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Fig. 2 | Acta Neuropathologica Communications

Fig. 2

From: α-Synuclein fibril-induced paradoxical structural and functional defects in hippocampal neurons

Fig. 2

Formation of α-syn inclusions in primary hippocampal neurons reduces density and head diameter of mushroom-shaped dendritic spines. Primary hippocampal neurons from wild type mice or α-syn knockout mice were exposed to fibrils or PBS at DIV 7. On DIV 12, neurons were transfected with LifeAct-GFP. Two days later (DIV 14, 7 days after adding fibrils), neurons were fixed. Widefield microscopy was used to capture Z-stacks of spines, followed by image deconvolution. a Representative images of dendritic spines in control neurons (top left), neurons 7 days after fibril induction of inclusion formation (top right). The bottom 2 panels are representative images from neurons lacking endogenous α-syn without (bottom left) or with exposure to fibrils (bottom right). Scale bars equal 5 μm. b The number of spines per 10 μm dendritic length was quantified in wild type, control neurons (blue bar; N = 7), wild type neurons after exposure to fibrils for 7 days (red bar; N = 7), α-syn knockout, control neurons (green bar; N = 6) and α-syn knockout neurons exposed to fibrils for 7 days (purple bar; N = 6). Data represents the means +/− SEM from two independent coverslips. F = 23.5, p < .0001 by ANOVA with Dunnett’s post-hoc test. c The number of thin, stubby, mushroom or filopodia shaped spines were quantified per 10 μm dendritic length. Two way ANOVA F = 5.85 (interaction), F = 17.41 (treatment) ** represents p < .0001. d Cumulative frequency plot of spine extent (length). P = 0.95, Kruskal-Wallis test. e Cumulative frequency plot of spine head diameter. P = 0.001 Kruskal-Wallis test. f Spine head diameter of thin, stubby, mushroom or filopodia shaped spines was quantified for wild type control neurons and neurons with α-syn inclusions. *** represents p < .0001 by independent t-test

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