DJ-1 attenuates the glycation of mitochondrial complex I and complex III in the post-ischemic heart

Figures

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  Figure 1. Deficiency of DJ-1 Enhances ischemia–reperfusion induced mitochondrial glycative stress. Levels of (A) methylglyoxal (MG), (B) advanced glycation end products (AGE), and (C) carboxymethyllysine (CML) in mitochondrial fractions. All measurements were performed in samples collected at 3 days of reperfusion from Wild-Type and DJ-1 KO mice. Values are means ± SEM. 5 samples per group. Two-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, and ***p < 0.001 vs. Wild-Type Sham.

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  Figure 2. Ischemia–reperfusion injury enhances the localization of DJ-1 to the mitochondria. (A) Immunoblots and (B) analysis of the full-length (DJ-1FL) and cleaved (DJ-1ΔC) forms of DJ-1 in mitochondrial fractions. All measurements were performed in samples collected at 3 days of reperfusion from Wild-Type mice. Values are means ± SEM. 6 samples per group. Student’s T-test. **p < 0.01 vs. Sham.

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  Figure 3. Ischemia–reperfusion injury enhances the interaction of DJ-1 with complex I and complex III of the mitochondrial electron transport chain. (A) Immunoblots and (B) analysis of the interaction between DJ-1 and complex I of the mitochondria electron transport chain. (C) Immunoblots and (D) analysis of the interaction between DJ-1 and complex III of the mitochondria electron transport chain. Measurements were performed in samples collected at 3 days of reperfusion from Wild-Type mice. Values are means ± SEM. 4–6 samples per group. Student’s t test. **p < 0.01 vs. Sham.

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  Figure 4. Overexpression of Cleaved form of DJ-1 attenuates ischemia–reperfusion induced glycation of complex I and complex III. (A) Immunoblots and (B) analysis from immunoprecipitation experiments examining the glycative status of complex I of the mitochondria electron transport chain. (C) Complex I activity. (D) Immunoblots and (E) analysis from immunoprecipitation experiments examining the glycative status of complex III of the mitochondria electron transport chain. (F) Complex III activity. Measurements were performed in samples collected at 3 days of reperfusion from wild-type and DJ-1 KO mice administered AAV9-GFP (GFP) or AAV9-DJ1Δc (DJ1Δc). Values are means ± SEM. 4–6 samples per group. Two-way ANOVA with a Tukey test as the posthoc analysis. ***p < 0.001 vs. wild-type Sham. IP immunoprecipitation. IB immunoblot.

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  Figure 5. Overexpression of cleaved form of DJ-1 attenuates ischemia–reperfusion induced mitochondrial dysfunction. (A) Basal, (B) maximum (State 3), and (C) oligomycin (Oligo) oxygen consumption rates in the presence of pyruvate and malate. (D) Respiratory control ratio (RCR; State 3/oligomycin). (E) ATP production rates. (F) Efficiency of ATP synthesis [ATP produced per oxygen consumed (ATP/O)]. All measurements were performed in samples collected at 3 days of reperfusion from wild-type and DJ-1 KO mice administered AAV9-GFP (GFP) or AAV9-DJ1Δc (DJ1Δc). Values are means ± SEM. 4–7 samples per group. Two-way ANOVA with a Tukey test as the posthoc analysis. **p < 0.01 and ***p < 0.001 vs. wild-type Sham.