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。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。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 神经元活性的其他神经元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。成年恒河猴 (macaca mulatta) 下丘脑中的 LHRH 基因表达受性腺类固醇调节，主要是雌性的雌激素和雄性的雄激素。 尽管存在这种区别，但尚无关于性别特异性信号如何调节 LHRH 基因表达的报道。 从性腺完整的成年雄性（n=5，11.0±1.7岁）和雌性（n=5，11.2±2.0岁）恒河猴的内侧基底下丘脑（MBH）和视前区（POA）收集RNA和DNA。雌性猴子的排卵周期阶段是未知的，因为它们是从组织分配程序中获得的。使用定量PCR，我们发现男性MBH中的LHRH mRNA水平显着低于女性（p0.05），但POA中没有差异。此外，我们发现这两个区域的 GAD65、GAD67、NPY、KiSS-1 和 GPR54 mRNA 不存在性别差异。因为灵长类动物下丘脑 LHRH 神经分泌系统中不存在永久性性别分化，即，切除睾丸的成年雄性猴中的卵巢移植可诱导 LH 激增（Norman and Spies Endocrinology 118：2608-2610，1986），因此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（一种影响转录因子结合和基因表达的 DNA 共价修饰）在性别之间是否存在差异。初步调查显示，在 LHRH 基因 5' 侧翼区域，男性的 CpG 甲基化水平高于女性（p0.1）。该甲基化位于相对于恒河猴 LHRH 基因转录起始位点的 CpG 丰富区域 -2100/-1700。总的来说，这些结果表明恒河猴中 LHRH mRNA 水平存在显着的性别差异，这可能与 CpG 甲基化的差异有关。结论是表观基因组修饰是灵长类 LHRH 基因表达复杂协调的因素。 本研究使用了 WNPRC 动物服务（病理学服务）。