Laboratoire de Neurobiologie des Processus Adaptatifs, CNRS, UMR7102, Université Pierre et Marie Curie, Paris, France.
Lab Chip. 2011 Nov 7;11(21):3663-73. doi: 10.1039/c1lc20014c. Epub 2011 Sep 15.
Various experimental models are used to study brain development and degeneration. They range from whole animal models, which preserve anatomical structures but strongly limit investigations at the cellular level, to dissociated cell culture systems that allow detailed observation of cell phenotypes but lack the highly ordered physiological neuron connection architecture. We describe here a platform comprising independent cell culture chambers separated by an array of "axonal diodes". This array involves asymmetric micro-channels, imposing unidirectional axon connectivity with 97% selectivity. It allows the construction of complex, oriented neuronal networks not feasible with earlier platforms. Different neuronal subtypes could be co-cultivated for weeks, and sequential seeding of different cell populations reproduced physiological network development. To illustrate possible applications, we created and characterized a cortico-striatal oriented network. Functional synaptic connections were established. The activation of striatal differentiation by cortical axons, and the synchronization of neural activity were demonstrated. Each neuronal population and subcompartment could be chemically addressed individually. The directionality of neural pathways being a key feature of the nervous system organization, the axon diode concept brings in a paradigmatic change in neuronal culture platforms, with potential applications for studying neuronal development, synaptic transmission and neurodegenerative disorder such as Alzheimer and Parkinson diseases at the sub-cellular, cellular and network levels.
各种实验模型被用于研究大脑发育和退化。它们的范围从保留解剖结构但强烈限制细胞水平研究的整体动物模型，到允许详细观察细胞表型但缺乏高度有序的生理神经元连接结构的分离细胞培养系统。我们在这里描述了一个由独立的细胞培养室组成的平台，这些培养室由一系列“轴突二极管”隔开。该阵列涉及非对称微通道，对轴突连接施加单向性，具有 97%的选择性。它允许构建以前的平台难以实现的复杂、定向的神经元网络。不同的神经元亚型可以共同培养数周，并且不同细胞群体的顺序播种可以再现生理网络的发育。为了说明可能的应用，我们创建并表征了一个皮质纹状体定向网络。建立了功能性突触连接。皮层轴突对纹状体分化的激活和神经活动的同步性得到了证明。每个神经元群体和亚区都可以单独进行化学处理。神经通路的方向性是神经系统组织的一个关键特征，轴突二极管的概念在神经元培养平台中带来了范式转变，有可能用于在亚细胞、细胞和网络水平上研究神经元发育、突触传递和阿尔茨海默病和帕金森病等神经退行性疾病。
