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: 8377843
• 负责人: T. RAJENDRA KUMAR
• 金额: $ 21.73万
• 依托单位: WICHITA STATE UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8377843
• 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; analog; base; bone mass; design; dimer; experience; folliculogenesis; genetic analysis; glycosylation; improved; in vivo; inhibin; innovation; mouse model; novel; pituitary gonadal axis; poor egg quality; psychologic; reproductive; senescence; sugar; young woman

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衰老的垂体控制机制。 正常的卵巢功能取决于刺激激素（FSH），这是一种垂体衍生的异二聚体 由A和P亚基组成的糖蛋白。两个亚基都用两个链中的两个链糖基化 亚基。这种常见的FSH形式称为四糖基化FSH。糖基化在分泌中起主要作用， 血清半衰期和FSH的生物作用。众所周知，垂体促性腺激素的糖基化也是 发情/月经周期和年龄特异性。生化和生理研究已经确定 物种一种独特的Di-糖基化变体，仅在A中但不是P亚基中的糖链组成。最多 重要的是，发现四 - 二糖基化的FSH的比率是年龄依赖的，低水平 二糖基化变体主要存在于绝经后妇女中。然而，独特的体内生物学 这些FSH变体的功能在正常的卵巢生理学中是未知的。假设是一个更有效的 二糖基化的FSH是最佳刺激叶状发生所必需的，但在卵巢老龄化中不需要。这 假设将以三个特定目的的基因工程小鼠模型进行检验。在AIM 1中，我们将测试 使用FSHP基因敲除小鼠，来自不同年龄的女性的生化纯化的FSH变体的体内效力。 在AIM 2中，我们将在促性腺激素中以这种变体的定向表达产生转基因小鼠和 将此转基因引入FSHP无效的遗传背景，以测试二糖基化的生物学作用 变体。在AIM 3中，我们将暂时调节A上的Di-糖基化FSH变体的表达（基因开关） FSHP无效背景并确定二糖基化变体的年龄特异性生物学作用。 使用上述遗传模型的功能分析将确定FSH变体的不同生物学作用 卵巢老化期间的体内。卵巢卵泡储备会减少妇女以及其他物种的衰老，以及 这直接影响卵母细胞质量。因此，我们的研究将在制定新的方面具有直接的临床影响 增强衰老女性健康卵泡和卵母细胞的生产的治疗方法。