Fig. 4: PSMC5 overexpression alters neuronal morphology. | Nature Communications

Fig. 4: PSMC5 overexpression alters neuronal morphology.

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

Fig. 4: PSMC5 overexpression alters neuronal morphology.The alternative text for this image may have been generated using AI.

a Representative images of primary hippocampal neurons transfected with empty vector (EV) control, wild-type PSMC5 (PSMC5-WT), or PSMC5 variants. Transfected cells are labeled in red (tdTomato); MAP2 staining (blue) marks neuronal morphology. b Quantification of total neurite length shows: (i) a significant increase with PSMC5-WT (1.29 ± 0.08) compared to EV (1 ± 0.04; one-way ANOVA, F = 3.296, p = 0.0014; EV vs. PSMC5-WT, p = 0.0046, Dunnett’s multiple comparisons test); (ii) Comparable effects for variants p.(Arg201Trp), p.(Pro320His), and p.(Arg325Trp) vs. PSMC5-WT: 1.18 ± 0.10, p = 0.8; 1.19 ± 0.09, p = 0.8; and 1.14 ± 0.09, p = 0.5; respectively; and (iii) a significant decrease for variants p.(Ala202Val), p.(Thr207Met), p.(Pro320Arg), and p.(Ala324Lys360del) compared to PSMC5-WT: 0.93 ± 0.07, p = 0.0031; 1.02 ± 0.06, p = 0.02; 1.19 ± 0.09; p = 0.006; and 1.00 ± 0.07, p = 0.03, respectively, with effects indistinguishable from EV (p = 0.4, p = 0.9, p = 0.9, and p = 0.9, respectively). c Sholl analysis revealed no significant differences in dendritic arborization across conditions (one-way ANOVA, F = 2.179, p = 0.03; EV vs. PSMC5-WT: 1.12 ± 0.09, p = 0.9; PSMC5-WT vs. p.(Arg201Trp): 1.16 ± 0.14, p = 0.9; PSMC5-WT vs. p.(Ala202Val): 1.00 ± 0.12, p = 0.9; PSMC5-WT vs. p.(Thr207Met), 0.94 ± 0.06, p = 0.6; PSMC5-WT vs. p.(Pro320His), 1.36 ± 0.14, p = 0.3; PSMC5-WT vs. p.(Pro320Arg), 0.94 ± 0.06, p = 0.9; PSMC5-WT vs. p.(Arg 325Trp), 1.13 ± 0.09, p = 0.9; PSMC5-WT vs. p.(Ala324Lys360del), 1.03 ± 0.07 p = 0.9; Dunnet’s multiple comparison test). Error bars indicate SEM; n (number of neurons traced): EV = 40, PSMC5-WT = 30, p.(Arg201Trp) = 18 p.(Ala202Val) = 20, p.(Thr207Met) = 27, p.(Pro320Arg) = 29, p.(Pro320Arg) = 19, p.(Arg325Trp) = 20 and p.(Ala324Lys360del) = 20. *p < 0.05; **p < 0.01. Scale bar: 100 µm.

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