University of Bordeaux, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux, France.
Nat Commun. 2013;4:2252. doi: 10.1038/ncomms3252.
Studying the roles of different proteins and the mechanisms involved in synaptogenesis is hindered by the complexity and heterogeneity of synapse types, and by the spatial and temporal unpredictability of spontaneous synapse formation. Here we demonstrate a robust and high-content method to induce selectively presynaptic or postsynaptic structures at controlled locations. Neurons are cultured on micropatterned substrates comprising arrays of micron-scale dots coated with various synaptogenic adhesion molecules. When plated on neurexin-1β-coated micropatterns, neurons expressing neuroligin-1 exhibit specific dendritic organization and selective recruitment of the postsynaptic scaffolding molecule PSD-95. Furthermore, functional AMPA receptors are trapped at neurexin-1β dots, as revealed by live-imaging experiments. In contrast, neurons plated on SynCAM1-coated substrates exhibit strongly patterned axons and selectively assemble functional presynapses. N-cadherin coating, however, is not able to elicit synapses, indicating the specificity of our system. This method opens the way to both fundamental and therapeutic studies of various synaptic systems.
研究不同蛋白质在突触发生中的作用及其相关机制受到突触类型的复杂性和异质性以及自发突触形成的空间和时间不可预测性的阻碍。在这里，我们展示了一种强大且高内涵的方法，可选择性地在控制位置诱导突触前或突触后结构。神经元在微图案化的基板上培养，该基板包含涂有各种突触形成粘附分子的微米级点的阵列。当在神经连接蛋白 1β 涂覆的微图案上培养时，表达神经粘连蛋白 1 的神经元表现出特定的树突组织和突触后支架分子 PSD-95 的选择性募集。此外，通过实时成像实验揭示了功能 AMPA 受体被困在神经连接蛋白 1β 点上。相比之下，在 SynCAM1 涂覆的基板上培养的神经元表现出强烈的模式化轴突，并选择性地组装功能前突触。然而，N-钙粘蛋白涂层不能引发突触，表明我们的系统具有特异性。这种方法为各种突触系统的基础和治疗研究开辟了道路。
