Fig. 2: Tonic lPBN Y1R neuron activity tracks sustained pain.
From: A parabrachial hub for need-state control of enduring pain
a, Fibre photometry measurements of lPBN Y1R calcium dynamics. Inset, GCaMP6s expression. Scale bar, 200 μm. b, Average ΔF/F of Y1R neuron GCaMP6s signal after a hind paw formalin injection (dashed red line). Dark lines represent mean and lighter, shaded areas represent s.e.m. c, Mean ΔF/F of Y1R neurons (n = 9 mice; repeated measures one-way ANOVA, P < 0.001). d, Representative trace from an individual mouse receiving a formalin injection showing lPBN Y1R neuron activity, licking bouts and smoothed licking behaviour. e, Representative trace as shown in d along with Gaussian curves. f, Correlations between fit and signal for the two fitting methods (n = 9 mice; paired two-sided t-test on Fisher’s z values, P = 0.0055). g, A Cre-off approach to record calcium dynamics of non-Y1R neurons with image showing GCaMP6s expression in non-Y1R neurons. Scale bar, 200 μm. h, Average ΔF/F of GCaMP6s signal from non-Y1R neurons after formalin injection. i, Mean ΔF/F of lPBN non-Y1R neurons (n = 6 mice; one-way ANOVA, not significant). j, Representative trace from an individual mouse receiving formalin injection showing non-Y1R neuron activity, licking bouts and smoothed licking behaviour. k, Representative trace as shown in j along with Gaussian curves. l, Correlations between fit and signal for Y1R neurons and non-Y1R neurons (n = 9 Y1R, n = 6 non-Y1R mice; unpaired two-sided t-test on Fisher’s z values, P = 0.0006). m, A head-mounted miniature microscope was used to record activity of individual lPBN Y1R neurons. Inset, GCaMP6s expression in Y1R neurons. Scale bar, 200 μm. Units were identified using constrained nonnegative matrix factorization (CNMF)45 after video preprocessing and motion correction. n, Heat plot showing calcium fluorescence intensity in response to a formalin injection. Neuron classifications across all recorded neurons from five mice are shown. o,p, Average fluorescence (o) and examples traces (p) of neurons classified as lick-activated or pain-activated from the data shown in n. Data are mean ± s.e.m. Grey dots and lines represent individual mice.
