Article Figures & Data
- Figure 1
Generation of DJ-1 knockout Polg mutator mice. A, Double heterozygous mice were bred to generate the four genotypes of mice used in this study. B, The number of viable pups born from double heterozygous breeding is shown. A χ2 test was used to determine that the observed proportions did not differ from the expected proportions (n = 208 mice, p = 0.901)a. C, The weights of female mice at one year of age are displayed. The groups were compared with ANOVA (F(3,9) = 29.17, p = 0.00005745)b followed by Tukey’s multiple comparison test (*p < 0.05 vs DJ-1+/+;PolgWT/WT , $p < 0.05 vs DJ-1−/−;PolgWT/WT ). D, The weights of male mice at 1 year of age.
- Figure 2
Behavioral characterization using the pole test. A, The mice were tested for behavioral deficits using the pole test and the method of descent for each mouse during each of their trials is displayed. Each bar represents an individual animal, and the methods of descent from four to seven trials are reported as a proportion within the bar. B, The mean time to descend the pole for each mouse is displayed (n = 4-7 mice per genotype, n = 4-7 trials per mouse, ANOVA F(3,19) = 1.171, p = 0.347)c. C, Following their first set of trials on the pole test, three mice in the DJ-1+/+;PolgMT/MT group and three mice in the DJ-1−/−;PolgMT/MT were given l-DOPA and retested 30 min later. The results of the test before and after l -DOPA are displayed with each bar representing an individual animal (n = 4-7 trials).
- Figure 3
Stereological counts of dopaminergic neurons in the SNpc. Unbiased stereology was performed by a blinded observer to count the number of dopaminergic neurons in the SNpc of the mice after that had reached a year of age. Two separate experiments were performed to analyze the same set of brains. A, TH immunoreactive cells in the midbrain stained brown using DAB (scale bars, 200 μm). B, DAB-stained cell counts for each animal (red circles) along with mean and SEM of each group (n = 6-8 mice per genotype, ANOVA F(3,23) = 2.072, p = 0.1318)d. C, TH immunoreactive cells were detected in the midbrain using fluorescence (TH = green; scale bars, 500 μm). D, The numbers of SNpc dopaminergic neurons counted using stereology for each animal (red circles) are shown with mean and SEM (n = 6-8 mice per group, ANOVA F(3,23) = 0.9124, p = 0.4504)e.
- Figure 4
Dopaminergic terminal density in the striatum. A, Representative TH-stained tissue sections through the striatum. The sections were immunostained using an infrared fluorescent dye conjugated secondary antibody and imaged using an infrared imaging system. The sections are pseudo-colored using a heat map, with warmer colors indicating strong TH immunoreactivity. B, Striatal TH staining intensity calculated from infrared imaged tissues. Individual data points represent animals and the mean and SEM are also displayed (three sections per animal were averaged, n = 5-8 animals, ANOVA F(3,22) = 1.189, p = 0.3369)f.
- Figure 5
Detection of astrogliosis in the SNpc. A, GFAP-positive astrocytes were immunostained in the SNpc (outlined in white) and surrounding tissue (GFAP = red; scale bars, 500 μm). Unbiased stereology was used to count GFAP-positive cells in the SNpc simultaneously with the TH cells counts shown in Figure 3B. B, GFAP-positive SNpc cell counts per animal (red circles) along with mean and SEM are displayed in the graph (ANOVA F(3,23) = 1.744, p = 0.1860)g.
Data structure Type of test Power (f = 0.25) a Χ2 0.999 b N too small to determine if normally distributed One-way ANOVA 0.0001 c N too small to determine if normally distributed One-way ANOVA 0.471 d N too small to determine if normally distributed for 3 of 4 genotypes. The fourth genotype with 8 animals is normally distributed. One-way ANOVA 0.274 e N too small to determine if normally distributed for 3 of 4 genotypes. The fourth genotype with 8 animals is normally distributed. One-way ANOVA 0.632 f N too small to determine if normally distributed for 3 of 4 genotypes. The fourth genotype with 8 animals is normally distributed. One-way ANOVA 0.493 g N too small to determine if normally distributed for 3 of 4 genotypes. The fourth genotype with 8 animals is normally distributed. One-way ANOVA 0.350 Post hoc power calculations for each statistical test reported are recorded in this table. To calculate post hoc power, we used an effect size of 0.25, a sample size that was the minimum group size, and the actual p value returned by the indicated test. For almost all instances, we were unable to formally test whether the data were normally distributed because the sample size was not 8 or greater. Superscript letters in the figure legends refer to the first column of the table.
DJ-1 Polg Male Female Total +/+ WT/WT 1 5 6 −/− WT/WT 3 4 7 +/+ MT/MT 3 3 6 −/− MT/MT 5 3 8 The genotype and gender of the cohort of 27 mice used for experiments are recorded in the table.
- Movie 1
Pole test of a DJ-1+/+;PolgWT/WT mouse. The mouse performed the task correctly be reorienting itself and walking down the pole.
- Movie 2
Pole test of a DJ-1−/−;PolgWT/WT mouse. This mouse performed the task correctly by walking down the pole.
- Movie 3
Pole test of a DJ-1+/+;PolgMT/MT mouse. The mouse did not perform the task correctly because it did not orient itself downwards and because it slid down the pole.
- Movie 4
Pole test of a DJ-1−/−;PolgMT/MT mouse. The mouse did not perform the task correctly because it fell from the top of the pole.
- Movie 5
Pole test of a DJ-1+/+;PolgMT/MT mouse given l-DOPA. This mouse (also shown in Movie 3) was given l -DOPA 30 min prior to the test. It did not perform the task correctly, but instead slid down the pole.
- Movie 6
Pole test of a DJ-1−/−;PolgMT/MT mouse given l -DOPA. This mouse (also shown in Movie 4) was tested 30 min after l -DOPA administration. It did not perform the task correctly, but instead fell from the top of the pole.
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