Labeling Mitochondria with JC-1

Chazotte, Brad

doi:10.1101/pdb.prot065490

Cited by 83 publications

(69 citation statements)

References 3 publications

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“…4B). These findings were confirmed by measurement of the cationic dye JC-1, which forms red-fluorescent J-aggregates at normal/high DCm and green-fluorescent monomers upon depolarization of the DCm (Chazotte, 2011). AG311 resulted in a decrease in the ratio of aggregates (red) over monomers (green) in MDA-MB-435 cells at 15 minutes post treatment (Fig.…”

Section: Ag311 Induces Rapid Mitochondrial Membranesupporting

confidence: 62%

A Small Molecule with Anticancer and Antimetastatic Activities Induces Rapid Mitochondrial-Associated Necrosis in Breast Cancer

Bastian

Thorpe

Disch

et al. 2015

J Pharmacol Exp Ther

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Therapy for treatment-resistant breast cancer provides limited options and the response rates are low. Therefore, the development of therapies with alternative chemotherapeutic strategies is necessary. AG311 (5-[(4-methylphenyl)thio]-9H-pyrimido [4,5-b]indole-2,4-diamine), a small molecule, is being investigated in preclinical and mechanistic studies for treatment of resistant breast cancer through necrosis, an alternative cell death mechanism. In vitro, AG311 induces rapid necrosis in numerous cancer cell lines as evidenced by loss of membrane integrity, ATP depletion, HMGB1 (high-mobility group protein B1) translocation, nuclear swelling, and stable membrane blebbing in breast cancer cells. Within minutes, exposure to AG311 also results in mitochondrial depolarization, superoxide production, and increased intracellular calcium levels. Additionally, upregulation of mitochondrial oxidative phosphorylation results in sensitization to AG311. This AG311-induced cell death can be partially prevented by treatment with the mitochondrial calcium uniporter inhibitor, Ru360 [(m)[(HCO 2 )(NH 3 ) 4 Ru] 2 OCl 3 ], or an antioxidant, lipoic acid. Additionally, AG311 does not increase apoptotic markers such as cleavage of poly (ADP-ribose) polymerase (PARP) or caspase-3 and -7 activity. Importantly, in vivo studies in two orthotopic breast cancer mouse models (xenograft and allograft) demonstrate that AG311 retards tumor growth and reduces lung metastases better than clinically used agents and has no gross or histopathological toxicity. Together, these data suggest that AG311 is a first-in-class antitumor and antimetastatic agent inducing necrosis in breast cancer tumors, likely through the mitochondria.

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Section: Ag311 Induces Rapid Mitochondrial Membranesupporting

confidence: 62%

A Small Molecule with Anticancer and Antimetastatic Activities Induces Rapid Mitochondrial-Associated Necrosis in Breast Cancer

Bastian

Thorpe

Disch

et al. 2015

J Pharmacol Exp Ther

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“…To detect changes in mitochondrial membrane potential, we used JC-1 probe that shows dual emission (based on the creation of aggregates on monomers in dependence on mitochondrial membrane potential). This probe is usually used in animal cell models (Binet et al, 2014;Chazotte, 2011), however, its application in plant (cell) models has been also described (Simeonova et al, 2004). In our case, we observed only limited creation of JC-1 aggregates in juglone-treated lettuce roots, which indicates decreasing mitochondrial potential.…”

Section: Discussionmentioning

confidence: 70%

Phytotoxic action of naphthoquinone juglone demonstrated on lettuce seedling roots

Babula

Vaverkova

Pobořilová

et al. 2014

Plant Physiology and Biochemistry

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“…Use of a probe that monitors inner mitochondrial ΔΨm can be used as a readout for mitochondrial health. JC-1 is a cationic fluorescent dye whose accumulation is potential-dependent (Chazotte, 2011). The monomeric form of JC-1 emits at ~529 nm (green) and accumulates proportionally with the strength of the membrane potential.…”

Section: Mitochondria In Drosophila Muscle Tissuementioning

confidence: 99%

Analysis of mitochondrial structure and function in the Drosophila larval musculature

2016

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Mitochondria are dynamic organelles that change their architecture in normal physiological conditions. Mutations in genes that control mitochondrial fission or fusion, such as dynamin-related protein (Drp1), Mitofusins 1 (Mfn1) and 2 (Mfn2), and Optic atrophy 1 (Opa1), result in neuropathies or neurodegenerative diseases. It is increasingly clear that altered mitochondrial dynamics also underlie the pathology of other degenerative diseases, including Parkinson’s disease (PD). Thus, understanding mitochondrial distribution, shape, and dynamics in all cell types is a prerequisite for developing and defining treatment regimens that may differentially affect tissues. The majority of Drosophila genes implicated in mitochondrial dynamics have been studied in the adult indirect flight muscle (IFM). Here, we discuss the utility of Drosophila third instar larvae (L3) as an alternative model to analyze and quantify mitochondrial behaviors. Advantages include large muscle cell size, a stereotyped arrangement of mitochondria that is conserved in mammalian muscles, and the ability to analyze muscle-specific gene function in mutants that are lethal prior to adult stages. In particular, we highlight methods for sample preparation and analysis of mitochondrial morphological features.

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Labeling Mitochondria with JC-1

Cited by 83 publications

References 3 publications

A Small Molecule with Anticancer and Antimetastatic Activities Induces Rapid Mitochondrial-Associated Necrosis in Breast Cancer

A Small Molecule with Anticancer and Antimetastatic Activities Induces Rapid Mitochondrial-Associated Necrosis in Breast Cancer

Phytotoxic action of naphthoquinone juglone demonstrated on lettuce seedling roots

Analysis of mitochondrial structure and function in the Drosophila larval musculature

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