Fig. 7
CF activity is required for C1ql1 to induce CF innervation on adult PCs
A Experimental scheme for silencing of CFs and recording of CF-EPSC. B Representative light-evoked CF-EPSC traces (blue lines). Multiple EPSCs with a slower rise time were recorded in slices overexpressing C1ql1 in CFs (B1). The boxed region is enlarged on the right to show the slow CF-EPSC. Single EPSC was evoked in an all-or-none manner in slices overexpressing C1ql1 and ESKir2.1 in CFs (B2). C Total CF-EPSC amplitude. p = 2.665 × 10− 7, two-sided Welch’s t-test. n = 27 cells from 5 mice (CTRL), n = 31 cells from 5 mice (ESKir2.1). D The percentage of the number of CFs innervating single PCs. The number of EPSCs evoked by distinct CF activation thresholds (number of steps) is shown. p = 0.0221, Mann–Whitney U test. n = 34 cells from 5 mice (CTRL), n = 34 cells from 5 mice (ESKir2.1). E Experimental scheme to study the effect of CF neural activity on C1ql1 immunoreactivity. F Immunohistochemical analysis of HA-C1ql1 at CF synapses. Expression of ESKir2.1 (GFP, bottom panels) in IOs reduced C1ql1 (HA) immunoreactivity at CF synapses (vGluT2, arrowheads) compared to ESKir2.1AAA (GFP, top panels). Scale bar, 10 μm. G Immunohistochemical analysis of HA-C1ql1 in IONs. Scale bar, 20 μm. H Quantification of HA-C1ql1 levels at CF synapses. HA-C1ql1 immunoreactivity was normalized by the GFP fluorescence in vGluT2-positive CF synapses. p = 2.353 × 10− 4, two-tailed Welch’s t-test. n = 103 areas from 3 mice (CTRL), n = 169 areas from 3 mice (ESKir2.1). I Quantification of HA-C1ql1 levels in IONs. HA-C1ql1 immunoreactivity was normalized by the GFP fluorescence in the soma of IONs. p = 0.5970, two-tailed Welch’s t-test. n = 149 cells from 3 mice (CTRL), n = 161 cells from 2 mice (ESKir2.1). Bars represent mean ± SEM. **p < 0.01; *p < 0.05; ns, not significant