Rapid seizure-induced changes in dendritic spines

描述（由申请人提供）：癫痫影响2 - 5%的人，通常与长期神经功能缺损有关，如学习障碍和脑瘫。虽然在某些临床情况下癫痫发作可能直接导致神经元死亡，但在其他情况下，癫痫发作似乎不会诱导神经元死亡，但仍可能对神经元结构和功能产生有害影响。虽然癫痫通过兴奋性毒性和细胞凋亡诱导的神经元死亡已被广泛研究，但癫痫发作引起的非致死性神经元损伤的机制知之甚少。最近，已经发现神经元的树突棘响应于各种生理刺激而表现出快速运动，这表明棘运动在重要的神经元功能（例如学习）中的作用。由于癫痫发作涉及过度的神经生理活动，本研究建议的主要假设是癫痫发作诱导树突棘的直接，快速变化，这取决于特定的癫痫发作特性。本研究的主要目的是利用先进的细胞成像技术，如共聚焦显微镜、双光子成像和表达绿色荧光蛋白的转基因小鼠，研究癫痫发作对动物癫痫模型树突棘密度、形态和运动性的快速影响。将在固定的脑切片中检查红藻氨酸盐和戊四唑诱导的体内癫痫发作对树突的时间过程和影响。在体内完整麻醉动物中，将分析树突状结构和运动性的实时变化。将在体外活体脑切片中研究细胞机制介导的快速脑损伤诱导的脊柱变化。主要研究者（PI）是一位在临床癫痫和基础癫痫研究方面接受过培训的学术医生-科学家。尽管PI在与动物癫痫发作模型相关的细胞和系统生理学方面具有丰富的经验，但本研究职业奖（K02）支持的高分辨率细胞成像的额外维度应该为癫痫发作引起的非致死性神经元损伤机制提供重要见解，并显着帮助PI发展癫痫的独立研究职业生涯。华盛顿大学在细胞成像的尖端技术和专家研究人员方面拥有无与伦比的资源，为PI完成本提案的目标提供了理想的环境。

DESCRIPTION (provided by applicant): Epilepsy affects 2-5% of people and is often associated with long-term neurological deficits, such as learning disabilities and cerebral palsy. Although seizures may directly cause neuronal death in some clinical contexts, in other situations seizures do not appear to induce neuronal death, but may still have detrimental effects on neuronal structure and function. While seizure-induced neuronal death via excitotoxicity and apoptosis has been studied extensively, mechanisms of non-lethal neuronal injury from seizures are poorly understood. Recently, dendritic spines of neurons have been found to exhibit rapid motility in response to various physiological stimuli, suggesting a role of spine motility in important neuronal functions, such as learning. As seizures involve excessive neurophysiological activity, the major hypothesis of this research proposal is that seizures induce direct, rapid changes in dendritic spines, which vary depending on specific seizure properties. The main objective of this proposal is to investigate the rapid effects of seizures on dendritic spine density, morphology and motility in animal seizure models, utilizing advanced cellular imaging techniques, such as confocal microscopy, two-photon imaging, and transgenic mice expressing green-fluorescent protein. The time course and effect of kainate and pentylenetetrazole-induced in vivo seizures on dendrites will be examined in fixed brain sections. Real-time changes in dendritic structure and motility will be analyzed during pharmacologically induced electrographic seizures in intact anesthetized animals in vivo. Cellular mechanisms mediating rapid seizure-induced spine changes will be investigated in live brain slices in vitro. The principal investigator (PI) is an academic physician-scientist trained in both clinical epilepsy and basic epilepsy research. Although the PI has substantial experience in cellular and systems physiology related to animal seizure models, the added dimension of high-resolution cellular imaging supported by this Research Career Award (K02) should provide important insights into mechanisms of non-lethal neuronal injury from seizures and significantly aid the PI in developing an independent research career in epilepsy. Washington University possesses unmatched resources in cutting-edge technology and expert researchers in cellular imaging and represents an ideal environment for the PI to complete the goals of this proposal.