Figure 5.

Flow cytometry evaluation of hippocampal cLTP. (A). Top: Selection of the synaptosomal fraction based on size (FSC) and complexity (SSC). Synaptosomal membrane integrity was evaluated by Calcein AM staining, and GluA1/Nrx1β staining was analysed within Calcein⁺ events in the basal state and after chemical long-term potentiation (cLTP) stimulation. Bottom: Representative GluA1⁺/Nrx1β⁺ double-positive events at the basal state and after cLTP stimulation in the presence of the vehicle (distilled water), pramipexole (PPX) or L-DOPA. (B) The cLTP (%) at hippocampal synaptosomes in the presence of the vehicle (distilled water), PPX or L-DOPA at 1 and 4 weeks (w) post-inoculation (p.i.). (C) GluA1⁺/Nrx1β⁺ double-positive values at the basal state and after cLTP stimulation in the presence of the vehicle (distilled water), PPX and L-DOPA at 1 and 4 weeks p.i. GluA1⁺/Nrx1β⁺ values were normalized to the AAV-EVV vehicle condition for each time point, and all the values are represented as the mean ± SEM, and assessed using two-way ANOVA followed by a Tukey’s or Bonferroni’s post hoc test: ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 versus basal; ^@P = 0.0616 versus basal; *P < 0.05, **P < 0.01, ***P < 0.001 versus AAV-EVV; ^$P < 0.05 versus vehicle; ⁺P = 0.0639 versus vehicle; ^£P < 0.05, ^££P < 0.01 versus PPX (n = 8 for each group and time point). (D) Total glutamate and GABA content in the hippocampus at 4 weeks p.i. (n = 5 for each group). The values are represented as the mean ± SEM and assessed with a Mann-Whitney U-test: no significant differences. AAV = adeno-associated viral vector; EEV = empty vector; hα-syn = human α-synuclein.