EPILEPTIFORM ACTIVITY ON HIPPOCAMPAL MICROCULTURES

建议对突触进行直接的电生理研究， 海马神经元的培养，重点是突触机制， 在癫痫中。这项工作借鉴了我们小组的两项实验进展。一 是我开发的一种技术， 神经系统神经元这些微培养物可以包含少至一个 神经元，可以存活长达数月。另一个进步是 Furshpan和Potter（1988）的体外癫痫模型， 分离的海马神经元，其生长时间较长， 在突触活性阻断剂（如犬尿氨酸）存在下培养 酸和镁）。当培养物恢复正常生长时 当神经元处于中等水平时，神经元产生强烈的电活动爆发， 与完整海马体记录的癫痫发作活动非常相似。 该项目的目标是： - 研究海马微培养物中的突触。微培养是一种 用于生理学和药理学研究的有用的通用方法 突触的相互作用，与其他一些显着的优势， 实验模型，如皮质切片。神经元及其电路 将通过收集自发放电的信息进行分类 模式，突触输入和突触输出，以及离子电渗 应答 - 为了简化癫痫样活动的模型， 从数千个神经元到更小数量的神经元， 微培养，以研究癫痫现象的水平， 在一个仅由几个神经元组成的系统中的单个突触。的 将研究慢性阻塞培养物的异常， 确定该模型系统中癫痫样活动的原因。的 模型系统将用于研究药物治疗 针对特定神经元亚群的癫痫，以避免 癫痫病的治疗方法

It is proposed to do direct electro-physiological studies of synapses made by hippocampal neurons in culture, focusing on synaptic mechanisms involved in epilepsy. This work draws on two experimental advances of our group. One is a technique I have developed for growing "microcultures" of central nervous system neurons. These microcultures can contain as few as one neuron, and can survive for as long as several months. The other advance is an in-vitro epilepsy model of Furshpan and Potter (1988) that involves dissociated hippocampal neurons that are grown for prolonged periods in culture in the presence of blockers of synaptic activity (such as kynurenic acid and magnesium). When the cultures are returned to regular growth medium, the neurons produce intense bursts of electrical activity that closely resemble the seizure activity recorded from the intact hippocampus. Aims for the project are: - To study synapses in hippocampal microcultures. Microcultures are a useful general method for physiological and pharmacological studies of synaptic interactions, with some significant advantages over other experimental models such as cortical slices. Neurons and their circuitry will be categorized by collecting information about spontaneous firing patterns, synaptic inputs and synaptic outputs, and iontophoretic responses. - To simplify the model of epileptiform activity in mass-cultures of thousands of neurons down to smaller numbers of neurons in minicultures and microcultures, in order to study the phenomenon of epilepsy at the level of individual synapses in a system consisting of only several neurons. The abnormalities of the chronically blocked cultures will be investigated to determine the cause of the epileptiform activity in this model system. The model system will be used to investigate pharmacological treatments of epilepsy that are targeted at specific subgroups of neurons, to avoid current epilepsy treatments.