Fig. 7. Chemogenetic and optogenetic inhibition of Bar^ESR1 neurons impairs voluntary scent marking urination.

a, Schematic of chemogenetic inhibition of Bar during scent marking urination. b, Example hM4Di expression in Bar of ESR1-Cre, top, and CRH-Cre, bottom, mice; larger views minus Nissl on the right. c, Number of Bar cells infected with hM4Di virus versus CNO urine inhibition index (see Methods) for all mice (green for ESR1-Cre, magenta for CRH-Cre). d, Raster plots of urine marks on consecutive days with either CNO or saline (Bar^ESR1-hM4Di, top, Bar^CRH-hM4Di, middle, CNO-only control, bottom). e, Percentage of maximum urine marks across all CNO or saline days for, top, Bar^ESR1-hM4Di (n=8, p=0.00013 Friedman’s test, **day 2 saline p=0.0058, day 4 saline p=0.011 Dunn-Sidak posthoc differences from CNO days 1 & 3, middle, Bar^CRH-hM4Di (n=10, p=0.29 Friedman’s test) and, bottom, CNO control (n=7, p=0.86 Friedman’s test) mice (thin lines individual mice, thick lines mean ± s.e.m.). f, Schematic of optogenetic inhibition of Bar^ESR1 during scent marking urination. g, Δurine amount around 2 min. photoinhibition period. Female odor presented within 15 seconds of light on, and subsequent sniff periods shown in blue. n=9 trials from 3 mice. h, Δurine amount ± 5 sec. from end of photoinhibition for control odor and female odor (thick line and shading are mean ± s.e.m., n=9 total trials from 3 mice). i, Urine amount (**p=0.0039), and, j, female odor sniff time (p=0.20) during 2 min. photoinhibition period and 2 min. immediately following (mean ± s.e.m., same trials as h, Wilcoxon signed rank tests). Green shading denotes photoinhibition periods. Scale bars = 100 μm.