Age-Dependent N-Glycosylation of Follicle-Stimulation Hormone in Gonadotropes

促性腺激素中卵泡刺激激素的年龄依赖性 N-糖基化

• 批准号: 10679254
• 负责人: Rosemary McDonald
• 金额: $ 1.97万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10679254
• 关键词: Address; Adipose tissue; Affect; Age; Aging; Anterior Pituitary Gland; Binding; Biochemical; Bioinformatics; Biological; Biological Assay; Bone Tissue; Cardiovascular Diseases; Cells; Clinical Treatment; Complex; Cre driver; Data; ESR1 gene; Enzymes; Estrogen Receptors; Estrogen decline; Estrogens; Etiology; Event; Exhibits; Feedback; Female; Fluorescence-Activated Cell Sorting; Follicle Stimulating Hormone; Gene Expression; Genes; Genetic; Glycoproteins; Goals; Gonadotrope Cell; Hormones; Human; Immunofluorescence Immunologic; Inflammatory; Investigation; Knock-out; Knowledge; Life Style; Longevity; LoxP-flanked allele; Luteinizing Hormone; Menopausal Symptom; Menopause; Messenger RNA; Metabolic Diseases; Molecular; Mus; Obesity; Organism; Osteoporosis; Ovarian; Ovarian Steroid Hormone; Ovarian aging; Pathway interactions; Perimenopause; Phenotype; Physiological; Pituitary Gland; Postmenopause; Premenopause; Process; Progesterone; Progesterone Receptors; Proteins; Receptor Signaling; Regulation; Research; Role; Signal Transduction; Stains; Steroid Receptors; Steroids; Symptoms; Tissues; Up-Regulation; Urine; Visceral; Weight Gain; Woman; Work; age related; aged; body system; bone; bone loss; common symptom; enzyme activity; enzyme pathway; experience; glycosylation; human old age (65+); hypothalamic pituitary gonadal axis; in vivo; mRNA Expression; middle age; mouse model; novel; older women; promoter; protein expression; receptor-mediated signaling; reproductive; reproductive senescence; steroid hormone receptor; targeted treatment; transcriptome sequencing; young woman; Address; Adipose tissue; Affect; Age; Aging; Anterior Pituitary Gland; Binding; Biochemical; Bioinformatics; Biological; Biological Assay; Bone Tissue; Cardiovascular Diseases; Cells; Clinical Treatment; Complex; Cre driver; Data; ESR1 gene; Enzymes; Estrogen Receptors; Estrogen decline; Estrogens; Etiology; Event; Exhibits; Feedback; Female; Fluorescence-Activated Cell Sorting; Follicle Stimulating Hormone; Gene Expression; Genes; Genetic; Glycoproteins; Goals; Gonadotrope Cell; Hormones; Human; Immunofluorescence Immunologic; Inflammatory; Investigation; Knock-out; Knowledge; Life Style; Longevity; LoxP-flanked allele; Luteinizing Hormone; Menopausal Symptom; Menopause; Messenger RNA; Metabolic Diseases; Molecular; Mus; Obesity; Organism; Osteoporosis; Ovarian; Ovarian Steroid Hormone; Ovarian aging; Pathway interactions; Perimenopause; Phenotype; Physiological; Pituitary Gland; Postmenopause; Premenopause; Process; Progesterone; Progesterone Receptors; Proteins; Receptor Signaling; Regulation; Research; Role; Signal Transduction; Stains; Steroid Receptors; Steroids; Symptoms; Tissues; Up-Regulation; Urine; Visceral; Weight Gain; Woman; Work; age related; aged; body system; bone; bone loss; common symptom; enzyme activity; enzyme pathway; experience; glycosylation; human old age (65+); hypothalamic pituitary gonadal axis; in vivo; mRNA Expression; middle age; mouse model; novel; older women; promoter; protein expression; receptor-mediated signaling; reproductive; reproductive senescence; steroid hormone receptor; targeted treatment; transcriptome sequencing; young woman

