Comparison between groups Ganetespib research buy in bDNA assays was carried out using Student’s t-test after checking the normal distribution of values with Normal QQ plots test and the variance within groups with the F-test. It is of note that for SV2C values, the MTS1A group showed a much higher variance than the other groups, did not approximate a Gaussian distribution and rather assumed a bimodal pattern (see Figure 1). The analysis of covariation between dynorphin, ZnT3 and SV2C IR scores and SV2C mRNA levels was performed using the Spearman rank correlation test. Correlation was considered significant for two-tailed P-value < 0.05. Statistical analysis was carried out using the

R-cran statistical software (R Core Team [34]) (Table 3). Quantitative mRNA data on the expression of SV2A, SV2B and SV2C are shown in Figure 1 and individual

values are given in supplementary Table S1. Experiments have been carried out in triplicate and graphs show the mean value of the three experiments. SV2A and SV2B expression was globally decreased in cases of MTS and gliosis, reflecting the overall synaptic loss. All comparisons between TLE groups and controls reach statistical significance with P-values ≤ 0.05 Palbociclib solubility dmso (see Figure 1). SV2C mRNA was globally increased in the group of MTS1A, and this increase was statistically significant using Student’s t-test. The MTS1A group appeared heterogeneous however, with five cases showing high levels of SV2C (NC1, NC6, NC26, NC28 and NC33) while the other 13 showed mild or no SV2C increase. There was an excellent agreement between mRNA quantification and IR data indicating that overexpression of SV2C occurs at both mRNA and protein level (see text below and Table 3). The five cases that were positive by both methods have been labelled with colours in Figure 1. We used immunohistochemistry to identify the distribution pattern of

SV2 isoforms in controls and TLE cases. In autopsy controls, SV2A and SV2B IR was seen in all subfields of the hippocampus and closely matched the pattern of synaptophysin, as expected for a selective presynaptic staining (Figure 2b–d) [19, 35]. It is of note that SV2B IR was consistently weaker than SV2A mafosfamide and synaptophysin, particularly in the CA4 and CA3 areas. Most often no staining for SV2C was seen throughout the hippocampus (Figures 2e and 3a). Only in rare cases, there was a faint staining confined to synapse aggregates in CA4. These results suggest that, while SV2A and SV2B are widely distributed, SV2c expression, when present, is restricted to the axonal projections of neurones from the granular cell layer of the dentate gyrus (GCL), the so-called mossy fibre pathway targeting CA3 and CA4 pyramidal neurones [36, 37]. We compared SV2C with dynorphin IR, as this opioid peptide is expressed in mossy fibres [22]. As expected, in controls dynorphin IR was seen in GCL, in the innermost portion of the molecular layer, in the hilus (CA4) and in the stratum lucidum (CA3) (Figure 3b).