Comparison of Mitochondrial Fluorescent Dyes in Stromal Cells

Buravkov, S. V.; Pogodina, M. V.; Буравкова, Л. Б.

doi:10.1007/s10517-014-2637-3

Cited by 11 publications

(8 citation statements)

References 11 publications

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“…Interestingly, GC7 never induced a total breakdown of DC m , which was classically observed in cells entering apoptosis (total DC m dissipation is ,3% of the GC7-treated cells only; area M1) ( Figure 2C). We obtained the same effect by measuring directly the fluorescence of the mitochondrial potential probe Mitotracker red FM 18 on intact confluent epithelia (Supplemental Figure 4). Interestingly, this GC7-induced decrease in DC m was not related to an upregulation of the uncoupling proteins UCP2 (Supplemental Figure 3B) or UCP3 (Supplemental Figure 3C).…”

Section: Inhibition Of Eif5a Hypusination Induces Metabolicmentioning

confidence: 84%

Targeting eIF5A Hypusination Prevents Anoxic Cell Death through Mitochondrial Silencing and Improves Kidney Transplant Outcome

Melis

Rubera

Cougnon

et al. 2016

JASN

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The eukaryotic initiation factor 5A (eIF5A), which is highly conserved throughout evolution, has the unique characteristic of post-translational activation through hypusination. This modification is catalyzed by two enzymatic steps involving deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase (DOHH). Notably, eIF5A may be involved in regulating the lifespan of during long-term hypoxia. Therefore, we investigated the possibility of a link between eIF5A hypusination and cellular resistance to hypoxia/anoxia. Pharmacologic targeting of DHPS by1-guanyl-1,7-diaminoheptane (GC7) or RNA interference-mediated inhibition of DHPS or DOHH induced tolerance to anoxia in immortalized mouse renal proximal cells. Furthermore, GC7 treatment of cells reversibly induced a metabolic shift toward glycolysis as well as mitochondrial remodeling and led to downregulated expression and activity of respiratory chain complexes, features characteristic of mitochondrial silencing. GC7 treatment also attenuated anoxia-induced generation of reactive oxygen species in these cells and in normoxic conditions, decreased the mitochondrial oxygen consumption rate of cultured cells and mice. In rats, intraperitoneal injection of GC7 substantially reduced renal levels of hypusinated eIF5A and protected against ischemia-reperfusion-induced renal injury. Finally, in the preclinical pig kidney transplant model, intravenous injection of GC7 before kidney removal significantly improved graft function recovery and late graft function and reduced interstitial fibrosis after transplant. This unconventional signaling pathway offers an innovative therapeutic target for treating hypoxic-ischemic human diseases and organ transplantation.

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Section: Inhibition Of Eif5a Hypusination Induces Metabolicmentioning

confidence: 84%

Targeting eIF5A Hypusination Prevents Anoxic Cell Death through Mitochondrial Silencing and Improves Kidney Transplant Outcome

Melis

Rubera

Cougnon

et al. 2016

JASN

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“…To assess localization, we compared the intracellular fluorescent emission of 2 and 3 to that of MitoTracker Red CMXRos (MTR), which is known to localize in the mitochondria . Incubation of A549 cells with 2 at 100 μM for 4 h showed that the compound localizes similarly to the MTR dye as indicated by the yellow-orange color in the merged image (Figure ).…”

Section: Resultsmentioning

confidence: 99%

Mitochondrial-Localized Versus Cytosolic Intracellular CO-Releasing Organic PhotoCORMs: Evaluation of CO Effects Using Bioenergetics

et al. 2018

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While interactions between carbon monoxide (CO) and mitochondria have been previously studied, the methods used to deliver CO (gas or CO-releasing metal carbonyl compounds) lack subcellular targeting and/or controlled delivery. Thus, the effective concentration needed to produce changes in mitochondrial bioenergetics is yet to be fully defined. To evaluate the influence of mitochondrial-targeted versus intracellularly released CO on mitochondrial oxygen consumption rates, we developed and characterized flavonol-based CO donor compounds that differ at their site of release. These molecules are metal-free, visible light triggered CO donors (photoCORMs) that quantitatively release CO and are trackable in cells via confocal microscopy. Our studies indicate that at a concentration of 10 μM, the mitochondrial-localized and cytosolic CO-releasing compounds are similarly effective in terms of decreasing ATP production, maximal respiration, and the reserve capacity of A549 cells. This concentration is the lowest to impart changes in mitochondrial bioenergetics for any CO-releasing molecule (CORM) reported to date. The results reported herein demonstrate the feasibility of using a structurally tunable organic photoCORM framework for comparative intracellular studies of the biological effects of carbon monoxide.

