Intercellular dynamics of cyclic nucleotides in ovarian follicles
卵巢卵泡中环核苷酸的细胞间动力学
基本信息
- 批准号:10380740
- 负责人:
- 金额:$ 8.2万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-04-01 至 2023-09-30
- 项目状态:已结题
- 来源:
- 关键词:AddressAdenylate CyclaseAttenuatedBasal laminaBindingBreedingCarbenoxoloneCellsClinicalCommunicationConfocal MicroscopyCrossbreedingCyclic AMPCyclic AMP-Dependent Protein KinasesCyclic GMPCyclic NucleotidesDiffuseDiffusionDoseEnvironmentEpidermal Growth Factor ReceptorEpidermal Growth Factor Receptor Tyrosine Kinase InhibitorEpitopesExhibitsExposure toFamilyFluorescence Resonance Energy TransferFutureGap JunctionsHormone useHourHumanImageIn VitroIncubatedIndividualKnowledgeLH ReceptorsLeadLightLuteinizationLuteinizing HormoneMammalsMeasurementMediatingMeiosisMicroscopeMicroscopyMusOocytesOutcomeOvarian FollicleOvaryOvulationPKA inhibitorPatternPhosphorylationPhysiologicalPhysiological ProcessesProductionReceptor ActivationReproductive HealthResearchResolutionSecond Messenger SystemsSignal TransductionSomatic CellSuggestionTestingTimeTissuesToxic effectegggranulosa cellhormonal signalsinfertility treatmentinhibitornovel strategiesoocyte maturationpreventresponsesensorspatiotemporal
项目摘要
Project Summary/Abstract
In mammalian ovaries, the mid-cycle surge of luteinizing hormone (LH) acts on the granulosa cells of
preovulatory follicles to trigger oocyte maturation, ovulation, and luteinization. Signaling through LH receptors
occurs primarily occurs through sequential Gs-mediated adenylyl cyclase activation, production of the second
messenger cAMP, and protein kinase A-dependent phosphorylation. However, whether and how the cAMP
signal spreads from the subset of cells that express the LH receptor (LHR) to achieve the many outcomes of
the LH surge is a long-standing question that remains to be resolved. To test this and other questions, intact
preovulatory follicles will be isolated from mice that globally express a newly developed fluorescent sensor for
cAMP called R-FlincA that can increase brightness by up to 600% upon cAMP binding. Using R-FlincA and
state-of-the-art light sheet microscopy, individual cells can be imaged as they elevate cAMP either directly in
response to LH, or by diffusion between cells. If diffusion is detected, it is hypothesized to be via gap junctions,
which connect all granulosa cells to each other, and to the oocyte. To test this, follicles will be incubated in a
gap junction inhibitor, carbenoxolone, prior to LH treatment. It is hypothesized that only some cells will elevate
cAMP, corresponding to LHR-expressing cells, with no diffusion between cells. Diffusion of cAMP through gap
junctions would present a paradox because the LH surge also causes a decrease in cAMP in the oocyte, which
is necessary for meiotic resumption. It is hypothesized that LH signaling through epidermal growth factor
receptor (EGFR) kinase closes gap juctions between granulosa cells and the oocyte, allowing cAMP to decline
in the oocyte while remaining elevated in granulosa cells. This will be tested using AG1478, an inhibitor of
EGFR kinase activity. In the presence of AG1478, it is predicted that cAMP will continue to diffuse through gap
junctions from granulosa cells into the oocyte, which would prevent oocyte maturation. The LH surge also
causes the rapid decrease of another cyclic nucleotide, cGMP, in the granulosa cells and oocyte. This
decrease in oocyte cGMP allows for the decrease in oocyte cAMP, which then triggers oocyte maturation into a
fertilizable egg. However, it is not known how the granulosa cell cAMP increase and cGMP decrease are
related. This will be investigated by breeding R-FlincA mice with another mouse line that expresses a
fluorescent sensor for cGMP. It is hypothesized that cells that initially elevate cAMP in response to LH will
rapidly exhibit decreased cGMP levels, suggesting that the cAMP increase is required to lower cGMP levels.
This project will contribute to understanding of how LH signaling leads to a fertilizable egg, and could lead to
improvements in human in vitro maturation (IVM), which depends on optimal cyclic nucleotide levels in the
oocyte. Thus this project could lead to clinical advances in reproductive health.
