A caged neuron multielectrode system

该项目的目的是创建一个用于研究培养的神经网络的系统，该系统将提供一种独特的新能力，用于观察每个细胞如何与其他细胞连接以及连接性如何变化。 培养的神经元将在“笼子”中，“笼子”是允许过程生长但保持细胞体靠近记录和刺激电极的塑料结构。 通过刺激每个神经元并记录所有其他神经元的反应，网络的连通性将被揭示。 该系统将有64个笼在一个8 × 8的阵列，64个解离的神经元放置在笼使用光镊。 在单个小区级别详细研究这种网络的能力将 远远超过现在存在的任何东西。 记录和刺激不会损伤神经元，因此随着网络的发展，可以在几周内观察到连接。 将使用定制的大规模集成电路上的非常复杂的电子放大器和刺激器，这将以低成本提供同时研究多个笼状阵列的能力，同时它们在培养箱中。 这些研究将提供一个机会，以观察正常的网络发展的细节，这是从来没有可能的。 此外，强加的模拟对网络的影响将在几小时到几周的时间尺度上显示出来。 还将观察到对连接性的药理作用。 最后，因为它们在笼子里，所以细胞类型可以混合并放置在已知的位置，这样当它们形成网络时，它们之间的类型特异性相互作用将是可观察的。 一个最有趣的例子是观察胚胎干细胞衍生的神经元与正常神经元网络的相互作用。 作为本研究结束时的概念验证，将开始进行此类实验。 它们将表明干细胞-神经元在移植中与正常细胞整合的潜力，也将为研究如何促进这种整合提供机会。 公共卫生相关性：该项目提供的研究培养网络的能力将为网络发育，可塑性和药理学反应提供重要的新数据。 这将与使用慢性刺激和药物改善神经问题的医疗应用相关。 此外，胚胎干细胞神经元的实验将为研究神经移植与正常细胞的整合提供独特的机会，这与移植的可能医学应用非常相关。

DESCRIPTION (provided by applicant): The aim of this project is to create a system for studying cultured neural networks that will provide a unique new ability for observing how each cell is connected to the others and how that connectivity changes. The cultured neurons will be in "cages" that are plastic structures which allow process outgrowth but maintain the cell body in close proximity to a recording and stimulating electrode. By stimulating each neuron and recording the responses of all the others, the connectivity of the network will be revealed. The system will have 64 cages in an 8 x 8 array, with 64 dissociated neurons placed in cages using an optical tweezers. The capability to study such networks in detail at the single cell level will be far beyond any that now exists. Recording and stimulation do not damage the neurons, so the connectivity can be observed over a period of weeks, as the network develops. Very sophisticated electronic amplifiers and stimulators on custom large scale integrated circuits will be used, which will provide at low cost the capability of studying multiple caged arrays simultaneously, while they are in an incubator. The studies will provide an opportunity to observe normal network development in detail that has never been possible. In addition, the effects on networks of imposed simulation will be revealed, on time scales of hours to weeks. Pharmacological effects on connectivity will also be observable. Lastly, because they are in cages, cell types can be mixed and placed in known locations, so that type-specific interactions between them as they form networks will be observable. A most interesting example will be to observe the interaction of embryonic-stem-cell derived neurons with networks of normal neurons. As a proof of concept at the end of this study, such experiments will be begun. They will indicate the potential for stem-cell-neuron integration with normal cells in transplants, and also will provide opportunities for studying how to promote that integration. PUBLIC HEALTH RELEVANCE: The capability for studying cultured networks that this project provides will provide important new data on network development, plasticity, and pharmacological responses. This will be relevant to medical applications that ameliorate neurological problems using chronic stimulation and drugs. In addition, the experiments with embryonic stem cell neurons will provide a unique opportunity to study the integration of neural transplants with normal cells, which is very relevant to possible medical applications of transplants.