Figure 6.
Overexpression of REST/NRSF downregulates VGNa channels and downscales the global firing activity of neuronal networks. (A) Representative microphotographs of hippocampal neurons (25 div) co-infected with GFP together with either an empty vector (GFP; upper panels) or GFP+mycREST (lower panels). From left to right: GFP fluorescence (green), myc immunoreactivity (red), and DAPI staining (blue). Scale bar: 100 μm. (B) Western blot analysis of neurons expressing either GFP or GFP+mycREST and stained with anti-Nav and β-III tubulin. The bar histogram represents the mean (±s.e.m.) normalized optical density for GFP- and GFP+mycREST-expressing neurons (*P<0.05, unpaired two-tailed Student’s t-test; n=6 from three independent culture preparations). (C, D) Representative voltage traces (C) and raster plots (D) of network activity for GFP- (black) and GFP+mycREST-expressing neurons (red) at 25 div. Small vertical bars in (D) represent single action potentials. (E–G) Spiking rate (E), bursting rate (F), and burst duration (G) measured in GFP- (black symbols; n=14) and GFP+mycREST- (red symbols; n=13) expressing neurons as a function of time in vitro. Data (means±s.e.m.) were normalized to the respective values at 12 div. In GFP+mycREST-expressing neurons, a significant decrease in the spiking rate and in the bursting rate was observed (*P<0.05, **P<0.01, ***P<0.001, two-way ANOVA for repeated measurements followed by the Bonferroni’s multiple comparisons test).
Source data for this figure is available on the online supplementary information page.
