Fig. 5: Optogenetic stimulation of VP Glut neurons drives avoidance, activates VTA GABA, DA and Glut neurons.

A Design of experiment with optostimulation of VP Glut neurons and simultaneous recording of GCaMP fluorescence in VTA DA (n = 6 mice, blue), Glut (n = 7 mice, green), or GABA neurons (n = 6 mice, red). B Time spent in side A during real-time place preference (RTPP) assay with blue shading representing stimulated side; fiber photometry (FP) recordings were made during the last two sessions. C Schematic illustrating passive stimulation and RTPP assays. D GCaMP responses of VTA DA, E VTA Glut, or F VTA GABA neurons during the 1-s or 5-s passive stimulation (shaded in blue, onset at t = 0 s). Histograms show mean peak z-score per animal during 1-s specified periods (pre-stimulation and stimulation for the 1-s stimulation; pre-, early- and late-stimulation for the 5-s stimulation assay). G GCaMP responses of VTA DA, H VTA Glut, or I VTA GABA neurons in response to stimulation induced by entry into the active side during RTPP (t = 0 s); only RTPP entries sustained for ≥1 s in the stimulation side (blue shading, fades after 1 s) and preceded by ≥2 s in the no-stimulation side (gray shading) were included. Histograms show mean peak z-score per animal during 1-s specified periods (pre-stimulation and stimulation). Statistics on histograms are results of t-test for D-F (left) & G-I, and of Tukey post-hoc following one-way ANOVAs for D-F (right). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by Tukey post-hoc following one-way ANOVA; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001, ^###p < 0.0001 by two-tailed paired t^-test. Data are represented as mean ± SEM. See Tables S1 and S2, also Fig. S1, S3, S7 and S8. Source data are provided as a Source Data file.