Fig. 2: LHA glutamatergic neurons projecting to VTA are stress sensitive.
a Timeline and schematic for Fiber photometry experiments during stress. b Representative image of viral expression of GCaMP6s in LHAglu neurons projecting to the VTA (in green), with an optic fiber above the LHA (scale bars: 100 µm). Anatomical reference points: f fornix, EPN entopeduncular nucleus, mt mammillothalamic tract. c Average recorded emitted fluorescent signal of GCaMP6s with SEM (dF/F) (green; n = 10) and control GFP (black; n = 10) of LHAglu neurons projecting to VTA, time-locked to fights (red line). Scale bars: 2 s, 0.25 dF/F. Quantified peak signals compared to baseline during fight onset of GCaMP and GFP control groups with SEM (mice per group n = 10, One-Way ANOVA F(1,18) = 12.03, p = 0.003). d As (c), but for fight offset (end of the red line) (mice per group n = 10, One-Way ANOVA, F(1,18) = 3.65, p = 0.072). e Average recorded emitted fluorescent signal plus SEM of GCaMP6s signal and GFP (dF/F) of LHAglu-VTA neurons time-locked to the onset of high velocity bouts. Scale bars: 1 s, 0.25 dF/F. Quantified peak signals compared to baseline during high velocity bouts for GCaMP and GFP control groups. Error bars indicate SEMs (mice per group n = 9, One-Way ANOVA, F(1,16) = 1.00, p = 0.33). f Average recorded emitted fluorescent signal plus SEM of GCaMP6s signal and GFP (dF/F) of LHAglu neurons projecting to VTA time-locked to juvenile interactions. Scale bars: 1 s, 0.25 dF/F. Quantified peak signals compared to baseline during juvenile interaction for GCaMP and GFP control groups. Error bars indicate SEMs (mice per group n = 10, One-Way ANOVA, F(1,18) = 1.75, p = 0.20). g Timeline and schematic for fiber photometry experiments during nose pokes for fat. On the right same as (b). h Left: Average recorded emitted fluorescent signal of GCaMP6s plus SEM (dF/F) (pre and post control/stress, respectively black and blue; n = 4) and stress (pre and post stress, respectively black and red; n = 8) of LHAglu neurons projecting to VTA, time-locked to the onset of nose pokes for fat (dashed orange line). Scale bars: 1 s, 0.1 dF/F. Right: Quantified peak signals compared to pre nose poke during fat nose poke of control and stress animals pre and post stress +SEM (RM Two-way ANOVA. Main effect stress F(1,10) = 0.338, p = 0.54; main effect time F(1,10) = 0.004, p = 0.95; main interaction effect time*stress, F(1,10) = 0.535, p = 0.48) . All statistical tests were performed two-sided. **p < 0.01. Source data are provided as a Source Data file.
