[2] Long-term storage of mitochondria to preserve energy-linked functions

Fleischer, Sidney

doi:10.1016/0076-6879(79)55004-6

Cited by 15 publications

(6 citation statements)

References 7 publications

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“…Bars in (B) and (C) represent group means (S.E.M.). 1979;Fuller et al, 1989;De Loecker et al, 1991;Kuznetsov et al, 2003). Cryopreservation was also successfully applied to primary rat hippocampal neurons (Mattson and Kater, 1988), rat brain synaptosomes (Begley et al, 1998), rat cardiac and skeletal muscles and human biopsy samples (Kuznetsov et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation of brain mitochondria: A novel methodology for functional studies

Nukala

Singh

Davis

et al. 2006

Journal of Neuroscience Methods

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Section: Discussionmentioning

confidence: 99%

Cryopreservation of brain mitochondria: A novel methodology for functional studies

Nukala

Singh

Davis

et al. 2006

Journal of Neuroscience Methods

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“…Specifically, mitochondria were purified by differential centrifugation, with 2-3 additional washing steps implemented for experiments in Figure 5, Figure 6D, Supplemental Figure 4A, Supplemental Figure 4, C-F, and Supplemental Figure 5; by density gradient centrifugation for experiments in Figure 2F, Figure 3B, Figure 4, Figure 6, A-C, Figure 7, A-C, Figure 8, and Supplemental Figure 3; by a combination of density gradient centrifugation and ZE-FFE purification for experiments in Figure 2, A-E, Figure 3A, Figure 3C, Figure 7D, and Supplemental Figure 2, B-E. Fresh mitochondria were used for respiratory measurements or copper and calcium challenge experiments and were sub-sequently analyzed for swelling, transmembrane potential (Δψm), and ATP synthesis or subjected to ZE-FFE analysis, electron microscopy, and redox proteomics. Mitochondrial samples isolated by density gradient purification were stored at -192°C in preservation buffer (50) and used for the assessments of enzymatic activities, lipid peroxidation, and quantification of structural alterations. For the latter analysis, samples were subsequently purified by ZE-FFE on the same day and subjected to electron microscopy analysis in parallel.…”

Section: Methodsmentioning

confidence: 99%

Liver mitochondrial membrane crosslinking and destruction in a rat model of Wilson disease

Zischka

Lichtmannegger²,

Schmitt³

et al. 2011

J. Clin. Invest.

161

162

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Wilson disease (WD) is a rare hereditary condition that is caused by a genetic defect in the copper-transportingATPase ATP7B that results in hepatic copper accumulation and lethal liver failure. The present study focuses on the structural mitochondrial alterations that precede clinical symptoms in the livers of rats lacking Atp7b, an animal model for WD. Liver mitochondria from these Atp7b -/-rats contained enlarged cristae and widened intermembrane spaces, which coincided with a massive mitochondrial accumulation of copper. These changes, however, preceded detectable deficits in oxidative phosphorylation and biochemical signs of oxidative damage, suggesting that the ultrastructural modifications were not the result of oxidative stress imposed by copper-dependent Fenton chemistry. In a cell-free system containing a reducing dithiol agent, isolated mitochondria exposed to copper underwent modifications that were closely related to those observed in vivo. In this cell-free system, copper induced thiol modifications of three abundant mitochondrial membrane proteins, and this correlated with reversible intramitochondrial membrane crosslinking, which was also observed in liver mitochondria from Atp7b -/-rats. In vivo, copper-chelating agents reversed mitochondrial accumulation of copper, as well as signs of intra-mitochondrial membrane crosslinking, thereby preserving the functional and structural integrity of mitochondria. Together, these findings suggest that the mitochondrion constitutes a pivotal target of copper in WD.

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“…Yeast and rat liver mitochondria were extracted from cells and organs by standard methods (4,6,27). S. cerevisiae was streaked onto a YPD agar plate and precultured at 30°C overnight.…”

Section: Animalsmentioning

confidence: 99%

T-2307 Causes Collapse of Mitochondrial Membrane Potential in Yeast

Shibata

Takahashi

Yamada

et al. 2012

Antimicrob Agents Chemother

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T-2307, an arylamidine compound, has been previously reported to have broad-spectrum in vitro and in vivo antifungal activities against clinically significant pathogens, including Candida species, Cryptococcus neoformans, and Aspergillus species, and is now undergoing clinical trials. Here we investigated the mechanism of action of T-2307 using yeast cells and mitochondria isolated from yeast and rat liver. Nonfermentative growth of Candida albicans and Saccharomyces cerevisiae in glycerol medium, in which yeasts relied on mitochondrial respiratory function, was inhibited at 0.001 to 0.002 g/ml (0.002 to 0.004 M) of T-2307. However, fermentative growth in dextrose medium was not inhibited by T-2307. Microscopic examination using Mitotracker fluorescent dye, a cell-permeant mitochondrion-specific probe, demonstrated that T-2307 impaired the mitochondrial function of C. albicans and S. cerevisiae at concentrations near the MIC in glycerol medium. T-2307 collapsed the mitochondrial membrane potential in mitochondria isolated from S. cerevisiae at 20 M. On the other hand, in isolated rat liver mitochondria, T-2307 did not have any effect on the mitochondrial membrane potential at 10 mM. Moreover, T-2307 had little inhibitory and stimulatory effect on mitochondrial respiration in rat liver mitochondria. In conclusion, T-2307 selectively disrupted yeast mitochondrial function, and it was also demonstrated that the fungal mitochondrion is an attractive antifungal target.

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[2] Long-term storage of mitochondria to preserve energy-linked functions

Cited by 15 publications

References 7 publications

Cryopreservation of brain mitochondria: A novel methodology for functional studies

Cryopreservation of brain mitochondria: A novel methodology for functional studies

Liver mitochondrial membrane crosslinking and destruction in a rat model of Wilson disease

T-2307 Causes Collapse of Mitochondrial Membrane Potential in Yeast

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