Fig. 3
From: Acid Sensing Ion Channels (ASICs) in NS20Y cells – potential role in neuronal differentiation
Effect of PcTX1 on cpt-cAMP induced differentiation of NS20Y cells. a Example images showing morphology of NS20Y cells in different treatment groups. For Sholl analysis of dendritic complexity, concentric rings were placed at the center of the soma radiating outward at a 10 μm intervals. Neurite complexity is measured by the number of neurites per intersection. Top panel: Control - shows NS20Y cells grown in media without pharmacologic treatment. Cells display a full, rounded soma with few, short undeveloped processes. Middle panel: cpt-cAMP treated - shows NS20Y cells treated with 1 mM cpt-cAMP for 72-h. Cells have large cell bodies, multiple and elongated processes with extensive branching. Lower panel: cpt-cAMP + PcTX1 - shows NS20Y cells treated with 1 mM cpt-cAMP and 10 nM PcTX1 for 72-h. These cells have a reduced neurite extension and processes. Photomicrographs were taken at 400×, phase contrast. b Summary data expressed as the mean neurite length/cell at time points of 24, 48, and 72 h. cpt-cAMP significantly increases the mean length of neurite extension (# p < 0.05 vs. control). When PcTX1 is added, there is a significant reduction in neurite length compared to cpt-cAMP treated cells (* p < 0.05). c Summary Sholl analysis data of control, cpt-cAMP (1 mM) treated and cpt-cAMP (1 mM) + PcTX1 (10nM) treated cells at 72 h. The plot shows the mean number of neurite intersections at indicated distance from the soma. Treatment with cpt-cAMP significantly increased the dendritic complexity (p < 0.05 vs. control, two way ANOVA, n = 90 cells for each group from 3 separate experiments). Co-treatment with 10 nM PcTX1 significantly attenuated the increase of dendritic complexity induced by cpt-cAMP (p < 0.05 between cpt-cAMP and cpt-cAMP + PcTX1, two way ANOVA, n = 90 cells from 3 separate experiments)