FUNCTIONAL ANALYSIS OF AGE-SPECIFIC FSH ANALOGS USING GENETICALLY ALTERED MICE

使用基因改造小鼠对年龄特异性 FSH 类似物进行功能分析

• 批准号: 7651599
• 负责人: T. RAJENDRA KUMAR
• 金额: $ 22.68万
• 依托单位: WICHITA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-15 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7651599
• 关键词: Accounting; Activins; Affect; Affinity; Age; Aging; Aromatase; Binding; Biochemical; Biological; Biological Process; Biological Testing; Charge; Chorionic Gonadotropin; Clinical; Drug Formulations; Elderly woman; Estrogens; Estrous Cycle; Estrus; Female; Fertility; Follicle Stimulating Hormone; Follicle Stimulating Hormone Receptor; Future; Gene Expression; Genetic; Genetic Models; Genetically Engineered Mouse; Genitourinary system; Glycoproteins; Goals; Gonadal Steroid Hormones; Gonadotropin Hormone Releasing Hormone; Half-Life; Homeostasis; Hormones; Human; Human Follicle Stimulating Hormone; Impairment; Knock-out; Knockout Mice; Luteinizing Hormone; Menstrual cycle; Methods; Monkeys; Morbidity - disease rate; Mus; Oocytes; Outcome; Ovarian; Ovarian Cycles; Ovarian Follicle; Ovarian Stimulations; Ovary; Peptides; Perimenopause; Personal Communication; Phenotype; Physiological; Physiology; Pituitary Gland; Pituitary Gonadotropins; Placenta; Play; Positioning Attribute; Postmenopause; Preparation; Production; Rattus; Regulation; Relative (related person); Research; Role; Serum; Switch Genes; Symptoms; System; Testing; Tetanus Helper Peptide; Tetracyclines; Thyrotropin; Time; Transgenes; Transgenic Mice; Transgenic Organisms; Uterine hemorrhage; Variant; Vasomotor; Woman; Work; age related; aged; base; bone mass; design; dimer; experience; folliculogenesis; genetic analysis; glycosylation; hormone analog; improved; in vivo; inhibin; innovation; mouse model; novel; pituitary gonadal axis; poor egg quality; psychologic; reproductive; senescence; sugar

The long-term goal of this project is to study mechanisms of pituitary control of ovarian aging in women. Normal ovarian function is dependent on follicle-stimulating hormone (FSH), a pituitary derived heterodimeric glycoprotein consisting of an a and a p subunit. Both the subunits are glycosylated with two chains in each subunit. This common FSH form is called tetra-glycosylated FSH. Glycosylation plays a major role in secretion, serum half-life and biological actions of FSH. It is known that glycosylation of pituitary gonadotropins is also estrous/menstrual cycle- and age-specific. Biochemical and physiological studies have identified in several species a unique di-glycosylated variant consisting of sugar chains only in the a but not p subunits. Most importantly, the ratio of tetra- and di-glycosylated FSH is found to be age-dependant, with low levels of di-glycosylated variant predominantly present in post-menopausal women. However, the distinct in vivo biological functions of these FSH variants are unknown in normal ovarian physiology. The hypothesis is that a more potent di-glycosylated FSH is required for optimal stimulation of foliiculogenesis, but not in the aging ovary. This hypothesis will be tested in genetically engineered mouse models in three specific Aims. In Aim 1, we will test the in vivo potencies of biochemically purified FSH variants from different ages of women using FSHp knockout mice. In Aim 2, we will produce transgenic mice with a directed expression of this variant in gonadotropes and introduce this transgene into FSHp null genetic background, to test the biological actions of di-glycosylated variant. In Aim 3, we will temporally regulate the expression (gene switch) of the di-glycosylated FSH variant on a FSHp null background and identify the age-specific biological effects of di-glycosylated variant. Functional analyses with the above genetic models will identify distinct biological actions of FSH variants in vivo during ovarian aging. Ovarian follicle reserve diminishes with aging in women as well as other species and this directly affects the oocyte quality. Therefore, our studies will have a direct clinical impact in formulating new treatments for enhancing the production of healthy follicles and oocytes in aging women.

该项目的长期目标是研究女性卵巢老化的脑下垂体控制机制。 正常的卵巢功能依赖于卵泡刺激素(FSH)，这是一种来自垂体的异源二聚体 由a和p亚基组成的糖蛋白。两个亚基都是糖基化的，每个亚基上都有两条链 亚单位。这种常见的促卵泡激素形式被称为四糖基化促性腺激素。糖基化在分泌过程中起着主要作用， 促性腺激素的血清半衰期和生物学作用。已知垂体促性腺激素的糖基化也是 发情/月经周期和年龄相关。生化和生理研究已经确定了几种 一种独特的二糖基变异体，只在a亚基中含有糖链，而不在p亚基中。多数 重要的是，四糖化和二糖化卵泡刺激素的比例被发现是年龄相关的，低水平的 二糖基变异体主要存在于绝经后妇女。然而，截然不同的体内生物 这些FSH变异体的功能在正常的卵巢生理中是未知的。假设是，一个更强大的 二糖基化的卵泡刺激素是最佳的小叶发生刺激所必需的，但在老化的卵巢中不需要。这 这一假设将在基因工程小鼠模型中进行三个特定目的的测试。在目标1中，我们将测试 使用FSHp基因敲除小鼠对来自不同年龄女性的生物化学纯化的FSH变异体的体内效力。 在目标2中，我们将培育转基因小鼠，使其在促性腺激素和 将此转基因引入FSHp缺失的遗传背景，以检测二糖基化的生物学作用 变种。在目标3中，我们将在时间上调节二糖化FSH变异体在 FSHp为空背景，并鉴定二糖基变异体的年龄特异性生物学效应。 对上述遗传模型的功能分析将确定FSH变体的不同生物学行为 在体内卵巢老化的过程中。女性以及其他物种的卵巢卵泡储备随着年龄的增长而减少 这直接影响卵母细胞的质量。因此，我们的研究将对制定新的 提高老年妇女健康卵泡和卵母细胞产量的治疗方法。