Figure 3
Progression of BNSTCRF Activity Across Noxious Heat Exposure Trials. (A) Average BNSTCRF traces of male (purple; n = 49 cells / 7–30 cells per mouse / 3 mice total) and female (magenta; n = 68 cells / 8–19 cells per mouse / 5 mice total) subjects by trial. Within each representative time window, the maximum heat duration (20 s) is indicated in the transparent blue block. (B-C) Percentage of pain responsive BNSTCRF neurons for male (purple) and female (magenta) subjects were determined relative to epochs surrounding the onset of (B) heat exposure [“Relative to Before”] and (C) paw withdrawal [“Relative to After”] in trials 1–4. Positive, negative, and non-responsive cells are indicated by darker purple/magenta, lighter purple/magenta, and grey respectively (Wilcoxon rank-sum, P < 0.05). Comparisons by trial and sex relative to the epochs surrounding heat onset (Two-way mixed-model ANOVA with Sidak’s post hoc: no Trial × Sex interaction [F(3, 18) = 0.0466, P = 0.9862] or main effect of Trial [F(1.092, 6.553) = 0.4473, P = 0.5435] and Sex [F(1, 6) = 1.510, P = 0.2651]) and paw withdrawal (Two-way mixed-model ANOVA with Sidak’s post hoc: no Trial × Sex interaction [F(3, 18) = 0.3670, P = 0.7777] or main effect of Trial [F(2.740, 16.44) = 0.3368, P = 0.7818] and Sex [F(1, 6) = 0.0008, P = 0.9776]) were not statistically significant. (D) Heatmap displaying the average z-score of individual BNSTCRF neurons by epoch for male and female subjects in trials 1–4. Epochs are color-coded: Before (blue), Heat Start (red), Heat End (orange), and After (green), with darker shades of color corresponding to greater changes in Ca2+ activity. Data are shown as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001, ****P < 0.0001.
