PTIP as an epigenetic regulator in male meiosis

PTIP 作为雄性减数分裂的表观遗传调节剂

• 批准号: 8672663
• 负责人: GUO-ZHANG ZHU
• 金额: $ 7.23万
• 依托单位: MARSHALL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-05 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8672663
• 关键词: Address; Architecture; Cell Differentiation process; Chromatin; Chromosomes; Complex; Comprehension; Contraceptive Agents; DNA Methylation; Data; Development; Embryonic Development; Epigenetic Process; Event; Functional RNA; Genetic; Genetic Variation; Genome; Germ Cells; Goals; Histone H3; Human; Immunofluorescence Immunologic; Infertility; Knock-out; Lead; Link; Lysine; Male Contraceptions; Mass Spectrum Analysis; Meiosis; Meiotic Prophase I; Meiotic Recombination; Methylation; Methyltransferase; Molecular; Mus; Outcome; Phenotype; Play; Process; Proteins; Regulation; Reproduction; Research; Role; Somatic Cell; Spermatocytes; Spermatogenesis; Sterility; Testing; Testis; Time; Transcriptional Activation; Transgenic Mice; Work; assisted reproduction; chromatin remodeling; epigenome; histone modification; immunoaffinity chromatography; insight; male; men; mouse model; novel; novel strategies; paternal imprint; postnatal; public health relevance

DESCRIPTION (provided by applicant): It is becoming evident that epigenetic mechanisms, including DNA methylation, histone modification and non-coding RNAs, play significant roles in spermatogenesis. Despite recent progress, our comprehension of the dynamic epigenome in developing male germ cells remains largely to be defined. Thus, it is time to identify novel epigenetic regulators and discover how they work in spermatogenesis. The goal of this application is to test the hypothesis that PTIP (PAX transcription activation domain interacting protein) is an essential epigenetic regulator in male meiosis. PTIP has been lately identified as part of a histone H3 lysine 4 (H3K4) methyltransferase complex and is essential for embryonic development. H3K4 methylation is an epigenetic mark and is involved in the regulation of genome function through modifying chromatin architecture. Given dynamic chromatin remodeling in spermatogenesis, we recently generated a transgenic mouse model with conditional knockout of PTIP in postnatal male germ cells. Remarkably, our preliminary data reveal that conditional knockout of PTIP in male germ cells leads to spermatogenesis arrest in meiosis prophase I, small testes, azoospermia, and sterility. Accordingly, two Specific Aims will be pursued in this application to test our hypothesis. Specific Aim 1: Further analyze meiotic phenotypes of PTIP- deficient male germ cells. We will accomplish this aim by implementing meiotic chromosome spread and immunofluorescence. Specific Aim 2: Determine PTIP-interacting proteins in spermatocytes. We will employ immunoaffinity chromatography followed by mass spectrometry to identify PTIP's functional partners. Successful completion of this project will considerably further our understanding of the germ cell epigenome during spermatogenesis and broadly define an epigenetic mechanism of cell differentiation and development. Furthermore, collected information may offer insights into unexplained (or idiopathic) infertility in men and may lead to new strategies for male contraception and assisted reproduction.

描述（由申请人提供）：越来越明显的是，表观遗传机制，包括DNA甲基化，组蛋白修饰和非编码RNA，在精子发生中发挥重要作用。尽管最近的进展，我们的理解动态表观基因组在发展中的男性生殖细胞仍然在很大程度上有待确定。因此，现在是时候确定新的表观遗传调节因子，并发现它们如何在精子发生中发挥作用。本申请的目的是检验PTIP（PAX转录激活结构域相互作用蛋白）是雄性减数分裂中必需的表观遗传调节因子的假设。PTIP最近被鉴定为组蛋白H3赖氨酸4（H3K4）甲基转移酶复合物的一部分，对胚胎发育至关重要。H3K4甲基化是一种表观遗传标记，通过修饰染色质结构参与基因组功能的调节。鉴于动态染色质重塑精子发生，我们最近产生了一个转基因小鼠模型与条件敲除PTIP在出生后的男性生殖细胞。值得注意的是，我们的初步数据显示，PTIP在男性生殖细胞中的条件性敲除导致精子发生停滞在减数分裂前期I，小睾丸，无精子症和不育。因此，在本申请中将追求两个特定目的来测试我们的假设。具体目的1：进一步分析PTIP缺陷型雄性生殖细胞的减数分裂表型.我们将通过实施减数分裂染色体扩散和免疫荧光来实现这一目标。具体目标2：确定精母细胞中PTIP相互作用蛋白。我们将采用免疫亲和层析，然后质谱法，以确定PTIP的功能合作伙伴。该项目的成功完成将大大加深我们对精子发生过程中生殖细胞表观基因组的理解，并广泛定义细胞分化和发育的表观遗传机制。此外，收集的信息可能会提供对男性不明原因（或特发性）不育症的见解，并可能导致男性避孕和辅助生殖的新策略。