The Journal of Neuroscience, March 7, 2007, ():
Interaction of Genetic and Environmental Factors in a Drosophila Parkinsonism Model
J. Neurosci. Chaudhuri et al.
27: 2457
Supplemental Data
Files in this Data Supplement:
- supplemental material
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Supplementary Video 1. Mobility of control (right) vs. paraquat-treated (left) wild type adults in a negative geotaxis assay. Flies fed 20 mM paraquat in 5% sucrose for 12 h have climbing deficits when compared to sibling controls fed on sucrose alone. Also, note that several of the individuals remaining near the bottom of the vial exhibit postural instability in the paraquat-treated sample.
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Supplementary Video 2. Paraquat induces parkinsonian-like bradykinesis and tremors. This higher magnification view of control (left) and paraquat-treated (20 mM, 12 h) (right) adults shows details of slowed movement. Numerous individuals exhibit body tremors.
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Supplementary Figure 1. Exposure to 20mM paraquat causes an elevation in the specific activity of catalase at 12 and 24h exposure. Specific activity is measured as units per mg protein from crude extracts of heads. One unit of catalase activity was defined as 1 mmole of H2O2 decomposed per minute. Specific activities are the averages of 6 replications derived from independently prepared extracts. Error bars indicate the standard error of the mean. ** P<0.01; * P<0.05.
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Supplementary Figure 2. Dose-dependent effects on survival during paraquat ingestion. Feeding of adult flies was initiated 24 h post-eclosion, and data were collected every 12 h until all flies had died. Greater than 94% of control flies maintained on sucrose alone were alive for the entire period of the experiment. Average survival is strongly dependent upon paraquat dosage. Vertical bars represent standard error of the mean. Each data point represents 5 replications of 15 flies each.
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Supplementary Figure 3. Effect of 20 mM paraquat (12 h exposure) on posterior dopaminergic clusters immuno-stained with anti-Drosophila TH antibody in Df(1)w, y adult brains. (A) and (C) are the control and paraquat-treated brains, respectively. The arrow in (A) indicates the PPM1 cluster which sustains neuronal loss as indicated by the arrowhead in (C). White boxes indicate the DA neuron cluster PPL1, which is enlarged in panels (B, D). The PPL1 cluster exhibits a loss of cells and rounding of remaining neurons in PQ-treated brains. Examples are indicated by arrowheads in (D), compared to the asymmetric normal neurons indicated by arrows in (B), as observed in GFP-reporter brains shown in Fig. 3. Scale bars: (A, C) 100 microns. (B, D) 20 microns.
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Supplementary Figure 4. Dopaminergic neurons are not affected by the ingestion of 1% hydrogen peroxide. (A) TH-Gal4; UAS-GFP flies at 24 h post-eclosion were fed 1% H2O2 for 60 h, an exposure time that coincided with the point of 50% mortality. There were no significant differences in dopaminergic neurons in any clusters between control and paraquat-fed flies. (B) An example of dopaminergic neurons (PPL1) after 60 h of hydrogen peroxide ingestion. Note that these neurons retain their asymmetric morphology with well-extended processes. Scale bar is 50 microns.
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Supplementary Figure 5. Catsup mutant and wild type control flies have indistinguishable catalase activity. The normal specific activity of catalase in the Catsup mutant suggests that the resistance of Catsup mutants to paraquat does not arise as a consequence of mutation-derived effects on this oxidative scavenger. Similarly, the results suggest that the sensitivity of Catsup mutants to hydrogen peroxide exposure does not appear to arise from abnormally low catalase activity. Vertical bars represent standard error of the mean.
