Figure 5. Activation of Tacr1- and Gpr83-expressing SPB neurons induces distinct affective behaviors in a stimulus intensity-dependent manner.

a, Schematic of the optogenetic stimulation-coupled real-time place preference assay. b, c, Quantifications of % of time spent in stimulated side for 6.5 mW (b) and 1 mW (c) photostimulations (10 Hz, 10 ms pulse width). One-way repeated measures ANOVA (Dunnett’s multiple comparisons test); F_{[1.626, 8.131]} = 19.10 (Tacr1), F_{[1.580, 7.901]} = 14.41 (Gpr83) (b); F_{[1.903, 13.32]} = 7.20 (Tacr1), F_{[1.817, 12.72]} = 8.42 (Gpr83) (c); n = number of mice. d, Schematic of the optogenetic stimulation-coupled lever-pressing assay. Mice received 5 second-long photostimulation (473 nm, 10 Hz, 10 ms pulse width) upon pressing an active lever. e, f, Fold changes of number of lever press are plotted over sessions. Turquoise boxes, 8 days of light-on sessions. n = 7 (control, 0.4 mW; Tacr1, 0.4 mW; Gpr83, 0.4 mW; Tacr1, 1 mW), n = 6 (control, 1 mW), n = 8 mice (Gpr83, 1 mW). Two-way repeated measures ANOVA; F_{[1, 12]} = 5.26 (0.4 mW), F_{[1, 12]} = 5.22 (1 mW) (e); F_{[1, 12]} = 2.50 (0.4 mW), F_{[1, 11]} = 10.14 (1 mW). g, Distribution of Fos⁺ neurons in the PBN_EL following photostimulation of Gpr83⁺ SPB axon terminals. h-j, Quantifications of the number of Fos⁺ neurons in different PBN_L subnuclei (h), % of CGRP⁺ neurons that are Fos⁺ (i), and % of Fos⁺ neurons that are CGRP⁺ (j). One-way ANOVA (Tukey’s multiple comparisons test); F_{[2, 8]} = 7.41 (PBN_IL), F_{[2, 8]} = 22.03 (PBN_DL), F_{[2, 8]} = 64.36 (PBN_EL); F_{[2, 8]} = 74.94 (i); n = 4, 4, 3 mice for control, 0.4 mW, and 6.5 mW, respectively. Error bars, s.e.m.