PROJECT SUMMARY The menopausal transition represents a complex and dynamic physiological aging phenomenon and often leads to adverse life-style effects. Nearly 85% of post-menopausal women will experience multiple symptoms. Treatment of clinical symptoms in post-menopausal women is difficult without knowing the underlying biological mechanisms and molecular players and hence requires further investigation. The anterior pituitary-derived heterodimeric glycoprotein hormone, follicle-stimulating hormone (FSH) consists of an alpha and a beta subunit. The FSH subunit undergoes significant age-dependent changes in N-glycosylation. Higher ratios of fully glycosylated FSHβ subunit in older, post-menopausal women compared to younger, reproductively active women were identified in human pituitaries and urine. FSH levels rise significantly due to concomitantly declining ovarian estrogen and progesterone during menopause and high levels of old age specific FSH may be deleterious in tissues such as bone and adipose, where FSH acts noncanonically via inflammatory pathways. This observation is of great significance because osteoporosis and weight gain are two of the most common symptoms post-menopausal women experience. This project focuses on the N-glycosylation process within anterior pituitary gonadotrope cells and seeks to identify the molecular players involved in the “glycosylation shift” to a more fully glycosylated phenotype in older age females, as well as the factors that regulate this process. We have recently identified differentially regulated N-glycosylation enzymes expressed in both young (~4- months) and older age (8-month) female mouse gonadotrope cells via RNA-sequencing analysis. Several glycosylation pathway-encoding genes (for example, Man2a1, B4galt5) exhibited a significant upregulation in gonadotropes of older age mice. We hypothesize that these N-glycosylation enzyme- encoding genes may contribute to the age-related “glycosylation shift” in FSHβ seen in older, post-reproductively active female mice. In Specific Aim 1, we will define age-dependent changes in expression of N-glycosylation enzymes MAN2A1 and B4GALT4 in gonadotrope cells of young and old female mice (using a GFP-tagged gonadotrope mouse model) and immunostaining whole pituitary and gonadotrope cells, as well as performing enzyme activity assays. In Specific Aim 2, we will determine the role of estrogen and progesterone receptor signaling in regulation of the N- glycosylation pathway-encoding enzymes in gonadotrope cells of young and old female mice using gonadotrope-specific knockout of either Esr1 or Pgr and performing fluorescence-activated cell sorting (FACS), RNA-sequencing, qPCR, immunostaining, and enzyme activity assays in pituitary gonadotrope cells. Ultimately, the work proposed here represents a novel genetic and biochemical approach and provide new knowledge on the regulation of FSHβ N-glycosylation during reproductive aging and how this may indirectly contribute to the etiology of the well-known menopausal symptoms such as bone loss and adiposity.

项目摘要 更年期过渡代表了一种复杂而动态的物理老化现象，并且经常引导 不利的生活方式。近85％的绝经后妇女会出现多种症状。 绝经后妇女的临床症状治疗很难不知道潜在的生物学 机制和分子参与者，因此需要进一步研究。前垂体衍生的 异二聚体糖蛋白激素，诱导叶质激素（FSH）由α和β亚基组成。 FSH亚基在N-糖基化中经历了显着依赖年龄的变化。较高的比率 与年轻的，生殖活性相比 妇女在人类的垂体和尿液中被发现。 FSH水平由于同时下降而显着上升 绝经期和高年龄特异性FSH期间的卵巢雌激素和孕酮可能是 在骨骼和脂肪等组织中有害，其中FSH通过炎症途径非规范起作用。 该观察结果非常重要，因为骨质疏松和体重增加是最常见的两个 绝经后妇女经历症状。该项目侧重于内部的N-糖基化过程 垂体前促性腺纤维细胞，并试图识别涉及“糖基化的分子参与者 转移到老年女性中更完全糖基化的表型，以及调节该过程的因素。 我们最近确定了在两个年轻人中表达的差异调节的N-糖基化酶（〜4-- 几个月）和年龄（8个月）通过RNA测序分析，雌性小鼠促性腺激素细胞。一些 糖基化途径编码基因（例如，MAN2A1，B4GALT5）暴露了明显的上调 老年小鼠的促性腺激素。我们假设这些N-糖基化酶编码基因可能 在年龄较大的雌性活性雌性小鼠中，有助于年龄相关的“糖基化转移”。 在特定目标1中，我们将定义N-糖基化酶MAN2A1表达的年龄依赖性变化 和年轻雌性小鼠的促性腺细胞中的B4GALT4（使用GFP标记的性腺小鼠 模型）和免疫染色整个垂体和性腺细胞，以及进行酶活性测定。 在特定的目标2中，我们将确定雌激素和孕激素受体信号在调节中的作用 使用N-糖基化途径编码酶的年轻雌性小鼠的促性腺细胞中的酶 ESR1或PGR的性腺特异性敲除，并执行荧光激活的细胞排序（FACS）， RNA测序，QPCR，免疫染色和酶活性测定在垂体性促性腺细胞中。最终， 这里提出的工作代表了一种新颖的遗传和生化方法，并提供了有关的新知识 在生殖衰老期间的FSHβN-糖基化调节，这可能间接有助于 众所周知的更年期症状的病因，例如骨质流失和肥胖。