The Aging Pituitary/Gonadal Axis

衰老的垂体/性腺轴

基本信息

批准号：
10627088
负责人：
GEORGE R BOUSFIELD
金额：
$ 226.37万
依托单位：
WICHITA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-15 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10627088
关键词：
ATAC-seq Address Affect Age Aging Agonist Alternative Splicing American Antral Arrestins Back Binding Binding Sites Biochemical Biological Blood Bone Density CREB1 gene Cell Line Cell membrane Cells Circulation Confusion Coupled Coupling Cryoelectron Microscopy Cyclic AMP Development Disparate Elderly Engineering Enzyme-Linked Immunosorbent Assay Enzymes Estrogens Evaluation Exhibits Feedback Female Fertility Fertility Agents Follicle Stimulating Hormone Follicle Stimulating Hormone Receptor Funding GTP-Binding Proteins Genes Genetic Models Genetic Transcription Genitourinary system Glycoproteins Goals Green Fluorescent Proteins Health Hormone Receptor Hormones Human Impairment Infertility Injections Knock-out Knockout Mice Knowledge LH Receptors Laboratories Lifting Ligand Binding Link Lipids Location Measures Mediating Membrane Membrane Proteins Menstrual cycle Molecular Molecular Conformation Morbidity - disease rate Mus N-Glycosylation Site Obesity Oligosaccharides Organism Osteoclasts Osteoporosis Outcome Outcomes Research Ovarian Ovarian Granulosa Cell Ovarian aging Ovary Pathway interactions Pattern Perimenopause Physiological Pituitary Gland Play Polysaccharides Preparation Process Progesterone Protein Hormone Receptor Reagent Receptor Activation Reproduction Research Role Rotation Side Signal Pathway Signal Transduction Steroid Receptors Steroids Structure System Thyrotropin Receptor Tissues Transgenic Mice Transgenic Organisms Uterine hemorrhage Variant Whole Organism Woman age effect age related aged bone bone loss bone mass female reproductive system fertility improvement glycosylation granulosa cell human female hypothalamic pituitary ovarian axis improved in vivo infertility treatment inhibin monomer mouse genetics mouse model mullerian-inhibiting hormone nanodisk older women programs psychologic receptor reproductive reproductive senescence response single-cell RNA sequencing structural biology trafficking transcription factor transcriptome sequencing translational potential vasomotor symptoms young woman

项目摘要

Project Summary/Abstract - Overall This integrated program project was pursuant to the discovery of naturally occurring, partially glycosylated follicle-stimulating hormone (FSH) glycoforms by the Bousfield laboratory. Two of these glycoforms possessed 3 of 4 N-glycans, exhibited greater biological activity, and were more abundant than fully-glycosylated FSH in young women. Furthermore, hypo-glycosylated FSH variants exhibited an age-dependent decrease concomitant with a well-known decline in fertility in women. Projects are organized to reflect the recognition of the importance of the development of numerous mouse genetic models in previous funding periods inspired by biochemical advances in understanding of the FSH glycoforms, and which now, will inspire in turn, such studies at the signaling, trafficking, and structural biology levels. In Project 1(Kumar), powerful genetic models now make it possible to study each specific glycoform independently, both by injection of purified glycoform preparations into Fshb-null mice as well as in transgenic mice expressing FSH18, FSH21, or FSH24. Advances in single-cell RNA-sequencing coupled with a transgenic line that expresses green fluorescent protein specifically in gonadotropes permit evaluation of the more than 50 enzymes responsible for N-linked oligosaccharide synthesis. Since non-ovarian targets of FSH have raised concerns and confusion for women undergoing IVF, several mouse models were devised to address questions raised by conflicting experimental outcomes, beginning with deletion of the Fshr gene, which would effectively eliminate all forms of putative FSHR proteins, including alternative splicing. In Project 2(Davis), studies involving FSH glycoform signals within primary cultures of granulosa cells, focus on the disparate responses of ovarian target cells to biased agonists FSH18, FSH21, and FSH24. Two major, differentially glycoform-regulated transcription factors (TF), CREB and YAP1, will have their downstream pathways characterized in granulosa cells. Granulosa cells of young and advanced age mice and women will provide basic information relevant to translational efforts. The interface between FSH and FSHR, both monomeric and oligomeric receptor forms, will be studied in Project 3(Jonas). Activation of cAMP accumulation is now known to involve FSHR dynamics that will be evaluated, including internalization. FSH glycoforms are anticipated to alter FSHR conformation to varying degrees, leading to biased agonist signals. Project 4(Bousfield) will incorporate FSHR into lipid nanodiscs to provide a means to evaluate glycoform-FSHR- membrane interactions, using cryogenic electron microscopy (cyro-EM). This has been done with LH and TSH receptors (LHR & TSHR) and those studies have revealed the ligand binding site undergoes a major rotation, lifting it away from the membrane, during receptor activation. The location of FSH oligosaccha-rides is on the back side of the hormone, away from the hormone-receptor interface, which suggests interaction with the cell membrane or membrane proteins. Taken together, the projects integrate FSH glycoform signaling at overlapping levels ranging from the intact organism to the glycosylated hormone itself.

