Fig. 3: Psmc5 suppression impairs reversal learning in flies and alters synaptic balance in rat hippocampal neurons. | Nature Communications

Fig. 3: Psmc5 suppression impairs reversal learning in flies and alters synaptic balance in rat hippocampal neurons.

From: Investigating the neuronal role of the proteasomal ATPase subunit gene PSMC5 in neurodevelopmental proteasomopathies

Fig. 3: Psmc5 suppression impairs reversal learning in flies and alters synaptic balance in rat hippocampal neurons.The alternative text for this image may have been generated using AI.

All data are presented as mean±SEM. a–c Pan-neuronal KD of Rpt6 (PSMC5) in Drosophila had no effect on normal olfactory learning (two-sided Student’s t-test; wild-type WT: 78 ± 2.9; KD(Elav:Rpt634712): 82 ± 1.7; p = 0.3059; n = 4 biological replicates of PI), but significantly impaired reversal learning (one-way ANOVA with Tukey post hoc; WT: 43 ± 1.9; WT:Elav: 40 ± 1.7; WT:Rpt634712: 42 ± 3; KD(Elav:Rpt634712): 1.5 ± 2; p < 0.0001, n = 4 PI), with age-persistent deficits. ***p < 0.001; ****p < 0.0001. d–j Climbing activity declined in Rpt6 KD flies over time: at 1–3 days (two-sided t-test; WT: 81.5% ± 5.8%; KD: 42.9% ± 4.4%; p < 0.0001; n = 10 biological replicates); at 6–8 days (two-sided t-test; WT: 81.5% ± 5.5%; KD: 43.1% ± 9.1%; p = 0.0021; n = 10 biological replicates); at 22–24 days (t two-sided t-test: 82.7% ± 2.7%; KD: 12.7%±2.5%; p < 0.0001; n = 10 biological replicates). WT flies maintained performance across all ages. **p < 0.01; ***p < 0.001. k–n Primary rat hippocampal neurons infected with lentiviral particles (GFP + scrambled or Psmc5-KD shRNA) showed: reduced PSMC5 protein in GFP positive (+) Psmc5 KD neurons compared to scrambled controls (two-sided Mann Whitney test; control: 100% ± 36.76%; KD: 12.71% ± 1.61%; p = 0.0002, n = 10 images analyzed per condition); and increased ubiquitin accumulation (two-sided Mann Whitney test; control: 2.8 × 106 ± 1.3 × 105; KD: 4.8 × 106 ± 5.6 × 105; p = 0.0022, n = 6). Scale bar = 20 μm. o, p No significant difference in dendritic branching was observed at DIV14 between Psmc5 KD and control neurons (two-sided Mann-Whitney test). The two groups exhibited comparable numbers of primary dendrites (control: 3.67 ± 0.33; Psmc5 KD: 3.83 ± 0.31; p = 0.749), secondary dendrites (control: 13.00 ± 1.37; KD: 13.17 ± 2.10; p = 0.873) and tertiary dendrites (control: 17.50 ± 6.69; KD: 18.33 ± 1.73; p = 0.575). n = 6 neurons per condition. MAP2 staining of infected neurons is shown in red. Scale bar: 20 μm. q, r Synaptic analysis revealed: Reduced vGlut⁺ excitatory puncta (two-sided Mann Whitney test; control: 1.2 ± 0.1; KD: 0.8 ± 0.1; p = 0.0045; n = 5) and stable vGat⁺ (two-sided Mann Whitney test; control: 0.8 ± 0.08; KD: 1.0 ± 0.2; p = 0.2988; n = 5) in KD neurons. s, t Decreased vGlut⁺ (filled arrows) and stable vGat⁺ (open arrows) signals in KD vs. control neurons. *p < 0.05; **p < 0.01. Scale bar = 5 μm.

Back to article page