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. 2018 Feb 20;11:45. doi: 10.3389/fnmol.2018.00045

Figure 6.

Figure 6

Administration of a caspase-3 inhibitor before P7 ethanol treatment prevents long-term potentiation (LTP) deficits in adult male mice. (A) A schematic drawing shows the stimulating and recording electrode positions in the CA1 region of the HP. (B) The average field-excitatory-post-synaptic potential (fEPSP) slope at various time points obtained from P7 saline (S), ethanol (E), S + Q-VD-OPh, or E + Q-VD-OPh-treated adult mice. For each slice, the fEPSP slopes were normalized against the average slope over the 10-min recording before LTP stimulation. Arrows show the time of Theta-Burst Stimulation (TBS; four pulses at 100 Hz, with bursts repeated at 5 Hz, and each tetanus including three different 10-burst trains separated by 15 s). Inset: representative traces before and after TBS are shown for all the treatment groups. Scale 1 mV; 10 ms. (C) Bar graph compares the average of the fEPSP slopes at several time points after TBS in P7 saline (S), ethanol (E), S + Q-VD-OPh, or E + Q-VD-OPh-treated adult mice hippocampal slices. (*p < 0.01 vs. saline, n = 5 mice/group; 10 slices/group). (D) Scheme indicating the experimental design to evaluate the effects of pre-administration of Q-VD-OPh before saline/ethanol treatment in P7 mice on activity-dependent signaling events in adulthood. The spatial working memory of adult mice treated with saline (S), ethanol (E), S + Q-VD-OPh, or E + Q-VD-OPh at P7 was evaluated using a Y-maze. (E) HP tissues were collected immediately after a 24 h intertrial interval when the mice completed exploring all three arms (3 min, preference trial, test trial). The signaling proteins (E1) pCaMKIV, (E2) pCREB, (E3) pMeCP2 and (E4) Arc were examined in nuclear or cytosolic extracts of the HP from saline (S), ethanol (E), S + Q-VD-OPh, or E + Q-VD-OPh groups (n = 8 mice/group) via western blot analysis. Equal protein loading was confirmed after Ponceau S staining, and β-actin was used as a protein loading control (*p < 0.01 vs. S; #p < 0.01 vs. E; &p < 0.01 vs. S+V; $p < 0.01 vs S; @p < 0.01 vs E. Error bars, SEM (two-way ANOVA with Bonferroni’s post hoc test).