Epilepsy-Associated Dysfunction in the Kisspeptin-GnRH Neural Circuit

项目总结 颞叶癫痫(TLE)是成人最常见的局灶性癫痫，患者有 生殖内分泌紊乱的发生率高于一般人群。值得注意的是， 临床研究还发现，无论男女，黄体生成素(LH)的释放模式都受到了干扰。 促性腺激素释放激素(GnRH)神经元形成大脑控制 生殖，它们在脑下垂体释放黄体生成素的过程中起关键作用。我们的实验室表明，促性腺激素释放激素神经元 海马海人酸(IHKA)模型小鼠的放电活动和内源性兴奋性被破坏 TLE。弓状核(ARC)和脑室周围前腹侧的Kispeptin上游神经元亚群 AVPV是GnRH神经元突触传递的主要来源。弧形kispeptin神经元扮演着一个 黄体生成素在雌雄交界性黄体生成素的脉动性释放中起关键作用，而动静脉曲张蛋白神经元调节排卵前 促性腺激素释放激素/促黄体生成素在女性中激增。然而，Kispeptin神经元功能从来没有在动物模型中被研究过。 癫痫。因此，关于癫痫对下丘脑Kispeptin的影响，人们的认识存在差距。 GnRH电路。这一具体建议的总体目标是阐明癫痫相关的变化。 Kispeptin-GnRH回路。在目标1中，将使用膜片钳电生理学来确定癫痫诱导的 Kispeptin神经元功能和兴奋性的变化与发情周期阶段和性别的关系。一种组合 将使用全细胞电流钳和松片记录来分析癫痫相关的变化 Kispeptin神经元固有的兴奋性和放电活动。在目标2中，将进行双光子钙成像 确定癫痫相关的GnRH神经元轴突终末活性的变化和对Kispeptin的反应。 完成这些目标将为继续取得进展提供必要的基础信息 控制癫痫相关疾病和预防癫痫的NINDS癫痫研究基准 癫痫治疗的不良后果。本提案中包含的培训计划旨在促进 发展首席调查员，为他作为独立研究员的职业生涯做好更好的准备。 这项培训计划非常强调以实验室为基础的学习，并增加少量额外的 还包括以课堂为基础的学习，为技术培训提供所需的概念基础。 这种研究体验也将在允许确定优先顺序的智力环境中进行 这些经验将在初级调查员的发展中发挥关键作用。

PROJECT SUMMARY Patients with temporal lobe epilepsy (TLE), the most commonly diagnosed form of focal epilepsy in adults, have been shown to experience reproductive endocrine disorders at a rate higher than the general population. Notably, clinical studies have also found disrupted patterns of luteinizing hormone (LH) release in both sexes. Gonadotropin-releasing hormone (GnRH) neurons form the final common output in the brain’s control of reproduction, and they play a key role in the pituitary release of LH. Our lab has shown that both GnRH neuron firing activity and intrinsic excitability are disrupted in the intrahippocampal kainic acid (IHKA) mouse model of TLE. Upstream kisspeptin neuron populations in the arcuate nucleus (ARC) and anteroventral periventricular nucleus (AVPV) are major sources of synaptic transmission to GnRH neurons. ARC kisspeptin neurons play a key role in the pulsatile release of LH in both sexes, and AVPV kisspeptin neurons regulate the preovulatory GnRH/LH surge in females. However, kisspeptin neuron function has never been studied in an animal model of epilepsy. Thus, there is a gap in knowledge regarding the effects of epilepsy on the hypothalamic kisspeptin- GnRH circuit. The overall objective of this specific proposal is to elucidate epilepsy-associated changes in the kisspeptin-GnRH circuit. In Aim 1, patch clamp electrophysiology will be employed to determine epilepsy-induced changes in kisspeptin neuron function and excitability in relation to estrous cycle stage and sex. A combination of whole-cell current clamp and loose-patch recordings will be used to analyze epilepsy-associated changes in kisspeptin neuron intrinsic excitability and firing activity. In Aim 2, two-photon calcium imaging will be conducted to identify epilepsy-associated changes in GnRH neuron axon terminal activity and response to kisspeptin. Completion of these aims will provide the foundational information necessary to make continued progress towards the NINDS Epilepsy Research Benchmarks of controlling epilepsy-related conditions and preventing the adverse consequences of seizure treatment. The training plan contained within this proposal seeks to facilitate the development of the primary investigator to better prepare him for a career as an independent researcher. This training plan places heavy emphasis on laboratory-based learning, with small amounts of additional classroom-based learning also included to provide the conceptual foundation needed for the technical training. This research experience will also take place in an intellectual environment that allows for the prioritization of those experiences that will play key roles in the development of the primary investigator.