Abstract
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.
Highlights
Our study focuses on characterization of dorsal root ganglion (DRG) neurons cultured on silicon micropillar substrates (MPS) with the ultimate goal of designing micro-electrode arrays (MEAs) for successful electrophysiological recordings of Dorsal root ganglion (DRG) neurons
Dorsal root ganglion (DRG) neurons are an important site for pathophysiological changes that lead to neuropathic pain[1]
With advanced MEAs based on integrated complementary metal oxide semiconductor (CMOS) any neuron grown over custom arrays can be recorded at high spatio-temporal resolution allowing us to better understand some aspects of its altered electrophysiological properties[7]
Summary
Our study focuses on characterization of dorsal root ganglion (DRG) neurons cultured on silicon micropillar substrates (MPS) with the ultimate goal of designing micro-electrode arrays (MEAs) for successful electrophysiological recordings of DRG neurons. 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. 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. In order to use MEAs for a wide range of neuroscience applications, including neuro-regenerative medicine, construction of neural networks and electrophysiology studies, it is crucial to design and produce micro-pillar substrates (MPS) with specific topography that will provide favorable growth and desirable morphology of cultured neurons or precise guidance and positioning of their neurites[9,10,11,12]. We focused on the influence of MPS isotropic features on presence and morphology, i.e. neurite alignment, length www.nature.com/scientificreports/
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