Progesterone receptor regulation of seizures

孕激素受体对癫痫发作的调节

基本信息

批准号：
9886300
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
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9886300
关键词：
AMPA Receptors Acids Age Agonist Allopregnanolone Aminobutyric Acids Animal Model Animals Anti-Progestin Anticonvulsants Biological Assay Brain Chemosensitization Clinical Trials 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 (Brain)Hormonal Incidence Knock-out Knockout Mice Lead Link Literature 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 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 transmission process

项目摘要

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.

抽象的我们建议检验黄体酮受体 (PR) 激活增强 AMPA 受体的假设 (AMPAR) 介导的海马主要神经元突触传递并增加癫痫发作易感性。育龄妇女孕酮水平的周期性波动与癫痫有关导致经期癫痫发作加剧。此外，经期黄体酮戒断与偏头痛有关和烦躁不安。 PR 激活是否调节这些疾病尚不清楚。历史上黄体酮被认为通过增强 GABA-A 受体发挥抗惊厥作用介导的抑制。相比之下，也在大脑中表达的 PR 在调节神经元中的作用人们对下丘脑-垂体轴之外的活动知之甚少。我们的初步研究表明，公关激活增加海马 AMPAR 表达并增强 AMPAR 介导的突触 CA1 锥体神经元的传递。这些变化与 CA1 活性增加有关神经元。此外，抗孕激素 RU-486 或 PR 基因删除可阻断 PR 阻止了观察到的 AMPAR 上调。最后，用合成的 PR 进行特异性激活激动剂nestorone，增加癫痫易感性，而RU-486治疗抑制神经类固醇实验动物模型中戒断诱发的癫痫发作。我们建议直接测试 PR 在调节 AMPAR 表达和 AMPAR 兴奋性中的作用海马主要神经元并评估 PR 是否调节癫痫易感性。第一个目标是，我们将在野生型和基因敲除动物中使用nestorone激活PR，并确定其对神经元兴奋性和 AMPAR 介导的 CA1 锥体神经元突触传递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 和电生理学技术。在第二个目标中，我们将确定 CA1 神经元和 DGC 中的 PR 表达在动情周期期间是否发生变化，使用荧光原位杂交测定、qRT-PCR 和表达 myc 标记 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阈值并改变对癫痫持续状态的易感性通过使用连续视频脑电图记录和癫痫发生。