Fig. 2: Peptidergic PBN subpopulations have opposing effects on breathing.

a Example RNAscope images illustrating partially overlapping Tac1, Calca, and Oprm1 populations in the PBL (a₁) with Venn Diagram (a₂) and table (a₃) showing the estimated size of each subpopulation as a fraction of total PBL cells (n = 9839 cells analyzed from n = 3 mice). Green arrows in a₁ and green box in a₃ indicate Tac1 neurons that do not co-express Calca. Scale bars = 40 µm and 20 µm (zoomed). b Schematic of viral gene transfer approach (b₁) and recombinase-dependent expression of ChR2:YFP in Tac1 (b₂) or CGRP (b₃) PBN neurons representative of n = 7 (Tac1) or n = 6 (CGRP) mice. Scale bar = 200 µm. scp superior cerebellar peduncle, Cbx cerebellar cortex. c Representative plethysmograph recordings from a mouse when awake (top) and lightly anesthetized with 1.5% isoflurane (bottom) during photoactivation of PBN^Tac1 neurons for 15 s at 30 Hz. Panels on right show overlay of 1 s of breathing with light OFF (black) and light ON (blue). d Same as c but during photoactivation of PBN^CGRP neurons. Scale bars in c also apply to d. e PCA comparing spontaneous breathing patterns (22,351 breaths from n = 6 mice) to breathing patterns during photoactivation of PBN^Tac1 (3,507 breaths from n = 7 mice) and PBN^CGRP (848 breaths from n = 6 mice) neurons. f Probability density histograms (top) and mean values from each mouse (bottom; n = 7 Tac1, n = 6 CGRP) comparing breathing frequencies and peak inspiratory pressures during photoactivation of Tac1 or CGRP PBN neurons. Tac1 and CGRP data were pooled for the light OFF condition shown in the probability density histograms. Light OFF and ON conditions and Tac1 vs. CGRP Light ON conditions were compared using two-tailed Wilcoxon matched pairs and Mann–Whitney tests, respectively. g Same as f but in the isoflurane anesthetized state (abscissa scaled to match f). *p < 0.05; **p < 0.01. Means±SE. Source data and statistical details provided in Source Data file.