Engineering connectivity by multiscale micropatterning of individual populations of neurons
Engineering connectivity by multiscale micropatterning of individual populations of neurons
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DOI：
10.1002/biot.201400609
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发表时间：
2015-02-01
影响因子：
4.7
通讯作者：
Offenhaeusser, Andreas
中科院分区：
文献类型：
--
作者：
Albers, Jonas;Toma, Koji;Offenhaeusser, Andreas
功能网络是中枢神经系统信息处理的基础。其形成的关键是指导神经元生长以及控制连接和信息流。神经元发育的基础是由引导线索和几何约束产生的。为了研究神经元的生长和相邻神经元网络的连接，创建了二维蛋白质模式。通过微接触印刷将聚-L-赖氨酸和层粘连蛋白的混合物转移到硅烷化玻璃表面上。这些结构中填充有分离的原代皮层胚胎大鼠神经元。选择具有不同开口角度、高度和设计的三角形结构作为二维结构，以允许具有收缩的网关的网络形成。通过免疫组织化学观察神经元的发育，以追踪所选结构对神经元生长的影响。对神经元进行MAP 2染色，而聚-L-赖氨酸被FITC标记。通过这项研究，我们提出了一种易于使用的技术，工程师在体外二维网络与定义的网关。所提出的微图案化方法用于生成具有预定义连接的菊花链神经元网络。几何约束网络之间的信号传播可以很容易地监测钙敏感染料，提供洞察网络通信在体外。
Functional networks are the basis of information processing in the central nervous system. Essential for their formation are guided neuronal growth as well as controlled connectivity and information flow. The basis of neuronal development is generated by guiding cues and geometric constraints. To investigate the neuronal growth and connectivity of adjacent neuronal networks, two-dimensional protein patterns were created. A mixture of poly-L-lysine and laminin was transferred onto a silanized glass surface by microcontact printing. The structures were populated with dissociated primary cortical embryonic rat neurons. Triangular structures with diverse opening angles, height, and design were chosen as two-dimensional structures to allow network formation with constricted gateways. Neuronal development was observed by immunohistochemistry to pursue the influence of the chosen structures on the neuronal outgrowth. Neurons were stained for MAP2, while poly-L-lysine was FITC labeled. With this study we present an easy-to-use technique to engineer two-dimensional networks in vitro with defined gateways. The presented micropatterning method is used to generate daisy-chained neuronal networks with predefined connectivity. Signal propagation among geometrically constrained networks can easily be monitored by calcium-sensitive dyes, providing insights into network communication in vitro.