Progesterone receptor regulation of seizures

孕激素受体对癫痫发作的调节

• 批准号: 10594041
• 负责人: Suchitra Joshi
• 金额: $ 35.33万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594041
• 关键词: AMPA Receptors; Acids; Age; Agonist; Allopregnanolone; Aminobutyric Acids; Animal Model; Animals; Anti-Progestin; Anticonvulsants; Biological Assay; Brain; Chemosensitization; Clinical Trials; Contraceptive Agents; Disease; Electric Stimulation; Electroencephalography; Electrophysiology (science); Enzymes; Epilepsy; Epileptogenesis; Estrous Cycle; Experimental Animal Model; Female; Fluorescent in Situ Hybridization; Frequencies; GABA-A Receptor; Generalized Epilepsy; Genetic; Genomics; Glutamates; Hippocampus; Hormonal; Incidence; Knock-out; Knockout Mice; Link; Mediating; Menstrual cycle; Mental disorders; Messenger RNA; Mifepristone; Migraine; Mus; Nestorone; Neuromodulator; Neurons; Periodicity; Pharmaceutical Preparations; Play; Predisposition; Progesterone; Progesterone Receptors; Progestins; Protein Subunits; Quantitative Reverse Transcriptase PCR; Receptor Activation; Regulation; Reporting; Research; Risk; Role; Seizures; Status Epilepticus; Synapses; Synaptic Transmission; Techniques; Testing; Up-Regulation; Western Blotting; Wild Type Mouse; Withdrawal; Woman; dysphoria; effective therapy; experience; granule cell; hippocampal pyramidal neuron; hormonal contraception; hypothalamic pituitary axis; insight; knockout animal; mRNA Expression; nervous system disorder; neuronal excitability; neurosteroids; neurotransmission; new therapeutic target; novel; perimenstrual; positive allosteric modulator; prevent; protein expression; receptor; receptor expression; reproductive; sex; text searching; transmission process

Abstract We propose to test the hypothesis that progesterone receptor (PR) activation enhances the AMPA receptor (AMPAR)-mediated synaptic transmission of hippocampal principal neurons and increases seizure susceptibility. Cyclic fluctuations in progesterone levels in women of reproductive age with epilepsy are linked to catamenial seizure exacerbation. Furthermore, perimenstrual progesterone withdrawal is linked to migraines and dysphoria. Whether PR activation regulates these disorders is not known. Progesterone is historically thought to exert anticonvulsant effects via the potentiation of GABA-A receptor- mediated inhibition. In contrast, the role of PRs, which are also expressed in the brain, in regulating neuronal activity is poorly understood outside of the hypothalamic-pituitary axis. Our preliminary studies show that PR activation increases AMPAR expression in the hippocampus and enhances the AMPAR-mediated synaptic transmission of CA1 pyramidal neurons. These changes were associated with the increased activity of CA1 neurons. Furthermore, the blockade of PR by the anti-progestin RU-486 or by the genetic deletion of PRs prevented the observed upregulation of AMPARs. Finally, the specific activation of PRs with a synthetic agonist, nestorone, increased seizure susceptibility, while RU-486 treatment suppressed neurosteroid withdrawal-induced seizures in an experimental animal model. We propose to directly test the role of PRs in regulating AMPAR expression and the excitability of hippocampal principal neurons and to assess whether PRs regulate seizure susceptibility. In the first aim, we will activate PRs using nestorone in wild-type and knockout animals and determine its effect on the neuronal excitability and AMPAR-mediated synaptic transmission of CA1 pyramidal neurons and dentate granule cells, and on the expression of GluA1 and GluA2 subunit proteins and mRNA. We will use western blotting, qRT-PCR, and electrophysiological techniques in these studies. In the second aim, we will determine whether PR expression in CA1 neurons and DGCs changes during the estrous cycle, using fluorescent in situ hybridization assay, qRT-PCR and a mouse expressing myc-tagged PR. In the third aim, we will test whether PR activation reduces the seizure threshold and alters the susceptibility to status epilepticus and epileptogenesis by using continuous video-EEG recording.

抽象的 我们建议测试孕酮受体（PR）激活增强AMPA受体的假设 （AMPAR）介导的海马主神经元的突触传播，并增加癫痫发作 敏感性。癫痫生殖年龄妇女的孕激素水平的环状波动与癫痫 为癫痫发作加剧。此外，妊娠孕酮的戒断与偏头痛有关 和烦躁不安。 PR激活是否可以调节这些疾病。 从历史上看，孕酮通过增强GABA-A受体 - 介导的抑制作用。相反，在调节神经元中，PR的作用也在大脑中表达 在下丘脑 - 垂体轴之外，活动知之甚少。我们的初步研究表明PR 激活增加海马中的AMPAR表达并增强AMPAR介导的突触 CA1锥体神经元的传播。这些变化与CA1的活性增加有关 神经元。此外，抗progestin ru-486或PR的遗传缺失对PR的封锁 阻止了观察到的AMPAR上调。最后，具有合成的PR的特异性激活 激动剂，Nestorone，提高了癫痫敏感性，而RU-486治疗抑制了神经类似 在实验动物模型中提取引起的癫痫发作。 我们建议直接测试PR在调节AMPAR表达和兴奋性中的作用 海马主要神经元，并评估PR是否调节癫痫发作易感性。 在第一个目标中，我们将使用野生型和淘汰动物的Nestorone激活PR，并确定其 对CA1锥体神经元的神经元兴奋性和AMPAR介导的突触传播的影响 齿状颗粒细胞，以及GluA1和GluA2亚基蛋白和mRNA的表达。我们将使用 这些研究中的蛋白质印迹，QRT-PCR和电​​生理技术。在第二个目标中，我们将 使用使用 荧光原位杂交测定法，QRT-PCR和表达MYC标记的PR的小鼠。在第三个目标中，我们 将测试PR激活是否会降低癫痫发作阈值并改变对癫痫状态的敏感性 通过使用连续的视频录制来进行癫痫发生。