Micropatterned substrates coated with neuronal adhesion molecules for high-content study of synapse formation
Micropatterned substrates coated with neuronal adhesion molecules for high-content study of synapse formation
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DOI：
10.1038/ncomms3252
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发表时间：
2013-08-01
影响因子：
16.6
通讯作者：
Thoumine, Olivier
中科院分区：
文献类型：
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作者：
Czoendoer, Katalin;Garcia, Mikael;Thoumine, Olivier
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阻碍了对不同蛋白质的作用和突触发生机制的研究。在这里，我们展示了一个强大的和高内容的方法来诱导选择性突触前或突触后结构在控制的位置。在微图案化基底上培养神经元，所述微图案化基底包括涂覆有各种突触发生粘附分子的微米级点的阵列。当接种在neurexin-1 β包被的微图案上时，表达神经连接素-1的神经元表现出特异性树突组织和突触后支架分子PSD-95的选择性募集。此外，功能性AMPA受体被困在neurexin-1 β点，如通过实时成像实验所揭示的。相比之下，在SynCAM 1包被的基底上的神经元表现出强烈的模式化轴突和选择性组装功能性突触前体。然而，N-钙粘蛋白涂层不能引起突触，表明我们的系统的特异性。这种方法为各种突触系统的基础和治疗研究开辟了道路。
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-1 beta-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-1 beta 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.