Doxorubicin Accumulation in Individually Electrophoresed Organelles

Anderson, Adrian B.; Xiong, Guohua; Arriaga, Edgar A.

doi:10.1021/ja0492539

Cited by 26 publications

(31 citation statements)

References 9 publications

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“…In addition, ABCB8 down-regulation resulted in increased doxorubicin-mediated damage to the mitochondrial DNA [24]. Importantly, doxorubicin was also found to accumulate inside the mitochondria of cell lines [2]. Therefore, it is possible that ABCB8 participates in the export of doxorubicin from the mitochondria, a function that would not be shared with ABCB10.…”

Section: Understanding Abcb10 Function By Phylogenetic Comparison:mentioning

confidence: 99%

Mitochondrial ABC transporters function: The role of ABCB10 (ABC-me) as a novel player in cellular handling of reactive oxygen species

Liesa

Qiu

Shirihai

2012

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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Mitochondria are one of the major sources of reactive oxygen species (ROS) in the cell. When exceeding the capacity of antioxidant mechanisms, ROS production may lead to different pathologies, such as ischemia-reperfusion injury, neurodegeneration, anemia and ageing. As a consequence of the endosymbiotic origin of mitochondria, eukaryotic cells have developed different transport mechanisms that coordinate mitochondrial function with other cellular compartments. Four mitochondrial ATP-binding cassette (ABC) transporters have been described to date in mammals: ABCB6, ABCB8, ABCB7 and ABCB10. ABCB10 is located in the inner mitochondrial membrane forming homodimers, with the ATP binding domain facing the mitochondrial matrix. ABCB10 expression is highly induced during erythroid differentiation and its overexpression increases hemoglobin synthesis in erythroid cells. However, ABCB10 is also expressed in nonerythroid tissues, suggesting a role not directly related to hemoglobin synthesis. Recent evidence points toward ABCB10 as an important player in the protection from oxidative stress in mammals. In this regard, ABCB10 is required for normal erythropoiesis and cardiac recovery after ischemia-reperfusion, processes intimately related to mitochondrial ROS generation. Here, we review the current knowledge on mitochondrial ABC transporters and ABCB10 and discuss the potential mechanisms by which ABCB10 and its transport activity may regulate oxidative stress. We discuss ABCB10 as a potential therapeutic target for diseases in which increased mitochondrial ROS production and oxidative stress play a major role.

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Section: Understanding Abcb10 Function By Phylogenetic Comparison:mentioning

confidence: 99%

Mitochondrial ABC transporters function: The role of ABCB10 (ABC-me) as a novel player in cellular handling of reactive oxygen species

Liesa

Qiu

Shirihai

2012

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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“…Peak analysis has been previously described [40]. Briefly, peak intensities and migration times for individual events with intensities higher than a selected threshold were determined using Pickpeaks, an in-house written Igor Procedure [41]. The rate of appearance of events in the premigration window (false-positive events/s) was used as a correction factor in the migration windows and consequently for estimating the number of true positives.…”

