Figure 2. Acute stress promotes neurite growth via a GC-GR-dependent mechanism. A, B, DRG neurons were isolated from ns or stressed mice and grown in vitro for 72 h. Scale bar, 100 µm. C, Stress increased neurite length. ***p < 0.0001 versus ns, one-way ANOVA. D-F, Stress increases neurite growth during the “arborizing” phase of DRG neurite outgrowth. D, DRG neurons from Thy1-GFP-M transgenic mice were isolated from ns adult mice or adult mice subjected to 1 h of restraint stress and grown for 15 h in vitro. Scale bar, 100 µm. E, F, Stress increased neurite total length and the number of branches per neurite, *p < 0.05, ***p < 0.0001, mean and SEM are shown. G, The percentage of neurons extending neurites was unaffected by stress. All parameters were defined by the Neuronal Profiling Algorithm v4.1 (Cellomics ArrayScanXTI, Thermo Fisher). N ≥ 550 neurons per condition. H, I, Cort administration (1.5 mg/kg) 1 h before DRG harvest increased neurite elongation and sprouting measures 24 h after plating in vitro. I, The synthetic GC, dex (2 mg/kg) increased neurite sprouting, *p < 0.05, **p < 0.01. J, Representative images of high-density DRG neuron cultures (72 h in vitro) from vehicle-treated and RU486-treated (RU) stressed mice. Scale bar, 200 µm. K, The GR antagonist, RU486 blocked stress enhanced neurite growth, ***p < 0.0001 versus ns, mean ± SEM are shown.