electrocardiogram (ecg), respiration and body temperature monitoring system  (SA Instruments)

Journal: Redox Biology

Article Title: Mitochondrial and redox modifications in early stages of Huntington's disease

doi: 10.1016/j.redox.2022.102424

Figure Lengend Snippet: Oxygen consumption rate (OCR) in skin fibroblasts from premanifest and manifest Huntington's disease carriers and controls. (A – C) OCR of cells treated with the different compounds. The mitochondrial inhibitors were sequentially injected into different ports of the Seahorse XF24 analyzer and the final concentrations of each were: 1 μM oligomycin (O), 0.3 μM FCCP (F), 1 μM rotenone and 1 μM antimycin A (R + A). ( D) (i) Levels of basal OCR; (ii) Oxygen consumed for ATP generation through the complex V; (iii) Maximal respiration capacity; (iv) Spare respiratory capacity; (v) Component of OCR representing passive H + leakage across the mitochondrial inner membrane were calculated as described in Methods section; individual data are presented as floating bars (min, max) with mean lines shown at least three independent experiments; group analysis represents the mean ± SEM values (controls (C, CTR), premanifest (Pre-M) and manifest HD patients) and are plotted on the secondary Y axis (blue). (E) Correlation was performed using the Spearman correlation coefficient ρ (sig; n). Statistical analysis: One-Way ANOVA – post hoc Bonferroni's multiple comparisons test. *p < 0.05 (controls vs premanifest); φφ p<0.01 (controls vs HD manifest) and φ p<0.05, φφ p<0.01, φφφ p<0.001 (premanifest vs HD manifest). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Mice were kept anesthetized by isoflurane (1–2%) with 100% O 2 with body temperature and respiration monitoring (SA Instruments SA, Stony Brook, USA).

Techniques: Injection, Membrane

Journal: Redox Biology

Article Title: Mitochondrial and redox modifications in early stages of Huntington's disease

doi: 10.1016/j.redox.2022.102424

Figure Lengend Snippet: Oxygen consumption rates (OCR) in 3 mo YAC128 mouse striatal and cortical mitochondria. Mitochondrial respiration was measured using the Seahorse flux analyzer in 3 mo YAC128 and WT striatal (A) and cortical (B) isolated mitochondria. Levels of respiratory coupling ( A i and B i ; for striatal and cortical mitochondria, respectively) were analysed in MAS containing 10 mM succinate plus 2 μM rotenone under sequentially injection of mitochondrial inhibitors and substrates (final concentration: 4 mM ADP; 2.5 μg/mL oligomycin; 4 μM FCCP and 4 μM antimycin A) as shown in representative traces and basal respiration and maximal respiration, ATP production and H + calculated as described in Methods section. The activity of mitochondrial respiratory chain complexes ( A ii and B ii ; for striatal and cortical mitochondria, respectively) was analysed in uncoupling conditions performed in MAS containing 4 μM FCCP, 10 mM pyruvate and 2 mM malate. Mitochondrial inhibitors and substrates were sequentially injected (final concentration: 2 μM rotenone, 10 mM succinate, 4 μM antimycin A and 10 mM ascorbate/100 μM TMPD) as shown in the representative traces and Cx I-IV activities calculated as described in Methods section. Data are the mean ± SEM of experiments performed in independent mitochondrial preparations obtained from 3 to 7 mice from each genotype, run in duplicates or triplicates. Statistical analysis: *p < 0.05, **p < 0.01 when compared with WT mitochondria, by nonparametric Mann-Whitney U test.

Article Snippet: Mice were kept anesthetized by isoflurane (1–2%) with 100% O 2 with body temperature and respiration monitoring (SA Instruments SA, Stony Brook, USA).

Techniques: Isolation, Injection, Concentration Assay, Activity Assay, MANN-WHITNEY