Section: Discussionmentioning

confidence: 99%

CE analysis of the acidic organelles of a single cell

2007

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CE analysis of the acidic organelles of a single cellThe properties of organelles within a cell have been shown to be highly heterogeneous. Until now, it has been unclear just how much of this heterogeneity is endemic to the organelle subpopulations themselves and how much is actually due to stochastic cellular noise. An attractive approach for investigating the origins of heterogeneity among the organelles of a single cell is CE with LIF detection (CE-LIF). As a proof of principle, in this report we optimize and use a single cell CE-LIF method to investigate the properties of endocytic (acidic) organelles. Our results show that the properties of individual acidic organelles containing Alexa Fluor ® 488 Dextran suggest that there are two groups of CCRF-CEM cells: a group with a high dextran content per cell, and a group with a low dextran content per cell. Furthermore, the individual organelle measurements of the single cells allow us to compare in each group the distributions of doxorubicin content per acidic organelle and electrophoretic mobilities of these organelles. Keywords IntroductionHeterogeneity among cells of the same type is known to result partly from intracellular processes, such as pinocytosis [1], the expression of proteins participating in DNA replication [2], and cellular responses to drug treatments [3], and partly from the microenvironment in which the cells reside [4]. Within the cell, heterogeneity at the subcellular level, which refers to variations in the properties of organelles of a given type, has remained practically uncharacterized up to this point. There is a great need to develop and apply single cell approaches capable of measuring the different facets of cellular heterogeneity at the subcellular level. Among the existing techniques for investigating subcellular heterogeneity, chemical imaging has been used to probe the heterogeneity of subcellular properties such as organellar pH, ion concentration, and protein interactions [5,6]. Flow cytometry [7] and CE with LIF detection (CE-LIF) [7,8] have both been used to analyze individual organelles in samples prepared from thousands to millions of cells. With all of these techniques, it has been impossible to track down which organelles originate from the same cell, information that is needed if we are to determine cell-to-cell variations.In order to determine cell-to-cell variations, single cells have been analyzed by CE [9], chromatography [10], MS [11], enzymatic radiolabeling [12], sensitive voltammetric detection [13], and flow cytometry [14]. CE has been used to describe the contents and functions of single cells (e.g., protein expression [15], release of neurotransmitters, and cell secretion [16]). In regard to the subcellular analysis of single cells, CE has been used to sample small subcellular regions, and to measure properties of the whole nucleus of a single cell [17,18]. The ability to detect individual mitochondrial particles by CE-LIF after their release from single cells has been recently described [19]. This report sho...

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“…A 505 nm long-pass filter (505 AELP, Omega Optical, Brattleboro, VT, USA) and a 1.4 mm pinhole were used to reduce light scattering. As previously reported, a dual channel fluorescence detector was used 14. The fluorescence spectral range was split by a 580 nm short-pass dichroic mirror (CP-RR-580, CVI Laser, Albuquerque, NM, USA) and then the fluorescence from Dox was selected by a 635 ± 27.5 nm band-pass filter (XF3015, Omega Optical, Brattleboro, VT, USA) while the fluorescein fluorescence was selected by a 535 ± 17.5 nm band-pass filter (XF3007, Omega Optical, Brattleboro, VT, USA).…”

Section: Methodsmentioning

confidence: 99%

Direct Sampling from Human Liver Tissue Cross Sections for Electrophoretic Analysis of Doxorubicin

et al. 2009

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After chemoembolization of the liver with doxorubicin (Dox), this drug and its metabolites are not homogeneously distributed in this organ. The distribution cannot be easily measured making it difficult to assess how the drug performs in different tissue regions. Here we report a technique for sampling tissue cross-sections that can analyze the contents of micrometer size regions. The tissue cross-sections were from the explanted liver of a hepatocellular carcinoma patient. Samples were directly aspirated from a 5-μm thick tissue cross-section into a 50-μm i.d. capillary where the tissue was solubilized with a separation buffer containing sodium dodecyl sulfate. Upon sample dissolution, Dox and natively fluorescent compounds were separated and detected by micellar electrokinetic chromatography with laser-induced fluorescence detection. Sampling reproducibility and recovery were assessed using 10% (w/v) gelatin as tissue-mimic. Sampling from gelatin slices containing Dox revealed a relative standard deviation of 13%, which was comparable to that of sampling from solution. Dox recovery was 82 ± 16% (n=5). When sampling tumor and non-tumor tissue regions, samples could be taken in the same region 100 μm apart. Atomic force microscopy was used to determine that each sample was 8.4 ± 1.0 pL in volume which made it possible to determine Dox concentrations in the ranges of 0.4 - 1.3 μM and 0.3 - 0.5 μM for the samples taken from tumor and non-tumor regions. The results demonstrated the feasibility of sampling, detection and quantification of Dox in micrometer size regions, which could be a useful resource for analyzing the Dox concentration and distribution in highly heterogeneous tissues.

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Doxorubicin Accumulation in Individually Electrophoresed Organelles

Cited by 26 publications

References 9 publications

Mitochondrial ABC transporters function: The role of ABCB10 (ABC-me) as a novel player in cellular handling of reactive oxygen species

Mitochondrial ABC transporters function: The role of ABCB10 (ABC-me) as a novel player in cellular handling of reactive oxygen species

CE analysis of the acidic organelles of a single cell

Direct Sampling from Human Liver Tissue Cross Sections for Electrophoretic Analysis of Doxorubicin

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