Figure 2.

Anxiety-like behavior and hindpaw sensitivity were dose-dependently affected by neonatal maternal separation (NMS) and vaginal balloon distension (VBD). The percent of time spent in the perimeter of an open field actimeter was recorded as a measure of behavioral anxiety in NMS14 (A), NMS21 (B), and corresponding naïve mice, both prior to (Baseline) and following VBD (Post-VBD). A) No significant effect of NMS or VBD on exploratory behavior was observed within the NMS14 group. B) NMS21 mice displayed a significant decrease in the percent of time spent in the perimeter of the open field at baseline compared to naïve mice. Following VBD, NMS21 mice spent a significantly larger percentage of their time in the perimeter compared to both post-VBD naïve mice and baseline NMS21 mice. C) The withdrawal latency of the hindpaw to a thermal stimulus was significantly shorter in NMS14 mice only following VBD, compared to both post-VBD naïve and NMS14 baseline measurements. D) NMS21 mice displayed a significantly shorter withdrawal latency to thermal stimulation of the hindpaw than naïve mice, both at baseline and post-VBD. E) The withdrawal threshold of the hindpaw to a mechanical stimulus was significantly lower in NMS14 mice only following VBD, compared to post-VBD naïve mice. F) NMS21 mice also displayed a significantly lower withdrawal threshold to mechanical stimulation of the hindpaw than naïve mice, both at baseline and post-VBD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 vs. naïve, ^##p < 0.01, ^####p < 0.0001 vs. baseline; two-way ANOVA and Bonferroni posttest.