Regulation of chromatin remodeling during spermiogenesis

精子发生过程中染色质重塑的调节

基本信息

批准号：
8815702
负责人：
PRABHAKARA P REDDI
金额：
$ 7.86万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-12-01 至 2015-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8815702
关键词：
Accounting Acrosome Adult Affect Applications Grants Archives Assisted Reproductive Technology Binding Biological Models Birth Breeding Bypass CCCTC-binding factor Cell Nucleus Cells ChIP-seq Chromatin Complex Conceptions Congenital Abnormality Couples DNA-Binding Proteins Defect Developed Countries Development Disease Embryonic Development Epigenetic Process European Event Feasibility Studies Fertility Fertilization Fetal Growth Retardation Flagella Genes Genetic Genetic Transcription Genome Germ Cells Germ Lines Histones Human Immunohistochemistry Infertility Injection of therapeutic agent Intercistronic Region Knock-out Knockout Mice Lead Life Male Infertility Malignant Childhood Neoplasm Methods Modeling Molecular Mus Mutant Strains Mice Natural Selections Nuclear Nuclear Proteins Phase Physical condensation Physical shape Pilot Projects Play Preparation Process Protamines Proteins Regulation Reproductive Health Risk Role Sperm Maturation Spermatids Spermatogenesis Spermiogenesis Spontaneous abortion Staging Technology Testing Testis Time Treatment Factor United States Ursidae Family Validation Work Zinc Fingers age group base chromatin remodeling egg embryonic stem cell gene repression genome-wide imprint improved insight knockout gene male mouse model novel promoter public health relevance reproductive research study screening sperm cell technology development transcription factor

项目摘要

DESCRIPTION (provided by applicant): The differentiation of round spermatids into spermatozoa, known as spermiogenesis, is a complex process involving the formation of the acrosome, the flagellum, and condensation of the nucleus. In preparation for condensation, the spermatid nucleus undergoes dramatic chromatin remodeling including genome-wide cessation of transcription, dismantling of the nucleosomal organization, and histone to protamine transition. This process is unique to the male germ line. Mouse models have shown that defects in nuclear condensation lead to male infertility; however, the mechanisms regulating this complex genome-wide process are not well understood. Our working hypothesis is that the genome organizer protein CTCF coordinates the chromatin remodeling events accompanying spermatid differentiation. The evolutionarily conserved eleven zinc finger protein CCCTC-binding factor (CTCF) is exclusively expressed within the round and early elongating spermatids in mice, coinciding with transcriptional shutdown and the onset of histone replacement. In this pilot, feasibility grant proposal we will test the hypotheses that: 1) CTCF functions as a transcription factor as well as an organizer of spermatid genome to facilitate chromatin remodeling and 2) CTCF is essential for the completion of spermatogenesis and male fertility. We will perform ChIP-seq to determine genome-wide occupancy of CTCF in round spermatids and generate CTCF conditional knockout mice using the Cre-loxP technology to test the requirement of CTCF for spermatogenesis and male fertility. CTCF is an ideal candidate for this role because it is a multifunctional DNA binding protein with diverse roles including that of a chromatin organizer. Floxed CTCF mice as well as the male germ cell-specific cre-deleter strain (Stra8-iCre) are readily available. This proposal will explore the role of CTCF in male fertility fr the first time. Infertility affects 1 in 6 couples in the reproductive age group, with the male facor accounting for 50% of those cases. If depletion of CTCF causes sperm maturation arrest, this study will provide a novel mouse model for male infertility. Given the role that CTCF plays in establishing and / or maintaining epigenetic marks, the proposed knockout mouse may be a useful model to understand the risks involved in Assisted Reproductive Technology using incompletely developed spermatids or sperm. Thus, the studies are highly significant from the point of view of male reproductive health.

描述（由应用提供）：将圆形精子分化为精子，被称为精子发生，是一个复杂的过程，涉及跨，鞭毛的形成和核的凝结。为了准备凝结，精子核心经历了戏剧性的染色质重塑，包括转录的全基因组停止，核素组织的拆卸以及蛋白质过渡的组蛋白。这个过程是男性种系独有的。小鼠模型表明，核冷凝的缺陷导致男性不育症。但是，尚不清楚应对这一复杂基因组过程的机制。我们的工作假设是基因组组织者蛋白CTCF协调染色质重塑事件参与精子分化。进化保守的11个锌指蛋白CCCTC结合因子（CTCF）仅在小鼠的圆形和早期伸长的精子中表达，与转录关闭和组蛋白替代的发作一致。在此试验中，可行性赠款提案我们将测试以下假设：1）CTCF充当转录因子以及精子基因组的组织者，以促进染色质重塑和2）CTCF对于完成精子发生和男性生育力至关重要。我们将执行CHIP-SEQ，以确定CTCF在圆形精子中的全基因组占用率，并使用CRE-LOXP技术生成CTCF条件敲除小鼠，以测试CTCF对精子发生和男性生育能力的需求。 CTCF是该角色的理想候选者，因为它是一种多功能DNA结合蛋白，具有染色质组织者（包括染色质组织者）的作用。 Floxed CTCF小鼠以及雄性生殖细胞特异性Cre-deleter菌株（Stra8-Icre）很容易获得。该提案将首次探讨CTCF在男性生育中的作用。不育会影响生殖年龄组的六对夫妇中的1个，而男性面部占50％。如果CTCF的耗竭会导致精子成熟，则该研究将为男性不育症提供新型的小鼠模型。鉴于CTCF在建立和 /或维持表观遗传标记中所扮演的角色，拟议的基因敲除小鼠可能是使用未完全开发的精子或精子来了解辅助生殖技术所涉及的风险的有用模型。从男性复制健康的角度来看，这些研究非常重要。