Journal: Frontiers in Neuroscience

Article Title: Human sensory-like neuron cultivation—An optimized protocol

doi: 10.3389/fnins.2024.1429694

Figure Lengend Snippet: Validation of neuronal integrity. (A) Viability assay of iSNs following treatment with increasing concentrations of FdU and AraC for up to 72 h. SDS was used as negative ctrl, 10 μM of FdU for 24 h. The data of n = 3 differentiations, one-way ANOVA with post-hoc Tukey's HSD. * p < 0.05; *** p < 0.001. (B) iSNs generated using our optimized protocol express the sensory neuron marker Nav1.8, the peripheral neuron marker PRPH, and a general neuronal marker TUJ1. The scale bar = 100 μm. (C, D) Spontaneous electrical activity in iSNs was measured at 4 (I), 5 (II), and 6 (III) weeks of maturation using a multi-electrode array system, with activity detectable starting from 4 weeks of maturation. Scale: 50 μV/10 s. (D) Raster plot of one well over 4 (I), 5 (II), and 6 (III) weeks of maturation showing the single and developing network activity. For (C, D) n = 3 triplicates from 1 differentiation. ICC, immunocytochemistry; Nav1.8, voltage-gated sodium channel 1.8; PRPH, peripherin; TUJ1, Beta-III-tubulin.

Article Snippet: The electrical activity of iSNs was assessed using a multiwell multi-electrode array (MEA) system comprising 24 individual wells with 12 electrodes per well (Multi Channel Systems MCS GmbH, Reutlingen, Germany).

Techniques: Viability Assay, Generated, Marker, Activity Assay, Immunocytochemistry

Journal: Cell Death & Disease

Article Title: Tau P301L disengages from the proteosome core complex and neurogranin coincident with enhanced neuronal network excitability

doi: 10.1038/s41419-024-06815-2

Figure Lengend Snippet: All experiments were undertaken in DIV12 cultures cultured in CytoView micro-electrode array (MEA) multi-well plates using the Maestro-Pro MEA system (Axion Biosystems). A Each well contains 16 electrodes that can record local activity B Fluorescent microscopy shows DIV12 primary cortical neurons transduced with AAV-Syn1-eGFP and cultured on a micro-electrode plate. Scale bar = 275 µm. C An example of a raster plot, which shows firing over time, taken at baseline from untransduced neurons. Each row corresponds to an individual electrode. Action potentials are recorded as spikes (black lines) and this is termed ‘firing’. A burst (blue) occurs when multiples action potentials fire in rapid succession in the locality of an electrode; a minimum threshold is set at 50 spikes within 100 ms. A network burst was classified as at least 35% of electrodes recording bursts simultaneously; this shows that the neurons throughout the culture are connected. D Representative raster plots showing 60 s recordings of untransduced neurons at baseline and following treatment with 0.625 µM 4AP/12.5 µM Bicuculline (Bic). Baseline recordings show spontaneous firing and bursting. Treatment with 4AP/Bic stimulates synchronous and oscillatory firing across the well, as shown by a switch to network bursting. E 4AP/Bic treatment significantly increases activity and ( F ) burst strength ( t -test, **** P < 0.0001, n = 3 technical repeats). G Representative raster plots comparing neurons overexpressing eGFP, eGFP-Tau WT or eGFP-Tau P301L at baseline and following treatment with 4AP/Bic. H At baseline, the activity and ( I ) burst strength of neurons overexpressing eGFP-Tau P301L is significantly greater than those expressing eGFP or eGFP-Tau WT . Following treatment with 4AP/Bic, activity but not burst strength is significantly greater with eGFP-Tau P301L than eGFP or eGFP-Tau WT (two-way ANOVA with Tukey’s multiple comparisons test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 4 technical repeats).

Article Snippet: All experiments were undertaken in DIV12 cultures cultured in CytoView micro-electrode array (MEA) multi-well plates using the Maestro-Pro MEA system (Axion Biosystems).

Techniques: Cell Culture, Activity Assay, Microscopy, Transduction, Expressing