Analysis of inflammation-related nigral degeneration and locomotor function in DJ-1 −/− mice
- Equal contributors
1 Department of Physiology, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX 75390, USA
2 Department of Neurology and Neurotherapeutics, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX 75390, USA
3 Department of Psychiatry, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX 75390, USA
4 Department of Physiology, Emory University School of Medicine, 615 Michael Street, Atlanta, GA, 30322, USA
Journal of Neuroinflammation 2013, 10:50 doi:10.1186/1742-2094-10-50Published: 28 April 2013
Additional file 1: Figure S1:
Schematic of intranasal TNF (inTNF) dosing paradigm in DJ-1−/− and wild type mice and effects on nigral DA neuron number and striatal DA. A, Schematic of experimental design. Wild type and DJ-1−/− mice were administered soluble murine TNF intranasally at the indicated concentrations or an equivalent volume of saline vehicle for the indicated times. Groups of mice were sacrificed as indicated for biochemical (striatal DA measurements by HPLC) and immunohistological (unbiased stereological estimate of nigral DA neuron number) analyses. B, Unbiased stereological analysis indicates that DJ-1−/− mice exposed to repeated doses of 0.5 ng TNF intranasally do not display a significant reduction of TH or NeuN immunopositive neurons in the SNpc. Error bars represent SEM, n=6-10 per group. Two-way ANOVA indicated no significant differences (TH: p = 0.085, NeuN: p = 0.257). C, Striatal dopamine (DA) was measured by HPLC with electrochemical detection and striatal DA turnover was calculated as the ratio of DA metabolites (DOPAC, HVA and 3-MT) to DA. Error bars represent SEM, n = 6–10 per group. Two-way ANOVA, Bonferroni’s post hoc **p < 0.01 compared to wild type saline (DA: F(1,29)= 22.07, p < 0.0001, DA turnover: not significant, p = 0.055).
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Additional file 2: Figure S2:
Striatal levels of DA and metabolites for wild type and DJ-1−/− mice in the 3-month treatment group was measured by HPLC electrochemical detection. (DOPAC, HVA, and 3-MT were not significant by two-way ANOVA, p > 0.05 for all).
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Additional file 3: Figure S3:
Neuroinflammatory and oxidative stress responses of isolated peritoneal macrophages from DJ-1−/− mice are similar to wild type. Primary peritoneal macrophages were isolated from wild type and DJ-1−/− mice and treated with 1 ug/mL LPS for 4 hrs. QPCR was performed for gene expression of TNF, iNOS, IL1b, NRF2, and NQO1. A two-way ANOVA was performed with Bonferroni’s post hoc at * for p < 0.05, ** for p < 0.01, and *** for p < 0.001 significance compared to the saline treatment. No genotype differences were detected (TNF: p = 0.95, iNOS: p = 0.33, IL1β: p = 0.60, NRF2: p = 0.54, NQO1: p = 0.09).
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