The interplay of seizures-induced axonal sprouting and transcription-dependent Bdnf repositioning in the model of temporal lobe epilepsy
Fig 5
The inhibition of HDAC1/2 is not sufficient to induce Bdnf transcription and repositioning.
(A) The graph shows the expression of Bdnf relative to Gapdh (normalized to control Bdnf level) in the hippocampal neurons incubated for 2 hours with DMSO vehicle (CTRL, blue bar) or picrotoxin, forskolin, and rolipram (cLTP, orange bar), incubated for 2 hours with 250 nM romidepsin and 2 hours with DMSO (romidepsin, light blue bar) or picrotoxin, forskolin, and rolipram (romidepsin+cLTP, violet bar). Kruskal-Wallis test for group comparison: p<0.01, One-Sample t-test or Welch two sample t-test for pairwise comparison: * p<0.05, *** p<0.001; error bars indicate standard error of the mean for 3 independent experiments (B) Representative picture of the nuclei of hippocampal neurons treated as described above. Hoechst 3342 staining for chromatin is shown in greyscale and segmentation of FISH signals for the Bdnf gene are shown in magenta and cyan. (C) Percentages of nuclei with the minimum distance between the respective alleles and nucleus surface < 350 nm are shown (Chi-square test, all groups, p<0.0001; Fisher’s exact tests: ** p<0.01, *** p<0.001, error bars indicate standard deviation of the binomial distribution). (D) Quantitative analysis of the intracellular positions of Bdnf alleles in the nuclei of hippocampal neurons treated and color-coded as in A. The minimal distance between the respective alleles and nucleus surface is presented in the normalized histogram.