Phosphorescence Oxygen Analyzer as a Measuring Tool for Cellular Bioenergetics

Al-Jasmi, Fatma; Suwaidi, Ahmed R. Al; Al-Shamsi, Mariam; Marzouqi, Farida; Mansouri, Aysha Al; Shaban, Sami; Penefsky, Harvey S.; Souid, Abdul‐Kader

doi:10.5772/31409

Cited by 3 publications

(8 citation statements)

References 56 publications

(61 reference statements)

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“…Glucose oxidase (GO; catalogue #G7141) was purchased from Sigma-Aldrich (St. Louis, MO); its solution (10 mg/mL) was dissolved in dH 2 O and stored at -20 C. Potassium cyanide (KCN, catalogue # 10201) was purchased from BDH Middle East LLC (Dubai, UAE); its solution was prepared in dH 2 O immediately prior to use. The remaining reagents were purchased from Sigma-Aldrich and prepared and stored as previously described [17,18,30,31,32,33]. Sodium pentobarbital (90 μg/g, given intraperitoneally) was used for anesthesia [34].…”

Section: Chemicals and Reagentsmentioning

confidence: 99%

“…and various concentrations of β-glucose [16,17,18]. Potassium cyanide (KCN, approximately 10 mM, prepared in dH 2 O) inhibited respiration, confirming oxygen was reduced in the mitochondrial respiratory chain [33].…”

Section: Oxygen Measurementmentioning

confidence: 99%

“…Reported biotechnologies for measuring dissolved oxygen include: ( 1 ) Electrochemical sensors (Clark electrodes, polarographic or Galvanic) that generate electrical signals in response to oxygen reduction by the polarized electrode [ 12 ]; ( 2 ) Optical sensors based on oxygen quenching fluorescence [ 13 , 14 , 15 ], phosphorescence [ 16 , 17 , 18 ], or luminescence; and ( 3 ) Micro-electrochemical techniques (scanning electrochemical microscopy) based on measurements of flow of small molecules, such as oxygen in tissues [ 19 ]. The use of ‘fluorophore-based biosensors, including solid state cartridges’ [ 20 , 21 , 22 ] and ‘high-resolution FluoRespirometry’ [ 14 ] have become relatively popular.…”

Section: Introductionmentioning

confidence: 99%

“…A number of recent neuroscience studies have incorporated cellular respiration as a biomarker for monitoring adverse events [ 23 , 24 , 25 , 26 , 27 , 28 ] or therapeutic interventions, such as effects of cytoprotective therapies [ 29 ]. We have developed and used the previously reported methodology for oxygen measurements via phosphorescence [ 16 ] to study cellular bioenergetics in cell cultures and various tissues [ 17 , 18 ]. The main principle of this analytical tool is that oxygen quenches the phosphorescence of pallidum phosphors, such as Pd(II) meso-tetra (sulfophenyl) tetrabenzoporphyrin [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

“…This analytical tool (phosphorescence oxygen analyzer) has been especially useful in monitoring oxygen reactions of rapid kinetics [ 30 ] as well as oxygen measurements over several hours [ 31 ]. The applied instrument and its software development have been previously described in details [ 18 , 32 ].…”

Section: Introductionmentioning

confidence: 99%

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Conserved spinal cord bioenergetics in experimental autoimmune encephalomyelitis in C57BL6 mice, measured using phosphorescence oxygen analyzer

Shamsi

Shahin

Kamyan

et al. 2021

Heliyon

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Background: We have previously reported that spinal cord respiration (cellular mitochondrial oxygen consumption) and ATP content are conserved in the studied model of experimental autoimmune encephalomyelitis (EAE), foreseeing a recovery of the diseased rats. This exemplary lesion of multiple sclerosis is used here to measure spinal cord bioenergetics in C57BL6 mice. Our hypothesis is that, despite the well-known focal axonal mitochondrial pathology, bioenergetics of the CNS is reasonably preserved in this disease. Methods: EAE was induced with an immunodominant myelin oligodendrocyte glycoprotein epitope in complete Freund's adjuvant, appended by injections of pertussis toxin. A low-and high-dose of the encephalitogen, administered into base of tail or hind-flank, were investigated. Control mice received only the incomplete adjuvant into tail. Oxygen measurements were based on quenching the phosphorescence of Pd(II) meso-tetra (sulfophenyl) tetrabenzoporphyrin by molecular oxygen. Cellular ATP was measured using the luciferin/luciferase system. Results: The kinetics of spinal cord oxygen consumption was zero-order (linear with time) and inhibited by cyanide, confirming oxygen was reduced by cytochrome oxidase. The rate of respiration (in μM O 2 .min À1 .mg À1 ; measured on Days 13-28) in control mice was (mean AE SD) 0.086 AE 0.024 (n ¼ 8) and in immunized mice was 0.079 AE 0.020 (n ¼ 15, P ¼ 0.265, Mann-Whitney test). Consistently, cellular ATP (in μmol mg À1 dry pellet weight; measured on Days 13-28) in control mice was 0.068 AE 0.079 (n ¼ 11) and in immunized mice was 0.063 AE 0.061 (n ¼ 24, P ¼ 0.887, Mann-Whitney U test). Conclusions: In vitro measurements of spinal cord bioenergetics show conservation of the mitochondrial function in mice with EAE. These results suggest the previously documented reduced mitochondrial electrochemical potential in this disease is alterable, and likely reflects the adverse events of neuroinflammation.

