Use of the fluorescent dye 10-N-nonyl acridine orange in quantitative and location assays of cardiolipin: a study on different experimental models

Fernandez, Maria Inmaculada Garcia; Ceccarelli, Daniela; Muscatello, Umberto

doi:10.1016/j.ab.2004.01.020

Cited by 72 publications

(57 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We and others (Mattiazzi et al, 2002) showed increased lipid peroxidation in mitochondrial of G93A-SOD1 mice. We also showed that cardiolipin had a reduced capacity to bind NAO, reflecting peroxidation or loss of cardiolipin (Garcia Fernandez et al, 2004), either of which would compromise the binding of cytochrome c to cardiolipin (Paradies et al, 2000) and would be a prelude to apoptosis (Kirkland et al, 2002;Iverson and Orrenius, 2004;Nakagawa, 2004). Several studies demonstrated that cytochrome c release from mitochondria during apoptosis is a two-step process consisting of the dissociation of this protein from cardiolipin, followed by permeabilization of the outer membrane (McMillin and Dowhan, 2002;Petrosillo et al, 2003;Yuan et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

CytochromecAssociation with the Inner Mitochondrial Membrane Is Impaired in the CNS of G93A-SOD1 Mice

Kirkinezos¹,

et al. 2005

View full text Add to dashboard Cite

A "gain-of-function" toxic property of mutant Cu-Zn superoxide dismutase 1 (SOD1) is involved in the pathogenesis of some familial cases of amyotrophic lateral sclerosis (ALS). Expression of a mutant form of the human SOD1 gene in mice causes a degeneration of motor neurons, leading to progressive muscle weakness and hindlimb paralysis. Transgenic mice overexpressing a mutant human SOD1 gene (G93A-SOD1) were used to examine the mitochondrial involvement in familial ALS. We observed a decrease in mitochondrial respiration in brain and spinal cord of the G93A-SOD1 mice. This decrease was significant only at the last step of the respiratory chain (complex IV), and it was not observed in transgenic wild-type SOD1 and nontransgenic mice. Interestingly, this decrease was evident even at a very early age in mice, long before any clinical symptoms arose. The effect seemed to be CNS specific, because no decrease was observed in liver mitochondria. Differences in complex IV respiration between brain mitochondria of G93A-SOD1 and control mice were abolished when reduced cytochrome c was used as an electron donor, pinpointing the defect to cytochrome c. Submitochondrial studies showed that cytochrome c in the brain of G93A-SOD1 mice had a reduced association with the inner mitochondrial membrane (IMM). Brain mitochondrial lipids, including cardiolipin, had increased peroxidation in G93A-SOD1 mice. These results suggest a mechanism by which mutant SOD1 can disrupt the association of cytochrome c with the IMM, thereby priming an apoptotic program.

show abstract

Section: Discussionmentioning

confidence: 99%

CytochromecAssociation with the Inner Mitochondrial Membrane Is Impaired in the CNS of G93A-SOD1 Mice

Kirkinezos¹,

et al. 2005

View full text Add to dashboard Cite

show abstract

“…If damaged mitochondria are not disposed of in time, it results in oxidation of the cardiolipins in the inner mitochondrial membrane, detected by N-nonyl-acridine orange (NaO) (24,25). Mitochondrial superoxides (detected by MitoSOX) generated during normal oxygen phosphorylation constitute the major ROS in mitochondria and are responsible for cardiolipin oxidation (25).…”

Section: Resultsmentioning

confidence: 99%

Loss of autophagy in erythroid cells leads to defective removal of mitochondria and severe anemia in vivo

