EPIGENOMIC RELATIONSHIP TO A SEX DIFFERENCE IN HYPOTHALAMIC LHRH MRNA EXPRESSION

下丘脑 LHRH mRNA 表达性别差异的表观基因组关系

• 批准号: 8173164
• 负责人: Ei Terasawa-Grilley
• 金额: $ 3.1万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173164
• 关键词: 5' Flanking Region; Adult; Age-Years; Androgens; Animals; Area; Binding; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA; DNA Modification Process; Drug or chemical Tissue Distribution; Endocrinology; Epigenetic Process; Estrogens; Exhibits; Female; Funding; Gene Expression; Genes; Gonadal Steroid Hormones; Gonadotropin Hormone Releasing Hormone; Grant; Hypothalamic structure; Institution; Investigation; KISS1 gene; KISS1R gene; Macaca mulatta; Medial; Messenger RNA; Methylation; Modification; Monkeys; Neurons; Neurosecretory Systems; Ovarian; Pathology; Phase; Preoptic Areas; Primates; RNA; Relative (related person); Reporting; Research; Research Personnel; Resources; Services; Sex Characteristics; Signal Transduction; Source; System; Transcription Initiation Site; Transplantation; United States National Institutes of Health; epigenomics; male; programs; reproductive function; research study; sex; transcription factor; trend

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Objective: To investigate whether sex differences exist in gene expression of LHRH as well as other neuronal systems known to influence LHRH neuronal activity. Luteinizing-hormone releasing hormone (LHRH) is a key regulator of reproductive function. Adult rhesus monkey (macaca mulatta) LHRH gene expression in the hypothalamus is regulated by gonadal steroids, primarily estrogens in females and androgens in males. Despite this distinction, there is no report of how sex-specific signals regulate LHRH gene expression. RNA and DNA were collected from the medial basal hypothalamus (MBH) and preoptic area (POA) of gonadally intact adult male (n=5, 11.0¿1.7 years of age) and female (n=5, 11.2¿2.0 years of age) rhesus monkeys. The phases of ovulatory cycle in the female monkeys were unknown, as they were obtained from the tissue distribution program. Using quantitative PCR, we found that LHRH mRNA levels in the MBH are significantly lower in males compared to females (p0.05), but there was no difference in the POA. Moreover, we found that there were no sex differences in GAD65, GAD67, NPY, KiSS-1, and GPR54 mRNA in either area. Because there is no permanent sexual differentiation in the primate hypothalamic LHRH neurosecretory system, i.e., LH surges could be induced by ovarian transplantation in orchidectomized adult male monkeys (Norman and Spies Endocrinology 118:2608-2610, 1986), it is possible that an epigenetic mechanism regulates the sex difference in LHRH gene expression. In the second experiment we investigated if CpG methylation, a covalent modification of DNA that influences transcription factor binding and gene expression, was different between the sexes. Preliminary investigations revealed a trend (p0.1) with males exhibiting higher CpG methylation than females in the 5' flanking region of the LHRH gene. This methylation is in a CpG rich region -2100/-1700 relative to the transcription start site in the rhesus LHRH gene. Collectively, these results indicate that there is a significant sex difference in LHRH mRNA levels in rhesus monkeys and that this may be related to a difference in CpG methylation. It is concluded that epigenomic modifications are factors in the complex orchestration of LHRH gene expression in primates. This research used WNPRC Animal Services (Pathology Services).

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以出现在其他 CRISP 条目中 列出的机构是。 对于中心来说，它不一定是研究者的机构。 目的：探讨 LHRH 以及已知影响 LHRH 神经元活动的其他神经系统的基因表达是否存在性别差异。 黄体生成素释放激素 (LHRH) 是生殖功能的关键调节因子，下丘脑中的 LHRH 基因表达受性腺类固醇（雌性主要是雌激素，雄性主要是雄激素）调节。没有关于性别特异性信号如何调节 LHRH 基因表达的报道。 从性腺完整的成年男性（n=5，11.0±1.7岁）和女性（n=5，11.2±2.0岁）的内侧基底下丘脑（MBH）和视前区（POA）收集RNA和DNA雌性猴子的排卵周期阶段未知，因为它们是通过定量 PCR 获得的，我们发现 LHRH mRNA。与女性相比，男性的 MBH 水平显着较低（p0.05），但 POA 没有差异。此外，我们发现 GAD65、GAD67、NPY、KiSS-1 和 GPR54 没有性别差异。因为灵长类动物下丘脑 LHRH 神经分泌系统中没有永久性的性别分化，即睾丸切除术后的卵巢移植可引起 LH 激增。在成年雄性猴子中（Norman and Spies Endocrinology 118:2608-2610, 1986），表观遗传机制可能调节 LHRH 基因表达的性别差异。在第二个实验中，我们研究了 CpG 甲基化（一种影响 DNA 的共价修饰）是否存在。转录因子结合和基因表达在性别之间存在差异，初步研究显示男性表现出较高的 CpG 甲基化趋势 (p0.1)。相对于恒河猴 LHRH 基因的转录起始位点，该甲基化位于富含 CpG 的区域 -2100/-1700，总体而言，这些结果表明存在显着的性别差异。恒河猴中 LHRH mRNA 水平的变化，这可能与 CpG 甲基化的差异有关。得出的结论是，表观基因组修饰是 LHRH 基因复杂编排的因素。在灵长类动物中的表达。 本研究使用了 WNPRC 动物服务（病理学服务）。