Fig. 1: Ethanol impairs vigilance-dependent cerebellar BG Ca2+ activation.
a Scheme of 2P Ca2+ imaging in awake, head-restrained mouse on a motorized linear treadmill. b Left, Pseudocoloured in vivo Ca2+ image of BG processes, tangential optical section—cerebellum molecular layer, Slc1a3-CreERT;R26-lsl-GCaMP3 mouse. Right, Locations of regions of interest (ROIs) used in c. c Upper, Pseudocolour plot of all ROIs’ Ca2+ responses. Green bars, enforced locomotion. Lower, Corresponding average Ca2+ response trace of all ROIs (dark red) and traces representing numbered ROIs. d Time course of effect of saline/ethanol injection (red dotted line) on ΔF/F10s (normalized mean Ca2+ change ((F − Fmedian of baseline)/Fmedian of baseline7) within 10 s from onset of locomotion). Normalized to average first five trials (baseline). Blue bar, effect analysis time window. mean ± SEM, n = 6 mice per dosage. e Upper, Maximum response plots during indicated experimental episodes. Scale bar, 100 μm. Lower, Linear Pearson correlation coefficient plots between individual ROIs’ Ca2+ change traces before and after 2 g/kg ethanol. Numbers of independent experimental repetitions with similar results were: 6 (saline), 7 (0.5 g/kg), 6 (1.0 g/kg), 6 (1.5 g/kg), and 6 (2.0 g/kg). f–i Population data, mean normalized values within blue bar in d. Numbers under abscissa, g/kg i.p. ethanol. Data represent: mean ΔF/F10s (f), mean time to peak (baseline time from onset of locomotion to peak of population response; g), mean correlation coefficient (baseline Pearson linear correlation coefficient among Ca2+ change traces within 20 s from onset of locomotion (r); h) and coefficient of variation (CV) among ΔF/F traces of individual ROIs (i). Red symbols, mean ± SEM if data follow Gaussian distribution; without error bars, red symbols represent median if data do not follow Gaussian distribution. Lines between dots support readability. n = 6 mice per dosage; one-way ANOVA (f F(5, 30) = 11.110, p < 0.001; g F(5, 30) = 4.963, p = 0.002 and i F(5, 30) = 9.543, p < 0.001) or Kruskal–Wallis test (h) were followed by Tukey–Kramer correction; n.s. not significant. Source data are provided as a Source Data file.
