Fig 2: Selective deletion of vHPC A2ARs increases vGluT1 expression. Caffeine markedly induced vGluT1 expression in mice transfected with pAAV-EGFP but not in mice transfected with pAAV-CRE. pAAV-CRE transfection increased vGluT1 levels in mice exposed to water alone as well. Representative images (left) and Data (right) are presented as the mean ± SEM; n = 27 fields per group (3 fields per section, three sections per mouse, three mice per group). Scale bar = 30 µm.

Fig 3: Reduced density of VGluT1, VGluT2, and Calbindin in the shPMs-injected mice.(A–C) Representative immunofluorescence images showed the density of VGluT1 (A), VGluT2 (B), or Calbindin (C) in the control and indicated shPMs-injected mice. VGluT1, VGluT2, and Calbindin are markers for PF terminals, climbing fiber terminals, and PCs, respectively. (D) Quantification of the indicated markers obtained from A–C. All the data of this figure can be found in the S1 Data file. Data are shown as means ± SEMs, n = 3. Scale bar, 40 um. *P < 0.05 and ***P < 0.001. GCL, granule cell layer; ML, molecular layer; PCL, Purkinje cell layer; PF, parallel fiber; VGluT1, vesicular glutamate transporter 1; VGluT2, vesicular glutamate transporter 2.

Fig 4: Characterization of in vitro isolated SVs. (A) SVs enriched by ultracentrifugation, negatively stained by uranyl acetate. White arrows indicate the aggregation of vesicles. Scale bar = 200 nm. (B) SVs collected through home-made gel filtration column with Sephacryl S-1000 beads and negatively stained by uranyl acetate. Scale bar = 200 nm. Zoomed bar = 50 nm. (C) SVs isolated with iodixanol layer, negatively stained by uranyl acetate. Scale bar = 200 nm. Zoomed bar = 50 nm. (D) Protein levels of SV2C, vGlut1 and Rab3A were determined via western blotting. (E) SVs labeled with 10-nm gold tracer, negatively stained by uranyl acetate. Scale bar = 50 nm. (F) The measurement of the diameter of the purified SVs.

Fig 5: Gypenoside XVII prevented synaptic pruning in the prefrontal cortex (n = 4). Presynaptic protein vGluT1 in microglia (A,B). Postsynaptic protein PSD95 in microglia (C,D). ## p < 0.01 versus the Normal-vehicle group. * p < 0.05 and ** p < 0.01versus the CUMS-vehicle group.