Figure 7.
Preservation of dopaminergic neurons in mice expressing uncleavable α-synuclein A53TN103A. (a) Immunostaining of FL α-synuclein and the N103 fragment in the SN in mice. The aggregation of α-synuclein was detected after the slides were preincubated with 10 µg/ml proteinase K. Black scale bar, 200 µm. White scale bars, 50 µm. (b) Western blots (WB) showing the expression and truncation of α-synuclein A53T and A53TN103A in the SN and striatum. L, left; R, right. α-Tubulin, loading control. Bar graph, quantification; data are mean ± s.e.m.; n = 3; *P < 0.05 by one-way ANOVA. Blots shown are representative of three independent experiments; uncropped images are shown in Supplementary Data Set 1. (c) The N103A mutation attenuates behavioral impairment induced by α-synuclein A53T. The mice underwent rotarod tests, amphetamine-induced rotation tests, cylinder tests and grid tests. Data are mean ± s.e.m.; n = 10 for the A53TN103A group; n = 9 for the GFP and A53T groups; *P < 0.05 by one-way ANOVA. (d) TH immunostaining of the striatum and SN. Right, unbiased stereological cell counts in the SN and total density of striatal dopaminergic terminals. Data are mean ± s.e.m.; n = 5; *P < 0.05 by one-way ANOVA. Scale bar, 200 µm. (e) Concentrations of dopamine in the striatal tissues, as determined by HPLC. Data are mean ± s.e.m.; n = 4; *P < 0.05 by one-way ANOVA. Source data for graphs are available online.