Surface Fluorescence Studies of Tissue Mitochondrial Redox State in Isolated Perfused Rat Lungs

Staniszewski, Kevin; Audi, Said H.; Sepehr, Reyhaneh; Jacobs, Elizabeth R.; Ranji, Mahsa

doi:10.1007/s10439-012-0716-z

Cited by 27 publications

(33 citation statements)

References 41 publications

(66 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…; Staniszewski et al . ). During the course of the experiment, the redox ratios do tend generally to decline (repeated measures ANOVA, P < 0.0001), although there were no differences between young and aged (Fig.…”

Section: Resultsmentioning

confidence: 97%

See 1 more Smart Citation

Ageing‐related changes in GABAergic inhibition in mouse auditory cortex, measured using in vitro flavoprotein autofluorescence imaging

Stebbings

Choi

Ravindra

et al. 2015

The Journal of Physiology

View full text Add to dashboard Cite

Key pointsr Ageing is associated with hearing loss and changes in GABAergic signalling in the auditory system.r We tested whether GABAergic signalling in an isolated forebrain preparation also showed ageing-related changes.r A novel approach was used, whereby population imaging was coupled to quantitative pharmacological sensitivity.r Sensitivity to GABA A blockade was inversely associated with age and cortical thickness, but hearing loss did not independently contribute to the change in GABA A ergic sensitivity.r Redox states in the auditory cortex of young and aged animals were similar, suggesting that the differences in GABA A ergic sensitivity are unlikely to be due to differences in slice health.Abstract To examine ageing-related changes in the earliest stages of auditory cortical processing, population auditory cortical responses to thalamic afferent stimulation were studied in brain slices obtained from young and aged CBA/CAj mice (up to 28 months of age). Cortical responses were measured using flavoprotein autofluorescence imaging, and ageing-related changes in inhibition were assessed by measuring the sensitivity of these responses to blockade of GABA A receptors using bath-applied SR95531. The maximum auditory cortical response to afferent stimulation was not different between young and aged animals under control conditions, but responses to afferent stimulation in aged animals showed a significantly lower sensitivity to GABA blockade with SR95531. Cortical thickness, but not hearing loss, improved the prediction of all imaging variables when combined with age, particularly sensitivity to GABA blockade for the maximum response. To determine if the observed differences between slices from young and aged animals were due to differences in slice health, the redox state in the auditory cortex was assessed by measuring the FAD+/NADH ratio using fluorescence imaging. We found that this ratio is highly sensitive to known redox stressors such as H 2 O 2 and NaCN; however, no difference was found between young and aged animals. By using a new approach to quantitatively assess pharmacological sensitivity of population-level cortical responses to afferent stimulation, these data demonstrate that auditory cortical inhibition diminishes with ageing. Furthermore, these data establish a significant relationship between cortical thickness and GABAergic sensitivity, which had not previously been observed in an animal model of ageing.

show abstract

Section: Resultsmentioning

confidence: 97%

“…; Staniszewski et al . ). We also determined that redox ratio is a sensitive marker of tissue oxidative stress, as low concentrations of H 2 O 2 and NaCN induce dose‐dependent changes in redox signal, as seen previously (Gerich et al .…”

Section: Discussionmentioning

confidence: 97%

Ageing‐related changes in GABAergic inhibition in mouse auditory cortex, measured using in vitro flavoprotein autofluorescence imaging

Stebbings

Choi

Ravindra

et al. 2015

The Journal of Physiology

View full text Add to dashboard Cite

show abstract

“…We have recently reported this technique for the determination of the oxidative state of the kidney in SS rats 28 . The frozen kidneys, stored at −80°C until the day of study, were mounted in the cryoimager, a custom designed and fabricated optical fluorescence imaging system from the Biophotonics Lab at the University of Wisconsin-Milwaukee previously described in detail 28,38,39 .…”

