Fig. 1 Motor coordination deficits, psychostimulant hypersensitivity, and sensorimotor gating deficits in Df1/+ mice. (A) Latency to fall (in seconds) from the rotarod was scored three times per day over three consecutive days. WT control (n = 10) and Df1/+ (n = 10) mice were tested at 3.5 months of age. Statistical significance [two-way analysis of variance (ANOVA) with repeated measures: F1,18 = 14.09, P = 0.0015]; *P < 0.05 (Bonferroni post hoc test). (B) Locomotor activities [distance traveled (in centimeters)] measured in the open-field box before and after MK-801 challenge were plotted in 15-min bins. WT (n = 10) and Df1/+ (n = 10) mice were tested at 2 months of age. Statistical significance (two-way ANOVA with repeated measures: F1,18 = 4.763, P = 0.0426); **P < 0.01 (Bonferroni post hoc test). (C) Acoustic startle response (left) and PPI (%) of startle response (right) were evaluated. WT (n = 10) and Df1/+ (n = 10) mice were tested at 2 months of age. Statistical significance (two-way ANOVA with repeated measures: F1,18 = 8.528, P = 0.0091); *P < 0.05 and **P < 0.01 (Bonferroni post hoc test). a.u., arbitrary units.
Fig. 2 Elevated expression of α-synuclein and p62 proteins in Df1/+ mice. (A and B) The ACC, CPu, and SNc in Df1/+ mice (n = 4) and the WT littermates (n = 4) (3.5 months old) were immunostained with anti–α-synuclein (α-Sync) (A) or anti-p62 (B) antibodies, together with anti-CaMKII, anti–DARPP-32, and anti-TH antibodies, which label the most abundant neuronal cell type in the ACC, CPu, and SNc, respectively. Images were acquired by a confocal microscope using the same parameters across multiple specimens. Images in the ACC and SNc are shown. Scale bars, 20 μm. Relative fluorescence intensities (arbitrary units) of α-synuclein or p62 immunostaining were measured from each soma colabeled with the neuronal marker (30 to 50 soma per section) and plotted in the graph. Overall average fluorescence intensities (±SEM) calculated across three to four serial sections per animal from all animals used (n = 4 per group) are shown in red in the graph. *P < 0.05 and **P < 0.01 (two-tailed Mann-Whitney test).
Fig. 3 Genetic or pharmacological rescue of motor coordination deficits, psychostimulant hypersensitivity, and sensorimotor gating deficits in Df1/+ mice. (A) Latency to fall (in seconds) from the rotarod was scored three times per day over three consecutive days. WT control (n = 8), Df1/+ (n = 8), Df1/+;Snca/+ (n = 8), and Snca/+ (n = 8) mice were tested at 3.5 months of age. Statistical significance (two-way ANOVA with repeated measures: F3,28 = 9.552, P = 0.0250); *P < 0.05 (Df1/+, as compared to WT) and #P < 0.05 (Df1/+, as compared to Df1/+;Snca/+) (Bonferroni post hoc test). (B) Latency to fall (seconds) from the rotarod was scored three times per day over three consecutive days. WT mice treated with vehicle (n = 10), Df1/+ mice treated with vehicle (n = 12), and Df1/+ mice treated with CCI-779 (n = 8) were tested at 3.5 months of age. Statistical significance (two-way ANOVA with repeated measures: F2,27 = 4.34, P = 0.0232); *P < 0.05 (Df1/+ mice treated with vehicle, as compared to WT) and #P < 0.05 (Df1/+ mice treated with CCI-779, as compared to Df1/+ mice treated with vehicle) (Bonferroni post hoc test). (C) Locomotor activities [distance traveled (in centimeters)] measured in the open-field box before and after MK-801 challenge were plotted in 15-min bins. WT mice treated with vehicle (n = 10), WT mice treated with CCI-779 (n = 10), Df1/+ mice treated with vehicle (n = 10), and Df1/+ mice treated with CCI-779 (n = 10) were tested at 2 months of age. Statistical significance (two-way ANOVA with repeated measures: F3,36 = 3.132, P = 0.0374); *P < 0.05 and **P < 0.01 (Bonferroni post hoc test). (D) Acoustic startle response (left) and PPI (%) of startle response (right) were evaluated. WT mice treated with vehicle (n = 10), WT mice treated with CCI-779 (n = 10), Df1/+ mice treated with vehicle (n = 10), and Df1/+ mice treated with CCI-779 (n = 10) were tested at 2 months of age. Statistical significance (two-way ANOVA with repeated measures: F3,36 = 4.347, P = 0.0103); *P < 0.05 and **P < 0.01 (Bonferroni post hoc test).
- Table 1 Expression of α-synuclein and p62 proteins in Df1/+ mice.
ND, not determined; ↑, up-regulated (>1.5×, compared with WT); ↗, up-regulated (<1.5×, compared with WT); →, unchanged (compared with WT).
ACC CPu SNc α-Synuclein p62 α-Synuclein p62 α-Synuclein p62 2 months ND ↑ ND ↗ ND → 3.5 months ↑ ↑ ↑ ↑ ↑ ↗ 8 months ↑ ↑ ↗ → ↑ →
Supplementary Materials
Supplementary material for this article is available at http://advances.sciencemag.org/cgi/content/full/4/8/eaar6637/DC1
Fig. S1. Amplitude of responses to nonstartle sound stimuli in Df1/+ mice during PPI assays.
Fig. S2. Elevated expression of α-synuclein and p62 in Df1/+ mice at 8 months of age.
Fig. S3. Elevated p62 expression in Df1/+ mice is normalized by CCI-779 administration.
Fig. S4. Increased mTOR activity in Df1/+ mice is normalized by CCI-779 administration.
Additional Files
Supplementary Materials
This PDF file includes:
- Fig. S1. Amplitude of responses to nonstartle sound stimuli in Df1/+ mice during PPI assays.
- Fig. S2. Elevated expression of α-synuclein and p62 in Df1/+ mice at 8 months of age.
- Fig. S3. Elevated p62 expression in Df1/+ mice is normalized by CCI-779 administration.
- Fig. S4. Increased mTOR activity in Df1/+ mice is normalized by CCI-779 administration.
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