Improved methods to isolate and subfractionate rat liver mitochondria. Lipid composition of the inner and outer membrane

Hovius, Ruud; Lambrechts, Hilde; Nicolay, Klaas; Kruijff, Ben de

doi:10.1016/0005-2736(90)90036-n

Cited by 340 publications

(227 citation statements)

References 47 publications

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“…Numerous attempts to diminish microsomal contamination, using multiple washes, centrifugation over a sucrose cushion or sucrose gradient [8] and osmotic swelling followed by shrinkage [37,38] were unsuccessful. Addition of a re-homogenization step prior to osmotic shock, however, reduced microsomal contamination to 27% suggesting that the microsomes were closely attached to mitochondrial fragments but could be dislodged by physical disruption.…”

Section: Resultsmentioning

confidence: 99%

Mitochondrial P450-dependent arachidonic acid metabolism by TCDD-induced hepatic CYP1A5; conversion of EETs to DHETs by mitochondrial soluble epoxide hydrolase

Labitzke¹,

Diani-Moore²,

Rifkind³

2007

Archives of Biochemistry and Biophysics

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Several P450 enzymes localized in the endoplasmic reticulum and thought to be involved primarily in xenobiotic metabolism, including mouse and rat CYP1A1 and mouse CYP1A2, have also been found to translocate to mitochondria. We report here that the environmental toxin 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces enzymatically active CYP1A4/1A5, the avian orthologs of mammalian CYP1A1/1A2, in chick embryo liver mitochondria as well as in microsomes. P450 proteins and activity levels (CYP1A4-dependent 7-ethoxyresorufin-O-deethylase and CYP1A5-dependent arachidonic acid epoxygenation) in mitochondria were 23-40% of those in microsomes. DHET formation by mitochondria was twice that of microsomes and was attributable to a mitochondrial soluble epoxide hydrolase as confirmed by Western blotting with antiEPHX2, conversion by mitochondria of pure 11,12 and 14,15-EET to the corresponding DHETs and inhibition of DHET formation by the soluble epoxide hydrolase inhibitor, 12(-3-adamantan-1-yl-ureido)-dodecanoic acid (AUDA). TCDD also suppressed formation of mitochondrial and microsomal 20-HETE. The findings newly identify mitochondria as a site of P450-dependent arachidonic acid metabolism and as a potential target for TCDD effects. They also demonstrate that mitochondria contain soluble epoxide hydrolase and underscore a role for CYP1A in endobiotic metabolism.

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Section: Resultsmentioning

confidence: 99%

Mitochondrial P450-dependent arachidonic acid metabolism by TCDD-induced hepatic CYP1A5; conversion of EETs to DHETs by mitochondrial soluble epoxide hydrolase

Labitzke¹,

Diani-Moore²,

Rifkind³

2007

Archives of Biochemistry and Biophysics

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“…36 The mouse brain was washed in 300 mM sucrose, 0.1 mM EGTA, 10 mM HEPES, pH 7.4 (buffer A), homogenized in 15 ml of buffer A using a motor-driven glass Potter-Elvehjem homogenizer with PTFE pestle. The cell debris and nuclei were centrifuged at 600 Â g for 5 min, and the supernatant was filtered through 4 layers of cheesecloth.…”

Section: Subfractionation Of Mouse Brain Mitochondria By the Swellingmentioning

confidence: 99%

EndoG is dispensable in embryogenesis and apoptosis

Sasaki

et al. 2005

Cell Death Differ

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The mitochondrial protein, endonuclease G (EndoG), is one of the endonucleases implicated in DNA fragmentation during apoptosis. It has been shown to translocate from the mitochondria to the nucleus upon cell death stimuli. These observations suggest that EndoG is a mitochondrial cell death effector, and that it possibly acts as a cell death nuclease, similar to DNA fragmentation factor. To better understand the role of EndoG in development and apoptosis, we generated EndoG null mice by homologous gene targeting without disruption of D2Wsu81e. EndoG null mice are viable and develop to adulthood with no obvious abnormalities. Fibroblasts generated from the EndoG null mice show no difference in susceptibility when induced to die by a variety of intrinsic and extrinsic apoptotic stimuli. Additionally, EndoG null mice are equally sensitive to excitotoxic stress. These data suggest that EndoG is not essential for early embryogenesis and apoptosis.

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“…Mitochondria were prepared by differential centrifugation followed by Percoll purification [27]. Briefly, striata from four mice were excised, weighed, washed with 0.25 M sucrose, and homogenized (1/8, w/v) in a medium containing 250 mM mannitol, 0.5 mM EGTA, 0.1% (w/v) BSA, 5 mM Hepes/KOH, pH 7.4 at 4 • C. The homogenate was centrifuged for 5 min at 600 × g and the supernatant was centrifuged at 10,300 × g for 10 min.…”

Section: Complex I (Nadh: Ubiquinone Oxidoreductase) Assaymentioning

confidence: 99%

Synergistic effects of melatonin and deprenyl against MPTP-induced mitochondrial damage and DA depletion

Khaldy

Escames

León

et al. 2003

Neurobiology of Aging

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Previous studies showed a synergistic effect of melatonin and deprenyl against dopamine (DA) autoxidation in vitro. Since oxidative stress is implicated in Parkinson's disease (PD), we explored the effects of melatonin plus deprenyl administration in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD in C57/Bl6 mice. Melatonin, but not deprenyl prevents the inhibition of mitochondrial complex I and the oxidative damage in nigrostriatal neurons induced by MPTP. With the dose used deprenyl recovers 50% DA levels and tyrosine hydroxylase activity depressed by the neurotoxin, normalizing locomotor activity of mice. Melatonin, which was unable to counteract MPTP-induced DA depletion and inhibition of tyrosine hydroxylase activity, potentiates the effect of deprenyl on catecholamine turnover and mice ambulatory activity. These results suggest a dissociation of complex I inhibition from DA depletion in this model of Parkinson's disease. The data also support that a combination of melatonin, which improves mitochondrial electron transport chain and reduces oxidative damage, and deprenyl, which promotes the specific function of the rescued neurons, i.e. DA turnover, may be a promising strategy for the treatment of PD.

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Improved methods to isolate and subfractionate rat liver mitochondria. Lipid composition of the inner and outer membrane

Cited by 340 publications

References 47 publications

Mitochondrial P450-dependent arachidonic acid metabolism by TCDD-induced hepatic CYP1A5; conversion of EETs to DHETs by mitochondrial soluble epoxide hydrolase

Mitochondrial P450-dependent arachidonic acid metabolism by TCDD-induced hepatic CYP1A5; conversion of EETs to DHETs by mitochondrial soluble epoxide hydrolase

EndoG is dispensable in embryogenesis and apoptosis

Synergistic effects of melatonin and deprenyl against MPTP-induced mitochondrial damage and DA depletion

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