Journal: The Journal of Neuroscience

Article Title: Complete Rescue of Cerebrovascular Function in Aged Alzheimer's Disease Transgenic Mice by Antioxidants and Pioglitazone, a Peroxisome Proliferator-Activated Receptor γ Agonist

doi: 10.1523/JNEUROSCI.3348-08.2008

Figure Lengend Snippet: Improved cerebrovascular reactivity coincides with alterations in oxidative stress regulation. A, The responses of isolated arteries to ACh, CGRP, and NOS inhibition with l-NNA (10−5 m) of aged APP mice (▲) relative to wild-type (●) littermates (★p < 0.05, ★★p < 0.01, ★★★p < 0.001) were completely normalized in tempol-, NAC-, and pioglitazone (pio)-treated APP (△) mice, whereas they were unaffected in treated wild-type (○) mice (untreated vs treated APP mice, *p < 0.05, **p < 0.01, ***p < 0.001; n = 4–8 mice/group). B, Pioglitazone, but not NAC or tempol (data not shown), reversed the SOD2 upregulation detected by Western blot in pial vessels of APP mice. Actin was used to normalize loading variation (n = 5–7 mice/group). C, All compounds attenuated the increased SOD2 immunoreactivity detected throughout the cortical neuropil of APP mice, but not the bulk of enzyme immunointensity in neuronal cell bodies (n = 3–5 mice/group). Scale bar, 20 μm. Error bars represent SEM.

Article Snippet: Membranes loaded with proteins (4–20 μg) were incubated with either rabbit anti-SOD1 [1:3000 (vessels)/1:10,000 (cortex); Stressgen], anti-SOD2 (1:20,000/1:10,000; Stressgen), anti-APP (1:2000/1:500; ProSci), anti-BACE1 (1:1000, Santa Cruz Biotechnology), mouse anti-endothelial NOS (eNOS; 1:500; BD Biosciences Transduction Laboratories), anti-cyclooxygenase-2 (COX-2; 1:200/1:100; Cayman), anti-p67 phox (1:200; BD Biosciences Transduction Laboratories), anti-Aβ (6E10, 1:1000/1:200; BioSource International), or anti-actin (1:10,000, Sigma), which was used to normalize loading variation.

Techniques: Isolation, Inhibition, Western Blot

Journal: The Journal of Neuroscience

Article Title: Complete Rescue of Cerebrovascular Function in Aged Alzheimer's Disease Transgenic Mice by Antioxidants and Pioglitazone, a Peroxisome Proliferator-Activated Receptor γ Agonist

doi: 10.1523/JNEUROSCI.3348-08.2008

Figure Lengend Snippet: Treatment effect on amyloidosis. A, APP mice had increased levels of APP in pial vessels (★★★p < 0.001) and cortex (data not shown), although levels of BACE1 were unchanged. They also displayed increased total soluble Aβ, as well as soluble and insoluble Aβ1–40 and Aβ1–42. Neither NAC (data not shown) nor pioglitazone (pio) had an effect on APP, its cleavage enzyme BACE1, or cleavage product Aβ. Actin was used to normalize loading variation (n = 5–7 mice/group). Sol, Soluble; white bars, wild type (WT); light gray bars, WT plus pio; black bars, APP; dark gray bars, APP plus pio. B, Aged APP mice featured extensive Aβ plaque deposition in the cortex and hippocampus (Hi), as evidenced by thioflavine S (left) and Aβ1–42 immunostaining (right), compared with wild-type littermates, which had no deposition (data not shown). The treatments did not affect Aβ plaque number or load (n = 5–6 mice/group). Scale bar, 300 μm. Error bars represent SEM.

Article Snippet: Membranes loaded with proteins (4–20 μg) were incubated with either rabbit anti-SOD1 [1:3000 (vessels)/1:10,000 (cortex); Stressgen], anti-SOD2 (1:20,000/1:10,000; Stressgen), anti-APP (1:2000/1:500; ProSci), anti-BACE1 (1:1000, Santa Cruz Biotechnology), mouse anti-endothelial NOS (eNOS; 1:500; BD Biosciences Transduction Laboratories), anti-cyclooxygenase-2 (COX-2; 1:200/1:100; Cayman), anti-p67 phox (1:200; BD Biosciences Transduction Laboratories), anti-Aβ (6E10, 1:1000/1:200; BioSource International), or anti-actin (1:10,000, Sigma), which was used to normalize loading variation.

Techniques: Immunostaining