Regulatory effects of gradient microtopographies on synapse formation and neurite growth in hippocampal neurons
Regulatory effects of gradient microtopographies on synapse formation and neurite growth in hippocampal neurons
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DOI：
10.1088/1361-6439/ac73d7
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发表时间：
2022-05
影响因子：
2.3
通讯作者：
R. McNaughton;Yuda Huo;Guicai Li;Anaïs Di Via Ioschpe;Lei Yan;H. Man;Xin Zhang
中科院分区：
文献类型：
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作者：
R. McNaughton;Yuda Huo;Guicai Li;Anaïs Di Via Ioschpe;Lei Yan;H. Man;Xin Zhang
在旨在恢复功能性神经系统的方法中，实施微制造技术的方法允许制造具有不同几何形状的平台，其中神经元可以在体外发育并被引导形成图案化连接。神经元发育和由物理限制形成的微环境之间的相互作用在很大程度上仍然未知。因此，至关重要的是要有一个有效的方法来量化的物理或接触的指导微环境诱导的神经元形态学变化。在这项研究中，我们首先设计和评估的方法来制备各向异性，梯度聚（二甲基硅氧烷）微脊/槽阵列具有可变的局部图案宽度。然后，我们证明了这种单一基板的能力，同时配置文件的原代培养海马神经元的形态和突触连接的变化，在整个神经发育，在体外可变的物理conditons。梯度微形貌增强了微槽内的粘附力，随着图案宽度的减小，索马密度增加。减少图案宽度也减少了树突状分支和增加优先轴突生长。最后，减少图案的几何形状抑制突触前点的架构。总的来说，建立了一种方法来检查响应于物理刺激的结构发展和连接，并可能提供深入了解微加工的几何形状，促进神经再生和修复。
Among approaches aiming toward functional nervous system restoration, those implementing microfabrication techniques allow the manufacture of platforms with distinct geometry where neurons can develop and be guided to form patterned connections in vitro. The interplay between neuronal development and the microenvironment, shaped by the physical limitations, remains largely unknown. Therefore, it is crucial to have an efficient way to quantify neuronal morphological changes induced by physical or contact guidance of the microenvironment. In this study, we first devise and assess a method to prepare anisotropic, gradient poly(dimethylsiloxane) micro-ridge/groove arrays featuring variable local pattern width. We then demonstrate the ability of this single substrate to simultaneously profile the morphologcial and synaptic connectivity changes of primary cultured hippocampal neurons reacting to variable physical conditons, throughout neurodevelopment, in vitro. The gradient microtopography enhanced adhesion within microgrooves, increasing soma density with decreasing pattern width. Decreasing pattern width also reduced dendritic arborization and increased preferential axon growth. Finally, decreasing pattern geometry inhibited presynaptic puncta architecture. Collectively, a method to examine structural development and connectivity in response to physical stimuli is established, and potentially provides insight into microfabricated geometries which promote neural regeneration and repair.