EPILEPTIFORM ACTIVITY ON HIPPOCAMPAL MICROCULTURES

有人建议对突触进行直接的电生理研究 通过海马神经元的培养，重点研究突触参与的机制 得了癫痫。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，用来培养中央 神经系统神经元。这些微培养物可以包含最少一个 神经元，并且可以存活长达几个月。另一个进步是 Furshpanand Potter(1988)的体外癫痫模型 长时间生长的分离的海马神经元 在突触活动阻滞剂(如犬尿激素)存在的情况下进行培养 酸和镁)。当文化恢复正常生长时 介质中，神经元产生强烈的电活动爆发， 与完整的海马体记录的癫痫发作活动非常相似。 该项目的目标是： -研究海马区显微培养中的突触。微培养是一种 一种实用的生理学和药理学研究方法 突触相互作用，具有一些显著的优势 实验模型，如大脑皮层切片。神经元及其电路 将通过收集有关自燃的信息进行分类 模式、突触输入和突触输出，以及离子导入 回应。 -简化大众培养中的癫痫样活动模型 数以千计的神经元减少到微型培养中较少的神经元和 微培养，以研究癫痫现象的水平。 仅由几个神经元组成的系统中的单个突触。这个 将对长期被阻止的培养的异常进行调查 确定该模型系统中癫痫样活动的原因。这个 模型系统将被用来研究药物治疗 针对特定神经元亚群的癫痫，以避免 目前的癫痫治疗方法。

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.