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垂体控制女性卵巢衰老的机制。 正常的卵巢功能依赖于促卵泡激素（FSH），一种垂体衍生的异二聚体 由a和p亚基组成的糖蛋白。两个亚基均被糖基化，每个亚基有两条链 亚基。这种常见的 FSH 形式称为四糖基化 FSH。糖基化在分泌中起重要作用， FSH 的血清半衰期和生物学作用。据了解，垂体促性腺激素的糖基化也 发情/月经周期和年龄特定。生化和生理学研究已经确定了一些 物种是一种独特的二糖基化变体，仅在 a 亚基中包含糖链，但在 p 亚基中不包含糖链。最多 重要的是，四糖基化和二糖基化 FSH 的比例被发现与年龄有关，且 FSH 水平较低。 二糖基化变体主要存在于绝经后妇女中。然而，不同的体内生物 这些 FSH 变体的功能在正常卵巢生理学中是未知的。假设有一个更有效的 二糖基化 FSH 是最佳刺激卵泡生成所必需的，但在衰老的卵巢中则不需要。这 该假设将在三个特定目标的基因工程小鼠模型中进行测试。在目标 1 中，我们将测试 使用 FSHp 敲除小鼠对来自不同年龄女性的生化纯化 FSH 变体进行体内效力。 在目标 2 中，我们将生产在促性腺激素和促性腺激素中定向表达该变体的转基因小鼠 将此转基因引入FSHp无效遗传背景，以测试二糖基化的生物学作用 变体。在目标 3 中，我们将暂时调节二糖基化 FSH 变体在 FSHp 零背景并鉴定二糖基化变体的年龄特异性生物学效应。 使用上述遗传模型进行功能分析将识别 FSH 变体的独特生物学作用 卵巢老化期间的体内。女性和其他物种的卵巢卵泡储备会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药物产生直接的临床影响。 增强老年女性健康卵泡和卵母细胞生成的治疗方法。