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“…However, according to the dye's covalent binding to sulphhydryl groups, it may only tentatively be considered as potential sensitive mitochondrial dye, and we cannot discern whether the activity or the number of mitochondria amplified after IVM. Furthermore, it needs to be mentioned that Buravkov et al (2014) also demonstrated that the dye with the advantage to be stable after fixation not only binds sulphhydryl groups in mitochondria, but also in the cytoplasm of cells. Interestingly, it is reported that the content of free cytoplasmic sulphhydryl groups, particularly of glutathione, increases upon oocyte maturation in different species which is dependent on the presence of cumulus cells and precursor molecules (bovine: de Matos, Furnus, & Moses, 1997; horse: Luciano, Goudet, Perazzoli, Lahuec, & Gérard, 2006).…”

Section: Freitasmentioning

confidence: 99%

“…Because of the negative membrane potential, mitochondria accumulate lipophilic cations and several fluorescent probes have been developed to monitor mitochondrial function in living cells (Buravkov, Pogodina, & Buravkova, 2014;Cottet-Rousselle, Ronot, Leverve, & Mayol, 2011;Johnson, Walsh, Bockus, & Chen, 1981;Niżański, Partyka, & Rijsselaere, 2012;Smiley et al, 1991). However, fluorescence emitted by these conventional mitochondrial markers is often susceptible to photobleaching during microscopic examination and the marker exhibit photoinduced toxicity (Chen, 1989).…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, this dye contains a thiol‐reactive group that allows for the preparation of stable specimens by fixation (Poot et al., ). However, recent findings on covalent MitoTracker ® Red CMXRos binding to sulphhydryl groups of proteins and lipids (Buravkov et al., ) require a careful interpretation of fluorescence intensity data in terms of mitochondrial potential. Mitochondrial properties in immature and mature feline oocytes have not been thoroughly studied.…”

Section: Introductionmentioning

confidence: 99%

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Mitochondrial characteristics in oocytes of the domestic cat (Felis catus) after in vitro maturation and vitrification

Sowińska

Müller

Niżański

et al. 2017

Reprod Domestic Animals

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The objective of this study was to evaluate mitochondria in immature and in vitro-matured domestic cat oocytes and to assess for the first time the effect of vitrification on mitochondrial traits. Mitochondrial distribution and aggregation were assessed using confocal microscopy after staining with the fluorescent dye-MitoTracker Red CMXRos. Only cells at the germinal vesicle and the metaphase II stages of nuclear development, representing immature and mature oocytes, respectively, were included in our study. Our study shows that 80% of immature and 100% of mature oocytes exhibit a peripheral pattern of mitochondria distribution, indicating that, in contrast to the situation in other species, the mitochondria of cat oocytes are not dispersed throughout the cell after in vitro maturation but instead maintain a strong affinity for the oocyte periphery near the membrane. However, a loss of aggregation was observed during in vitro maturation-78% of immature oocytes showed homogeneous granulation versus only 18% of mature oocytes (p < .001). The increased intensity of MitoTracker Red CMXRos staining after in vitro maturation (p < .05) may be tentatively attributed to an increase in mitochondrial activity but could likewise reflect a concomitant appearance of sulphhydryl groups in cytoplasm (known to be targeted by the dye). Mitochondrial distribution did not change upon vitrification; however, dye intensity decreased (p < .05) and mitochondrial aggregation was intensified in both immature and mature vitrified cat oocytes.

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Comparison of Mitochondrial Fluorescent Dyes in Stromal Cells

Cited by 11 publications

References 11 publications

Targeting eIF5A Hypusination Prevents Anoxic Cell Death through Mitochondrial Silencing and Improves Kidney Transplant Outcome

Targeting eIF5A Hypusination Prevents Anoxic Cell Death through Mitochondrial Silencing and Improves Kidney Transplant Outcome

Mitochondrial-Localized Versus Cytosolic Intracellular CO-Releasing Organic PhotoCORMs: Evaluation of CO Effects Using Bioenergetics

Mitochondrial characteristics in oocytes of the domestic cat (Felis catus) after in vitro maturation and vitrification

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