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Fig. 2 | Acta Neuropathologica Communications

Fig. 2

From: Progressive striatonigral degeneration in a transgenic mouse model of multiple system atrophy: translational implications for interventional therapies

Fig. 2

Biochemical analysis of α-synuclein expression in MSA transgenic mice during aging. a ELISA measurement of α-syn in whole brains shows 5 to 8 times higher α-syn levels in MSA than in control mice, with no significant changes over time in either group. b Subcellular fractionation of hemi-brains and subsequent Western immunoblotting using either the C20 polyclonal or the Syn1 monoconal Ab (n = 4 in each group) reveals higher α-syn levels in the TX-soluble fraction in MSA vs. control mice (two-way ANOVA with factors genotype and age: Syn1: genotype F1,24 = 12.58, p < 0.0016, age F3,24 = 1.244, p > 0.05; C20: genotype F1,24 = 14.38, p < 0.0009, age F3,24 = 1.505, p > 0.05); however, sub-group (age) differences are not significant following post-hoc Bonferroni test. Oligomeric species expression, assessed with the Syn1 Ab, tended to increase from 6 months onwards (Syn1: genotype F1,24 = 11.36, p < 0.0026, age F3,24 = 1.908, p > 0.05) without significant sub-group differences. SDS-soluble fractions reveal consistent increased monomeric α-syn levels regardless of age (C20: 2, 6, 12, 18 months: p < 0.001; 0.001; 0.05; 0.001) and HMW oligomeric species expression (again assessed with the Syn1 Ab) that peaked at 12 months (p < 0.001). The asterisk denotes a non-specific band at 25 kDA that is not included in the quantification of HMW species. c Subregional analysis by ELISA of MSA mouse brains (n = 4–5 for all groups) reveals an increase in α-syn levels with time only in the midbrain, from 12 to 18 months of age; the lowest α-syn levels are seen in the forebrain, while the highest appear in the lower brainstem (two-way ANOVA with factors region and age: effect of region F3,56 = 53.86, p < 0.0001, effect of age F3,56 = 1.104, p = 0.3552, interaction F9,56 = 2.042, p = 0.0512). (+ p < 0.05, ++ p < 0.01, +++ p < 0.001, * p < 0.05). On the side – representation of the dissection protocol of the subregions. d Western blot analysis confirms significantly higher α-syn expression in the MSA than in the control wildtype mice in all the considered areas at 2 and 12 months of age (n = 6 for each group)

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