项目摘要/摘要 - 总体这个集成的程序项目是根据发现自然发生的部分糖基化的 Bousfield实验室刺激卵泡激素（FSH）糖型。这些糖衣中的两个拥有3 在4种N-聚糖中表现出更大的生物学活性，并且比年轻的糖基化FSH更丰富女性。此外，低糖基化的FSH变体表现出与年龄相关的降低。女性生育能力的众所周知。组织的项目是为了反映对重要性的认识在以前的资金期间的众多遗传模型的发展受生物化学启发理解FSH糖型的进步，现在将激发这种研究信号传导，贩运和结构生物学水平。在项目1（库马尔）中，有力的遗传模型现在成为可以独立研究每个特定的糖型 FSHB无效的小鼠以及表达FSH18，FSH21或FSH24的转基因小鼠。单细胞的进步 RNA测序与转基因线结合，该系在促性腺激素允许评估50多种负责N连接寡糖的酶合成。由于FSH的非肥大目标引起了人们对参加IVF的妇女的关注和困惑，因此设计了几种鼠标模型来解决通过冲突的实验结果提出的问题，从删除FSHR基因开始，该基因将有效地消除所有形式的假定FSHR蛋白，包括替代剪接。在项目2（戴维斯）中，涉及初级文化中FSH糖型信号的研究颗粒细胞的侧重于卵巢靶细胞对偏见激动剂FSH18，FSH21和 FSH24。两个主要的，差异性糖剂调节的转录因子（TF），CREB和YAP1将具有其在颗粒细胞中表征的下游途径。年轻小鼠和高龄小鼠的颗粒细胞以及妇女将提供与翻译工作有关的基本信息。 FSH和FSHR之间的接口将在项目3（Jonas）中研究单体和低聚受体形式。营地积累的激活现在已知涉及将评估的FSHR动力学，包括内部化。 FSH糖型是预计会在不同程度上改变FSHR构象，从而导致偏见的激动剂信号。项目 4（Bousfield）将将FSHR纳入脂质纳米盘中，以提供一种评估糖型-fshr-的方法膜相互作用，使用低温电子显微镜（Cyro-EM）。这是通过LH和TSH完成的受体（LHR和TSHR）和这些研究表明，配体结合位点发生了重大旋转，在受体激活期间，将其从膜上提起。 FSH寡果骑兵的位置在激素的背面，远离激素受体界面，这表明与细胞相互作用膜或膜蛋白。综上从完整生物到糖基化激素本身的重叠水平。

项目成果

期刊论文数量（60）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Molecular regulation of follicle-stimulating hormone synthesis, secretion and action.

DOI：
10.1530/jme-17-0308
发表时间：
2018-04
期刊：
Journal of molecular endocrinology
影响因子：
3.5
作者：
Das N;Kumar TR
通讯作者：
Kumar TR

The SO(H)L(H) "O" drivers of oocyte growth and survival but not meiosis I.

SO(H)L(H)“O”驱动卵母细胞生长和存活，但不是减数分裂 I。

DOI：
10.1172/jci94665
发表时间：
2017
期刊：
The Journal of clinical investigation
影响因子：
0
作者：
Kumar,TRajendra
通讯作者：
Kumar,TRajendra

Partially deglycosylated equine LH preferentially activates beta-arrestin-dependent signaling at the follicle-stimulating hormone receptor.

部分去糖基化的马 LH 优先激活促卵泡激素受体上的 β-抑制蛋白依赖性信号传导。

DOI：
10.1210/me.2009-0347
发表时间：
2010
期刊：
Molecular endocrinology (Baltimore, Md.)
影响因子：
0
作者：
Wehbi,Vanessa;Tranchant,Thibaud;Durand,Guillaume;Musnier,Astrid;Decourtye,Jérémy;Piketty,Vincent;Butnev,VladimirY;Bousfield,GeorgeR;Crépieux,Pascale;Maurel,Marie-Christine;Reiter,Eric
通讯作者：
Reiter,Eric

Prostaglandin F2α regulates mitochondrial dynamics and mitophagy in the bovine corpus luteum.