The role of transcription factor S-SOX5 in male fertility and sperm flagella formation

转录因子S-SOX5在男性生育力和精子鞭毛形成中的作用

• 批准号: 9225891
• 负责人: Zhibing Zhang
• 金额: $ 7.63万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9225891
• 关键词: Address; Binding Sites; Biological Assay; Blood Circulation; Brain; Bronchiectasis; Cartilage; Cell Nucleus; Cerebral Ventricles; Cerebrospinal Fluid; Chondrogenesis; Chromatin; Cilia; Clinical Medicine; Code; Contraceptive methods; Defect; Development; Disease; Drug or chemical Tissue Distribution; Embryonic Development; Enterobacteria phage P1 Cre recombinase; Epithelial Cells; Event; Exons; Fertility; Flagella; Foundations; Gene Expression; Gene Expression Regulation; Gene Family; Gene Targeting; Generations; Genes; Genetic; Genetic Transcription; Germ Cells; Haploidy; Heart; Hydrocephalus; Impairment; Indirect Immunofluorescence; Infertility; Kidney; Knowledge; Length; LoxP-flanked allele; Lung; Male Infertility; Mammalian Oviducts; Meiosis; Messenger RNA; Microtubules; Mus; Mutant Strains Mice; N-terminal; Names; Neural Crest; Neurons; Nuclear; Phase; Phenotype; Play; Process; Promoter Regions; Protein Isoforms; Proteins; Research; Respiratory System; Respiratory tract structure; Role; SOX5 gene; Shapes; Sinusitis; Situs Inversus; Skeletal Muscle; Sperm Maturation; Sperm Motility; Spermatids; Spermatocytes; Spermatogenesis; Spermiogenesis; Stem cells; Structure; Testing; Testis; Time; Tissues; Trachea; Transcript; Transcriptional Regulation; Transgenic Mice; Translating; base; blastomere structure; cell motility; chromatin immunoprecipitation; cilium motility; combinatorial; in vivo; lung development; male; member; novel; particle; pathogen; programs; sperm cell; sperm function; tool; transcription factor; transcriptome sequencing

Motile cilia are present in the epithelial cells of brain ventricles, trachea, oviduct, and sperm flagella. They play important roles in vivo including maintaining cerebrospinal fluid (CSF) circulation, clearing particles and pathogens from the respiratory tract, and sperm motility. Defects in motile cilia result in multiple diseases such as hydrocephalus, situs inversus, male infertility and et al. Most motile cilia have a “9+2” axoneme. The shared structure of motile cilia in different tissues suggests that common regulatory mechanisms exist to govern expression of cilia/flagellar components. We hypothesize that expression of the genes essential for motile cilia structure/function are regulated by similar mechanisms so that the proteins are assembled into the motile cilia in a coordinated and timely manner. Sperm flagella are special motile cilia. Besides the core axoneme, other accessory structures are also assembled during spermiogenesis, the process of spermatid differentiation. During this phase, a number of genes essential for sperm flagella assembly are up-regulated. However, the transcriptional regulation of this suite of genes is poorly understood. By analyzing putative transcription factor binding sites in the promoter regions of genes that are essential for sperm flagella structure/function, we identified SOX5 as a putative key controller of cilia/flagellar genes. Sox-5 is a member of a family of genes that shows homology to the high motility group (HMG) box region of the testis determining gene, SRY. The mouse Sox5 gene encodes two major transcripts: the longer Sox5 isoform, originally named L-Sox5, but most authors refer it as Sox5; and a shorter isoform, S-Sox5. The first exon of the S-Sox5 transcript is a non-translated exon;, and it is not present in the L-Sox5 transcript. The translated 48 kDa S-SOX5 protein lacks the N-terminal half of the 84 kDa L-SOX5 protein. The two SOX5 proteins have different tissue distributions. L-SOX5 is expressed in multiple tissues, including the cartilage, heart, brain, kidney, lung, and skeletal muscle, but not in the testis, and it plays important roles in regulating processes of embryonic development and cell fate determination. S-SOX5 protein is only expressed in tissues with motile cilia, including the brain, lungs, and particularly in the testis. In the testis, it is localized in the nuclei of post-meiotic round spermatids. It has been shown that S-SOX5 regulates expression of several genes highly expressed in the testis. We hypothesize that S-SOX5 regulates expression of a suite of genes that are essential for sperm flagella formation and function. To test this hypothesis, we propose the following Specific Aims:1. To establish the critical the role of S-SOX5 in flagellogenesis and male fertility; and 2. To establish the target genes regulated by S-SOX5 in male germ cells. The proposed studies will help clarify the combinatorial transcription factor code that governs the differentiation and structural transformation of mammalian male germ cells. The tools that will be developed in pursuit of the proposed research will allow us to expand the project to a RO1 application to elucidate the transcriptional mechanisms that control mammalian sperm maturation.

纤毛纤毛存在于脑室，气管，输卵管和精子鞭毛的上皮细胞中。他们玩 体内重要作用，包括维持脑脊液（CSF）循环，清除颗粒和 来自呼吸道和精子运动的病原体。纤毛母亲的缺陷导致多种疾病此类疾病 作为脑积水，现场倒数，男性不育症等。大多数纤毛具有“ 9+2”轴突。共享 纤毛母亲在不同组织中的结构表明，存在常见的调节机制 纤毛/鞭毛成分的表达。我们假设对纤毛母亲必不可少的基因表达 结构/功能由相似的机制调节，以便将蛋白质组装到母纤毛中 以一种协调和及时的方式。精子鞭毛是特殊的主题纤毛。除了核心Axoneme，其他 在精子发生过程中，还组装了辅助结构，即精子分化的过程。 在此阶段，许多对于精子鞭毛组件必不可少的基因被上调。但是， 对这套基因的转录调节知之甚少。通过分析推定的转录因子 基因启动子区域的结合位点对于精子鞭毛结构/功能至关重要，我们 鉴定为Sox5是纤毛/鞭毛基因的推定键控制器。 Sox-5是一个基因家族的成员 显示了睾丸确定基因的高运动组（HMG）框区域的同源性。鼠标 SOX5基因编码两个主要的成绩单：SOX5同工型，最初称为L-SOX5，但大多数作者 将其称为Sox5;和较短的同工型S-SOX5。 S-SOX5转录本的第一个外显子是未翻译的 外显子;并且它不存在于L-SOX5转录本中。翻译的48 kDa S-SOX5蛋白缺乏N末端 84 kDa L-SOX5蛋白的一半。两种SOX5蛋白具有不同的组织分布。 L-SOX5是 在多个时间安排中表达，包括软骨，心脏，大脑，肾脏，肺和骨骼肌肉，但不在 睾丸及其在确定胚胎发育和细胞命运过程中起着重要作用 决心。 S-SOX5蛋白仅在有纤毛的组织中表达，包括大脑，肺和 在睾丸中，它定位在寿命后圆形精子的核中。它一直 表明S-SOX5调节睾丸中高度表达的几种基因的表达。我们假设这一点 S-SOX5调节了精子形成和功能所必需的一组基因的表达。 为了检验这一假设，我们提出以下特定目的：1。确定s-sox5在 鞭毛生成和男性生育能力；和2。建立在男性生殖细胞中由S-SOX5调节的靶基因。 拟议的研究将有助于阐明控制差异化的组合转录因子代码 和哺乳动物雄性生殖细胞的结构转化。追求将开发的工具 拟议的研究将使我们能够将项目扩展到RO1应用程序以阐明转录 控制哺乳动物精子成熟的机制。