Fig. 5
From: Regulation of striatal cells and goal-directed behavior by cerebellar outputs
Impact of DCN silencing on ChAT interneuron activity in the striatum. a Schematic for overview images from dorsal striatum in horizontal sections. The square indicates the location of the image in (b). Anatomical annotations: CT cortex, ST striatum, ILN intralaminar thalamic nuclei, Hippo hippocampus. b Example overview image containing parts of dorsomedial and dorsolateral striatum from CNO-injected control mice and mice expressing hM4Di in the DCN of both hemispheres. ChAT immunoreactivity in green, p-S6rp immunoreactivity in red. Scale bar: 200 µm. c, d Examples of dorsomedial striatal ChAT interneurons stained with anti-ChAT and anti-p-S6rp antibodies. Medial and interposed DCN of vGluT2-cre mice were bilaterally injected at P18 with AAV2-DiO-hM4Di and mice were intra-peritoneal (i.p.) injected with CNO (2.5 mg/kg) 50 min before analysis. Immune reactivity was examined at P35 in the dorsomedial striatum DCNctrl (control) and DCNhM4Di mouse brains. A 16-pseudocolor palette (Lookup Table) highlights the intensity of p-S6rp signal. Scale bar: 10 µm. e Cumulative frequency of p-S6rp fluorescence intensity in dorsomedial striatal ChAT neurons (n = 242 ChAT-positive neurons from N = 3 DCNhM4Di mice and from N = 3 control mice, p < 0.0001, Kolmogorov–Smirnov test). f p-S6rp signal intensity in dorsomedial ChAT-positive neurons in one hemisphere correlates with the percentage of hM4Di-expressing cells in the contralateral interposed deep cerebellar nucleus. Mice were bilaterally injected in the DCN. The infection rate in the DCN of each hemisphere was correlated with the p-S6rp immunereactvity in the contralateral striatum (r2 = 0.66, n = 242 ChAT-positive neurons across six hemispheres from N = 3 DCNhM4Di mice and N = 3 control mice without DCN injection). Boxes display and error bars display mean ± SEM). g, h As in (b, c), but analyzing p-S6rp immune reactivity in the dorsolateral striatum. Scale bar: 10 µm. i Cumulative frequency of p-S6rp fluorescence intensity in dorsolateral striatal ChAT-positive neurons (n = 134 ChAT-positive neurons from N = 3 DCNhM4Di mice and N = 3 control mice, p = 0.297, Kolmogorov–Smirnov test). j p-S6rp signal intensity in dorsolateral ChAT-positive neurons show no significant correlation to the percentage of hM4Di-expressing cells in the contralateral interposed deep cerebellar nucleus (r2 < 0.0001, n = 134 ChAT-positive neurons across six hemispheres of N = 3 DCNhM4Di mice and N = 3 control mice, boxes display and error bars display mean ± SEM)
