Fig. 2: ASIC1a regulates the hypothalamic signaling activity.

a Cultured hypothalamus neurons were classified into types 1–3 as indicated in Supplementary Fig. 2a. Neurons were exposed to current protocols, indicated in Supplementary Fig. 2c, to determine the proportion of secretory and non-secretory neurons. b–e data obtained after synchronization by dexamethasone of cultured mouse neurosecretory Type 1 hypothalamus neurons. b Peak current densities of pH6.6-induced currents at the indicated ZT, measured by voltage-clamp at −60 mV; n = 10-11 cells per condition. c–e Current-clamp measurements; c Representative voltage trace of pH6.6-induced depolarization. d Number of pH6.6-induced action potentials (APs) at the indicated ZT; n = 8–10 cells per condition. e Normalized AUC of the depolarization (voltage · time, gray area in c) of pH6.6-induced depolarization at the indicated ZT; n = 8–10 cells per condition. f–h Extracellular recordings, from mouse brain slices. f Scheme of the coronal PVH slice map, from which recordings in loose-patch mode were done on mpd neurons. pv, periventricular parvocellular; dp, dorsal parvocellular; pml, posterior magnocellular lateral; pmm, posterior magnocellular medial; III, third ventricle. g Representative traces recorded during the day or night as indicated, from WT (blue) and ASIC1a^−/− (red) brain slices. h Mpd neuron SFR from WT and ASIC1a^−/− mice. Cells not firing APs were not included in this analysis; n = 26–30 cells from four mice in each condition. Statistical analysis was done on log2-transformed data for b, e, h. One-way ANOVA test and Tukey post hoc test (b, e), Kruskal–Wallis and Dunn’s post-hoc test of non-transformed data (d); two-way ANOVA and Tukey post hoc test (h). ^#p < 0.05; ^###p < 0.001^{; ####}p < 0.0001 (b–e, h), comparison between day and night or different ZT for the same genotype. ***p < 0.01; WT compared to ASIC1a^−/− at the corresponding period (h). Error bars indicate SEM.