Laboratory for Pain Research, School of Medicine, University of Split, Croatia.
Speech and Hearing Research Laboratory, School of Medicine, University of Split, Croatia.
Sci Rep. 2016 Dec 23;6:39560. doi: 10.1038/srep39560.
Our study focuses on characterization of dorsal root ganglion (DRG) neurons cultured on silicon micro-pillar substrates (MPS) with the ultimate goal of designing micro-electrode arrays (MEAs) for successful electrophysiological recordings of DRG neurons. Adult and neonatal DRG neurons were cultured on MPS and glass coverslips for 7 days in vitro. DRG neuronal distribution and morphometric analysis, including neurite alignment and length, was performed on MPS areas with different pillar width and spacing. We showed that MPS provide an environment for growth of adult and neonatal DRG neurons as permissive as control glass surfaces. Neonatal DRG neurons were present on MPS areas with narrow pillar spacing, while adult neurons preferred wider pillar spacing. Compared to the control glass surfaces the neonatal and adult DRG neurons in regions with narrow pillar spacing range developed a smaller number of longer neurites. In the same area, neurites were preferentially oriented along three directional axes at 30°, 90° and 150°. MPS architecture influenced growth directionality of all main DRG neuronal subtypes. We can conclude that specific micro-pillar substrate topography affects the morphology of DRG neurons. This knowledge can enable development of MEAs with precisely defined physical features for various neuroscience applications.
我们的研究集中在培养背根神经节(DRG)神经元上,这些神经元培养在硅微柱基底(MPS)上,最终目的是设计用于成功记录 DRG 神经元电生理活动的微电极阵列(MEA)。成年和新生 DRG 神经元在 MPS 和玻璃盖玻片上培养 7 天。在具有不同柱宽和柱间距的 MPS 区域上进行了 DRG 神经元分布和形态计量分析,包括神经突的排列和长度。我们表明,MPS 为成年和新生 DRG 神经元的生长提供了一个与对照玻璃表面一样有利的环境。新生 DRG 神经元存在于柱间距较窄的 MPS 区域,而成年神经元则更喜欢较宽的柱间距。与对照玻璃表面相比,在柱间距较窄的区域,新生和成年 DRG 神经元发育出的长神经突数量较少。在同一区域,神经突优先沿 30°、90°和 150°三个方向轴定向。MPS 结构影响所有主要 DRG 神经元亚型的生长方向。我们可以得出结论,特定的微柱基底形貌会影响 DRG 神经元的形态。这一知识可以为各种神经科学应用开发具有精确定义物理特征的 MEA。