Mortensen

Ferguson

Edelmann

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

328

323

View full text Add to dashboard Cite

Timely elimination of damaged mitochondria is essential to protect cells from the potential harm of disordered mitochondrial metabolism and release of proapoptotic proteins. In mammalian red blood cells, the expulsion of the nucleus followed by the removal of other organelles, such as mitochondria, are necessary differentiation steps. Mitochondrial sequestration by autophagosomes, followed by delivery to the lysosomal compartment for degradation (mitophagy), is a major mechanism of mitochondrial turnover. Here we show that mice lacking the essential autophagy gene Atg7 in the hematopoietic system develop severe anemia. Atg7 −/− erythrocytes accumulate damaged mitochondria with altered membrane potential leading to cell death. We find that mitochondrial loss is initiated in the bone marrow at the Ter119 + /CD71 High stage. Proteomic analysis of erythrocyte ghosts suggests that in the absence of autophagy other cellular degradation mechanisms are induced. Importantly, neither the removal of endoplasmic reticulum nor ribosomes is affected by the lack of Atg7. Atg7 deficiency also led to severe lymphopenia as a result of mitochondrial damage followed by apoptosis in mature T lymphocytes. Ex vivo short-lived hematopoietic cells such as monocytes and dendritic cells were not affected by the loss of Atg7. In summary, we show that the selective removal of mitochondria by autophagy, but not other organelles, during erythropoeisis is essential and that this is a necessary developmental step in erythroid cells.Atg7 | mitophagy | lymphopenia | cell death | reactive oxygen species

show abstract

“…The fluorescent probe Mitotracker Green FM (Molecular Probes) binds to mitochondrial membrane lipids regardless of mitochondrial membrane potential or oxidant status (25). Although nonyl acridine orange has been used to determine mitochondrial mass (26), binding may be influenced by altered mitochondrial respiratory states (27). To determine levels of mitochondria within each clone (28), a suspension of 1 Â 10 6 cells/mL in PBS was loaded with 250 nmol/L Mitotracker Green FM in DMSO for 30 minutes at 34jC.…”

Section: Methodsmentioning

confidence: 99%

A Role for Mitochondrial Dysfunction in Perpetuating Radiation-Induced Genomic Instability

Fiskum¹,

Morgan

2006

Cancer Research

140

View full text Add to dashboard Cite

Radiation-induced genomic instability (RIGI) manifests as a heritable increased rate of genetic alterations in the progeny of irradiated cells generations after the initial insult. The progeny can show an increased frequency of chromosomal translocations, deletions, mutations, micronuclei, and decreased plating efficiency. What perpetuates RIGI is unclear; however, persistently increased levels of reactive oxygen species (ROS) are frequently associated with genomically unstable clones. Furthermore, addition of free radical scavengers (e.g., DMSO, glycerol, and cationic thiol cysteamine) reduces the incidence of instability after irradiation, implicating a ROS-mediated role in RIGI induction. Because mitochondria are a major natural cellular source of ROS, we tested the hypothesis that mitochondrial dysfunction has a role in maintaining the elevated ROS levels in our irradiated, genetically unstable GM10115 Chinese hamster ovary cells. Amplex Red fluorometry measurements indicate that the relative contribution of uncoupler-sensitive mitochondrial hydrogen peroxide production to total cellular hydrogen peroxide generation is greater in unstable cells. Measurements of mitochondrial DNA levels and cell cytometric fluorescent measurements of Mitotracker Green FM indicate that differences in mitochondrial ROS production are not due to varying mitochondrial levels. However, mitochondrial respiration measured in digitonin-permeabilized cells is impaired in unstable clones. In addition, manganese superoxide dismutase, a major mitochondrial antioxidant enzyme, exhibits increased immunoreactivity but decreased enzyme activity in unstable clones, which along with decreased respiration rates may explain the increased levels of cellular ROS. These studies show that mitochondria from unstable cells are abnormal and likely contribute to the persistent oxidative stress in the unstable clones. (Cancer Res 2006; 66(21): 10377-83)

show abstract

Use of the fluorescent dye 10-N-nonyl acridine orange in quantitative and location assays of cardiolipin: a study on different experimental models

Cited by 72 publications

References 23 publications

CytochromecAssociation with the Inner Mitochondrial Membrane Is Impaired in the CNS of G93A-SOD1 Mice

CytochromecAssociation with the Inner Mitochondrial Membrane Is Impaired in the CNS of G93A-SOD1 Mice

Loss of autophagy in erythroid cells leads to defective removal of mitochondria and severe anemia in vivo

A Role for Mitochondrial Dysfunction in Perpetuating Radiation-Induced Genomic Instability

Contact Info

Product

Resources

About