Section: Methodsmentioning

confidence: 99%

Evidence of the Importance of Nox4 in Production of Hypertension in Dahl Salt-Sensitive Rats

et al. 2016

Self Cite

View full text Add to dashboard Cite

This study reports the consequences of knocking out NADPH oxidase 4 (Nox4) upon the development of hypertension and kidney injury in the Dahl salt-sensitive (SS) rat. Zinc finger nuclease injection of single cell SS embryos was used to create an 8 base-pair frame-shift deletion of Nox4 resulting in a loss of the ~68 kD band in Western blot analysis of renal cortical tissue of the SSNox4−/− rats. SSNox4−/− rats exhibited a significant reduction of salt-induced hypertension compared to SS rats after 21 days of 4.0% NaCl diet (134±5 vs 151±3 mmHg in SS) and a significant reduction of albuminuria, tubular casts, and glomerular injury. Optical fluorescence 3D cryoimaging revealed significantly higher redox ratios (NADH/FAD) in the kidneys of SSNox4−/− rats even when fed the 0.4% NaCl diet indicating greater levels of mitochondrial electron transport chain metabolic activity and reduced oxidative stress compared to SS rats. Prior to the development of hypertension, RNA expression levels of NADPH oxidase subunits Nox2, p67phox, and p22phox were found to be significantly lower (p<0.05) in SSNox4−/− compared to SS rats in the renal cortex. Thus the mutation of Nox4 appears to modify transcription of a number of genes in ways that contribute to the protective effects observed in the SSNox4−/− rats. We conclude that the reduced renal injury and attenuated blood pressure response to high salt in the SSNox4−/− rat could be the result of multiple pathways including gene transcription, mitochondrial energetics, oxidative stress, and protein matrix production impacted by the knock out of Nox4.

show abstract

“…Evaluation of redox state of pyridine nucleotides is widely used in biochemical research, in particular in studies on consequences of oxidative stress [11][12][13], cell death detection [8], estimation of level of postsynaptic neuronal activation [10,14]. The corresponding experiments were made on carcinoma cells [15], lung cells [16], heart tissue [17][18][19]. Changes in redox state of pyridine nucleotides as the primary cause of disruptions in mitochondrial respiration under fluoroacetate toxicity have been studied on ascitic Ehrlich carcinoma and rat liver mitochondria [20].…”

mentioning

confidence: 99%

Evaluation of functioning of mitochondrial electron transport chain with NADH and FAD autofluorescence

Danylovych¹

2016

Ukr.Biochem.J.

View full text Add to dashboard Cite

We prove the feasibility of evaluation of mitochondrial electron transport chain function in isolated mitochondria of smooth muscle cells of rats from uterus using fluorescence of NADH and FAD coenzymes. We found the inversely directed changes in FAD and NADH fluorescence intensity under normal functioning of mitochondrial electron transport chain. The targeted effect of inhibitors of complex I, III and IV changed fluorescence of adenine nucleotides. Rotenone (5 μM) induced rapid increase in NADH fluorescence due to inhibition of complex I, without changing in dynamics of FAD fluorescence increase. Antimycin A, a complex III inhibitor, in concentration of 1 μg/ml caused sharp increase in NADH fluorescence and moderate increase in FAD fluorescence in comparison to control. NaN3 (5 mM), a complex IV inhibitor, and CCCP (10 μM), a protonophore, caused decrease in NADH and FAD fluorescence. Moreover, all the inhibitors caused mitochondria swelling. NO donors, e.g. 0.1 mM sodium nitroprusside and sodium nitrite similarly to the effects of sodium azide. Energy-dependent Ca2+ accumulation in mitochondrial matrix (in presence of oxidation substrates and Mg-ATP2- complex) is associated with pronounced drop in NADH and FAD fluorescence followed by increased fluorescence of adenine nucleotides, which may be primarily due to Ca2+- dependent activation of dehydrogenases of citric acid cycle. Therefore, the fluorescent signal of FAD and NADH indicates changes in oxidation state of these nucleotides in isolated mitochondria, which may be used to assay the potential of effectors of electron transport chain.

show abstract

Surface Fluorescence Studies of Tissue Mitochondrial Redox State in Isolated Perfused Rat Lungs

Cited by 27 publications

References 41 publications

Ageing‐related changes in GABAergic inhibition in mouse auditory cortex, measured using in vitro flavoprotein autofluorescence imaging

Ageing‐related changes in GABAergic inhibition in mouse auditory cortex, measured using in vitro flavoprotein autofluorescence imaging

Evidence of the Importance of Nox4 in Production of Hypertension in Dahl Salt-Sensitive Rats

Evaluation of functioning of mitochondrial electron transport chain with NADH and FAD autofluorescence

Contact Info

Product

Resources

About