项目概要/摘要
在哺乳动物卵巢中,黄体生成素 (LH) 的周期中期激增作用于颗粒细胞
排卵前卵泡触发卵母细胞成熟、排卵和黄体化。通过 LH 受体发出信号
主要通过连续 Gs 介导的腺苷酸环化酶激活发生,产生第二个
信使 cAMP 和蛋白激酶 A 依赖性磷酸化。然而,cAMP 是否以及如何
信号从表达 LH 受体 (LHR) 的细胞子集传播,以实现许多结果
LH 激增是一个长期存在的问题,有待解决。为了测试这个和其他问题,完好无损
将从小鼠体内分离出排卵前卵泡,这些卵泡在整体上表达新开发的荧光传感器
cAMP 称为 R-FlincA,在 cAMP 结合后可将亮度提高高达 600%。使用 R-FlincA 和
最先进的光片显微镜,可以在单个细胞升高 cAMP 时直接对它们进行成像
对 LH 的反应,或通过细胞之间的扩散。如果检测到扩散,则假设是通过间隙连接,
它将所有颗粒细胞相互连接并与卵母细胞连接。为了测试这一点,将卵泡培养在
LH 治疗前,间隙连接抑制剂,carbenoxolone。据推测,只有部分细胞会升高
cAMP,对应于 LHR 表达细胞,细胞间无扩散。 cAMP 通过间隙扩散
连接处会出现一个悖论,因为 LH 激增也会导致卵母细胞中 cAMP 的减少,从而
对于减数分裂恢复是必要的。据推测,LH 信号通过表皮生长因子
受体 (EGFR) 激酶关闭颗粒细胞和卵母细胞之间的间隙连接,使 cAMP 下降
在卵母细胞中,而在颗粒细胞中保持升高。这将使用 AG1478 进行测试,AG1478 是一种抑制剂
EGFR 激酶活性。在 AG1478 存在的情况下,预计 cAMP 将继续通过间隙扩散
颗粒细胞与卵母细胞的连接,这会阻止卵母细胞成熟。 LH 也激增
导致颗粒细胞和卵母细胞中另一种环核苷酸 cGMP 的快速减少。这
卵母细胞 cGMP 的减少导致卵母细胞 cAMP 的减少,从而触发卵母细胞成熟为
受精卵。然而,尚不清楚颗粒细胞cAMP增加和cGMP减少是如何发生的。
有关的。这将通过将 R-FlincA 小鼠与表达
cGMP 荧光传感器。据推测,最初响应 LH 升高 cAMP 的细胞将
迅速表现出 cGMP 水平下降,表明需要增加 cAMP 来降低 cGMP 水平。
该项目将有助于了解 LH 信号传导如何导致受精卵,并可能导致
人类体外成熟(IVM)的改善,这取决于体内的最佳环核苷酸水平
卵母细胞。因此,该项目可能会导致生殖健康方面的临床进展。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Jeremy R. Egbert其他文献
Jeremy R. Egbert的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Jeremy R. Egbert', 18)}}的其他基金
Intercellular dynamics of cyclic nucleotides in ovarian follicles
卵巢卵泡中环核苷酸的细胞间动力学
- 批准号:
10195538 - 财政年份:2021
- 资助金额:
$ 8.2万 - 项目类别:
相似海外基金
Neuroendocrine regulation of energy metabolism: role of pituitary adenylate cyclase-activating polypeptide (PACAP) in the thermoregulatory cascade
能量代谢的神经内分泌调节:垂体腺苷酸环化酶激活多肽(PACAP)在温度调节级联中的作用
- 批准号:
RGPIN-2021-04040 - 财政年份:2022
- 资助金额:
$ 8.2万 - 项目类别:
Discovery Grants Program - Individual
Controlled Release of Pituitary Adenylate Cyclase Activating Polypeptide from a Hydrogel-Nanoparticle Delivery Vehicle for Applications in the Central Nervous System
从水凝胶-纳米粒子递送载体中控制释放垂体腺苷酸环化酶激活多肽,用于中枢神经系统的应用
- 批准号:
547124-2020 - 财政年份:2022
- 资助金额:
$ 8.2万 - 项目类别:
Alexander Graham Bell Canada Graduate Scholarships - Doctoral
Controlled Release of Pituitary Adenylate Cyclase Activating Polypeptide from a Hydrogel-Nanoparticle Delivery Vehicle for Applications in the Central Nervous System
从水凝胶-纳米粒子递送载体中控制释放垂体腺苷酸环化酶激活多肽,用于中枢神经系统的应用
- 批准号:
547124-2020 - 财政年份:2021
- 资助金额:
$ 8.2万 - 项目类别:
Postgraduate Scholarships - Doctoral
Neuroendocrine regulation of energy metabolism: role of pituitary adenylate cyclase-activating polypeptide (PACAP) in the thermoregulatory cascade
能量代谢的神经内分泌调节:垂体腺苷酸环化酶激活多肽(PACAP)在温度调节级联中的作用
- 批准号:
RGPIN-2021-04040 - 财政年份:2021
- 资助金额:
$ 8.2万 - 项目类别:
Discovery Grants Program - Individual
The Molecular Mechanism of the Secretion of the Bacterial Toxin Adenylate Cyclase
细菌毒素腺苷酸环化酶分泌的分子机制
- 批准号:
451966 - 财政年份:2021
- 资助金额:
$ 8.2万 - 项目类别:
Operating Grants
The role of prefrontostriatal Pituitary Adenylate Cyclase Activating Polypeptide in excessive and compulsive ethanol drinking
前额纹状体垂体腺苷酸环化酶激活多肽在过量和强迫性乙醇饮酒中的作用
- 批准号:
10455587 - 财政年份:2020
- 资助金额:
$ 8.2万 - 项目类别:
The role of prefrontostriatal Pituitary Adenylate Cyclase Activating Polypeptide in excessive and compulsive ethanol drinking
前额纹状体垂体腺苷酸环化酶激活多肽在过量和强迫性乙醇饮酒中的作用
- 批准号:
10261394 - 财政年份:2020
- 资助金额:
$ 8.2万 - 项目类别:
Diagnosis and therapeutic effect of neurally mediated syncope (NMS) using fluctuation of adenylate cyclase activity
利用腺苷酸环化酶活性波动对神经介导性晕厥(NMS)的诊断和治疗效果
- 批准号:
20K08498 - 财政年份:2020
- 资助金额:
$ 8.2万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Pituitary adenylate cyclase-activating polypeptide 27 in the paraventricular thalamus and its projections: Role in ethanol drinking
室旁丘脑中的垂体腺苷酸环化酶激活多肽 27 及其预测:在乙醇饮用中的作用
- 批准号:
10380126 - 财政年份:2020
- 资助金额:
$ 8.2万 - 项目类别:
The role of prefrontostriatal Pituitary Adenylate Cyclase Activating Polypeptide in excessive and compulsive ethanol drinking
前额纹状体垂体腺苷酸环化酶激活多肽在过量和强迫性乙醇饮酒中的作用
- 批准号:
10662279 - 财政年份:2020
- 资助金额:
$ 8.2万 - 项目类别: