Labeling Mitochondria with MitoTracker Dyes

Chazotte, Brad

doi:10.1101/pdb.prot5648

Cited by 157 publications

(109 citation statements)

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“…The Δψm is an important parameter of mitochondrial function and may be used as an indicator of cell growth (16). A high Δψm has been observed in rapidly proliferating cells with complex form called J-aggregate, while a low Δψm has been observed in apoptotic cells with a monomeric form of JC-1 (17). In the current study, a significant decrease in Δψm was detected following treatment with BDMC for 24 h. ROS are chemically reactive molecules, which at low levels, facilitate cancer cell survival by cell-cycle promotion; however, at higher levels may suppress tumor growth via the activation of cell-cycle inhibitors and the induction of cell death (18).…”

Section: Discussionmentioning

confidence: 99%

Bisdemethoxycurcumin attenuates gastric adenocarcinoma growth by inducing mitochondrial dysfunction

Luo

Wei

et al. 2014

Oncology Letters

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Bisdemethoxycurcumin (BDMC) is a demethoxy derivative of curcumin. In this study, a human gastric adenocarcinoma xenograft model was generated in vivo using nude mice and BDMC was observed to suppress the growth and activity of tumors, in addition to improving the physical and mental capacity of the mice. An increased number of apoptotic cells, decreased ratio of B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein and increased caspase-3 expression was also observed following treatment with BDMC, indicating that BDMC may promote apoptosis in tumors via mitochondrial modulation. The growth of SGC 7901 gastric cancer cells was inhibited and arrested at G1 phase. Specific indicators of mitochondrial dysfunction, a reduction in adenosine triphosphate generation, the inner mitochondrial membrane potential, augmentation of reactive oxygen species production and cytochrome c were also detected in the mitochondria following treatment with BDMC. These results indicate that BDMC attenuates gastric adenocarcinoma growth by inducing mitochondrial dysfunction.

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Section: Discussionmentioning

confidence: 99%

Bisdemethoxycurcumin attenuates gastric adenocarcinoma growth by inducing mitochondrial dysfunction

Luo

Wei

et al. 2014

Oncology Letters

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“…Cells were plated at 1 x 10 6 cells/mL in RPMI complete medium for 24 h. Samples were labeled with Rh123 (#R8004, Sigma-Aldrich, 10 μg/mL) for 15 min at 37°C, washed, and MFI was measured by flow cytometry using λ ex = 488 nm and λ em = 530/30 nm bandpass filter (Rh123) [50]. …”

Section: Methodsmentioning

confidence: 99%

Mitochondrial-Targeted Decyl-Triphenylphosphonium Enhances 2-Deoxy-D-Glucose Mediated Oxidative Stress and Clonogenic Killing of Multiple Myeloma Cells

et al. 2016

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Therapeutic advances have markedly prolonged overall survival in multiple myeloma (MM) but the disease currently remains incurable. In a panel of MM cell lines (MM.1S, OPM-2, H929, and U266), using CD138 immunophenotyping, side population staining, and stem cell-related gene expression, we demonstrate the presence of stem-like tumor cells. Hypoxic culture conditions further increased CD138low stem-like cells with upregulated expression of OCT4 and NANOG. Compared to MM cells, these stem-like cells maintained lower steady-state pro-oxidant levels with increased uptake of the fluorescent deoxyglucose analog. In primary human MM samples, increased glycolytic gene expression correlated with poorer overall and event-free survival outcomes. Notably, stem-like cells showed increased mitochondrial mass, rhodamine 123 accumulation, and orthodox mitochondrial configuration while more condensed mitochondria were noted in the CD138high cells. Glycolytic inhibitor 2-deoxyglucose (2-DG) induced ER stress as detected by qPCR (BiP, ATF4) and immunoblotting (BiP, CHOP) and increased dihydroethidium probe oxidation both CD138low and CD138high cells. Treatment with a mitochondrial-targeting agent decyl-triphenylphosphonium (10-TPP) increased intracellular steady-state pro-oxidant levels in stem-like and mature MM cells. Furthermore, 10-TPP mediated increases in mitochondrial oxidant production were suppressed by ectopic expression of manganese superoxide dismutase. Relative to 2-DG or 10-TPP alone, 2-DG plus 10-TPP combination showed increased caspase 3 activation in MM cells with minimal toxicity to the normal hematopoietic progenitor cells. Notably, treatment with polyethylene glycol conjugated catalase significantly reduced 2-DG and/or 10-TPP-induced apoptosis of MM cells. Also, the combination of 2-DG with 10-TPP decreased clonogenic survival of MM cells. Taken together, this study provides a novel strategy of metabolic oxidative stress-induced cytotoxicity of MM cells via 2-DG and 10-TPP combination therapy.

