合共聚焦显微镜分析小鼠受精卵父本原核中H4K12ac相关基因的表达； 6) 对来自 H4K12ac ChlP-on-chip 和鱼精蛋白-1 ChlP测序的自身数据与来自人类植入前胚胎/胚胎干细胞的转录组和牛模型中生成的数据进行基于全基因组生物信息学的数据比较，为早期发育提供父系贡献的候选基因（合作社，项目7）。