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. 2021 Mar 5;10:e60779. doi: 10.7554/eLife.60779

Figure 6. Photostimulation of PdynPBN→POA, PenkPBN→POA, and VGLUT2PBN→POA terminals induces real time place aversion.

(A, D, and G) Illustrations of viral injections in parabrachial nucleus (PBN) and fiber optic implantations over POA in Pdyn-Cre mice, Penk-Cre, and VGLUT2-Cre mice, respectively. (B, E, and H) Representative heat maps showing spatial distribution of time-spent behavior resulting from side-conditional 10 Hz photostimulation of control or Pdyn-Cre, Penk-Cre, and VGLUT2-Cre mice, respectively. (C) For Pdyn-Cre vs control mice, frequency response of RTPP at 0 (baseline), 2, 5, 10, and 20 Hz. Data are presented as mean ± SEM; n = 6 Cre+, eight control; two-Way ANOVA, Bonferroni post hoc. (F) Penk-Cre frequency response of RTPP at 0 (baseline), 2, 5, 10, and 20 Hz. Data are presented as mean ± SEM; n = 6 Cre+, seven control; two-Way ANOVA, Bonferroni post hoc (5 Hz ChR2 vs. 5 Hz control ∗∗∗p<0.001, 10 Hz ChR2 vs. 10 Hz control ∗∗∗∗p<0.0001, 20 Hz ChR2 vs. 20 Hz control ∗∗∗∗p<0.0001). (I) VGLUT2-Cre frequency response of RTPP at 0 (baseline), 2, 5, 10, and 20 Hz. Data are presented as mean ± SEM; n = 8 Cre+, seven control; two-Way ANOVA, Bonferroni post hoc (2 Hz ChR2 vs. 20 Hz control ∗∗∗∗p<0.0001, 5 Hz ChR2 vs. 5 Hz control ∗∗∗∗p<0.0001, 10 Hz ChR2 vs. 10 Hz control ∗∗∗∗p<0.0001, 20 Hz ChR2 vs. 20 Hz control ∗∗∗∗p<0.0001). See also Figure 6—figure supplement 1.

Figure 6.

Figure 6—figure supplement 1. Total distance traveled for Pdyn-Cre, Penk-Cre, and VGLUT2-Cre mice in real-time place aversion assay and male vs. female photostimulation-induced body temperature change in Pdyn-Cre and VGLUT2-Cre mice.

Figure 6—figure supplement 1.

(A) Distance moved during RTPP trials displayed in Figure 6C (Pdyn-Cre). Data are presented as mean ± SEM; n = 6 Cre+, eight control; two-way ANOVA, Bonferroni post hoc (20 Hz ChR2 vs. 20 Hz control ∗∗p<0.01). (B) Distance moved during RTPP trials displayed in Figure 6F (Penk-Cre). Data are presented as mean ± SEM; n = 6 Cre+, seven control; two-Way ANOVA, Bonferroni post hoc (not significant at all frequencies ns p>0.05). (C) Distance moved during RTPP trials displayed in Figure 6I (VGLUT2-Cre). Data are presented as mean ± SEM; n = 8 Cre+, seven control; two-Way ANOVA, Bonferroni post hoc (not significant at all frequencies ns p>0.05). (D and E) The change in body temperature of Pdyn-Cre female (n = 4) and male (n = 2) and VGLUT2-Cre female (n = 6) and male (n = 2) mice was similar at all stimulation frequencies (2, 5, 10, and 15 Hz) tested.