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“…Once one is experienced with the use of dyes another approach is to utilize confocal microscopy with Z-stacks to image only mitochondria in the right tissue; the mitochondrial networks in muscle are quite distinct compared to other tissues. Unfortunately, the inability of C 32 H 32 Cl 2 N 2 O to leave mitochondria 28 means that, unlike the sarcomere and mitochondrial imaging techniques discussed above, it cannot be used to prospectively follow loss of mitochondrial function with time, unless C 32 H 32 Cl 2 N 2 O is added at discrete time points to separate animal populations. This limitation can be overcome by using dyes such as JC-10 that do leave the mitochondria upon collapse of membrane potential 43 .…”

Section: Imaging Of Mitochondrial Networkmentioning

confidence: 99%

“…As mitochondrial structural disruptions may or may not alter mitochondrial functional capacity, it may be desirable to assess mitochondrial function in animals displaying altered mitochondrial structure. C 32 H 32 Cl 2 N 2 O accumulates in mitochondria, of all cell types, where a membrane potential is present and stains the mitochondria 28 ( Figure 4C). To specifically assess mitochondrial function in muscle, one can use C 32 H 32 Cl 2 N 2 O and GFP labeled muscle mitochondria to demonstrate the effects of an intervention specifically upon muscle mitochondria ( Figure 4C).…”

Section: Assessment Of Mitochondrial Function In Vivomentioning

confidence: 99%

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Methods to Assess Subcellular Compartments of Muscle in <em>C. elegans</em>

Gaffney

Bass

Barratt

et al. 2014

JoVE

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Muscle is a dynamic tissue that responds to changes in nutrition, exercise, and disease state. The loss of muscle mass and function with disease and age are significant public health burdens. We currently understand little about the genetic regulation of muscle health with disease or age. The nematode C. elegans is an established model for understanding the genomic regulation of biological processes of interest. This worm's body wall muscles display a large degree of homology with the muscles of higher metazoan species. Since C. elegans is a transparent organism, the localization of GFP to mitochondria and sarcomeres allows visualization of these structures in vivo. Similarly, feeding animals cationic dyes, which accumulate based on the existence of a mitochondrial membrane potential, allows the assessment of mitochondrial function in vivo. These methods, as well as assessment of muscle protein homeostasis, are combined with assessment of whole animal muscle function, in the form of movement assays, to allow correlation of sub-cellular defects with functional measures of muscle performance. Thus, C. elegans provides a powerful platform with which to assess the impact of mutations, gene knockdown, and/or chemical compounds upon muscle structure and function. Lastly, as GFP, cationic dyes, and movement assays are assessed non-invasively, prospective studies of muscle structure and function can be conducted across the whole life course and this at present cannot be easily investigated in vivo in any other organism.

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Labeling Mitochondria with MitoTracker Dyes

Cited by 157 publications

References 1 publication

Bisdemethoxycurcumin attenuates gastric adenocarcinoma growth by inducing mitochondrial dysfunction

Bisdemethoxycurcumin attenuates gastric adenocarcinoma growth by inducing mitochondrial dysfunction

Mitochondrial-Targeted Decyl-Triphenylphosphonium Enhances 2-Deoxy-D-Glucose Mediated Oxidative Stress and Clonogenic Killing of Multiple Myeloma Cells

Methods to Assess Subcellular Compartments of Muscle in <em>C. elegans</em>

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