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Section: Chemicals and Reagentsmentioning

confidence: 99%

Section: Oxygen Measurementmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Conserved spinal cord bioenergetics in experimental autoimmune encephalomyelitis in C57BL6 mice, measured using phosphorescence oxygen analyzer

Shamsi

Shahin

Kamyan

et al. 2021

Heliyon

Self Cite

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The phosphorescence oxygen analyzer as a screening tool for disorders with impaired lymphocyte bioenergetics

Al-Jasmi

Penefsky

Souid

2011

Molecular Genetics and Metabolism

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In Vitro Study on the Response of Fibroblast Cellular Respiration to Lipoic acid, Thiamine and Carnitine in Patients with Dihydrolipoyl Dehydrogenase Deficiency

Al-Jasmi

Pramathan²,

Kowash³

et al. 2016

J Mol Genet Med

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Objectives: This study examined in vitro responses of fibroblast cellular respiration to lipoic acid, thiamine and carnitine in patients with dihydrolipoyl dehydrogenase (DLD) deficiency. This disorder impairs cellular bioenergetics and these compounds are used to improve clinical manifestations of the disease. The study aimed to utilize mitochondrial O 2 consumption as a surrogate biomarker for examining cellular responses to metabolic therapies.Methods: Cultured fibroblasts from three patients were treated with therapeutic concentrations of the compounds for 24 hours. Cells were then harvested and processed for measuring respiration using phosphorescence oxygen analyzer. Patients 1 and 2 were severely symptomatic infants with homozygous c.1436A>T mutation in the DLD gene. Patient-3 was a mildly symptomatic adolescent with homozygous c.685G>T mutation. Results:The rate of respiration (mean ± SD, n=6, µM O 2 min -1 /10 7 cells) in fibroblasts from a normal infant was 9.3 ± 1.6, in fibroblasts from Patient-1 was 5.1 ± 0.9 (p=0.001), in fibroblasts from Patient-2 was 7.4 ± 1.4 (p=0.051), and in fibroblasts from Patient-3 was 10.3 ± 3.3 (p=0.836). In normal fibroblasts, respiration decreased by the thiamine (p=0.012) and carnitine (p=0.023) treatments. In Patient-1, respiration increased by the lipoic acid (p<0.002), thiamine (p<0.001), and carnitine (p=0.018) treatments; this patient clinically responded to thiamine. In Patient-2, respiration decreased by the thiamine (p=0.026) and carnitine (p=0.008) treatments; this patient did not respond to these drugs. In Patient-3, respiration increased by the carnitine (p=0.012) treatment; the patient clinically responded to carnitine. Conclusions:The results show cellular respiration is a suitable biomarker for the disease. The significance of using this tool to assess responses to therapies requires further studies.

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Phosphorescence Oxygen Analyzer as a Measuring Tool for Cellular Bioenergetics

Cited by 3 publications

References 56 publications

Conserved spinal cord bioenergetics in experimental autoimmune encephalomyelitis in C57BL6 mice, measured using phosphorescence oxygen analyzer

Conserved spinal cord bioenergetics in experimental autoimmune encephalomyelitis in C57BL6 mice, measured using phosphorescence oxygen analyzer

The phosphorescence oxygen analyzer as a screening tool for disorders with impaired lymphocyte bioenergetics

In Vitro Study on the Response of Fibroblast Cellular Respiration to Lipoic acid, Thiamine and Carnitine in Patients with Dihydrolipoyl Dehydrogenase Deficiency

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