Biomedical subjects

R Morkuniene

Publications and source records attributed to R Morkuniene.

7 recordsLinked to original sources

PubMed2001

Release of mitochondrial cytochrome c and activation of cytosolic caspases induced by myocardial ischaemia.

It has previously been shown that apoptosis is increased in ischaemic/reperfused heart. However, little is known about the mechanism of induction of apoptosis in myocardium during ischaemia. We investigated whether prolonged myocardial ischaemia causes activation of caspases and whether this activation is related to cytochrome c release from mitochondria to cytosol during ischaemia. Using an in vitro model of heart ischaemia, we show that 60 min ischaemia leads to a significant accumulation of cytochrome c in the cytosol and a decrease in mitochondrial content of cytochrome c but not cytochrome a. The release of cytochrome c from mitochondria was accompanied by activation of caspase-3-like proteases (measured by cleavage of fluorogenic peptide substrate DEVD-amc) and a large increase in number of cells with DNA strand breaks (measured by TUNEL staining). Caspase-1-like proteases (measured by YVAD-amc cleavage) were not activated during ischaemia. Addition of 14 microM cytochrome c to cytosolic extracts prepared from control hearts induced ATP-dependent activation of caspase-3-like protease activity. Our data suggest that extended heart ischaemia can cause apoptosis mediated by release of cytochrome c from mitochondria and subsequent activation of caspase-3.

Animals↗

PubMed2001

137Cs and 90Sr behavioural regularities in the southeastern part of the Baltic Sea.

Variations in 137Cs concentrations were investigated over the period 1986-1997 in the southeastern part of the Baltic Sea, following the Chernobyl power plant accident. The rate of "self-cleaning" was demonstrated to be very slow, the average concentration of 137Cs in 1996 being almost the same as that measured directly after the accident, in 1986. Measurements of both 137Cs and 90Sr concentrations generally revealed homogeneous distributions in this region of the Baltic Sea, though patchy distributions did develop under some hydrometeorological conditions. Specifically, the 137Cs concentration distribution became heterogeneous with values varying in the range 60-92 Bq/m3 under south-southwesterly wind conditions whilst the 90Sr concentration distribution developed similar characteristics with values ranging from 15 to 64 Bq/m3 under east-southeasterly wind conditions. In addition, in coastal waters, over extensive periods of north-northwesterly winds in 1995, 137Cs concentrations increased to values 1.5-2 times the overall average concentration, which was registered in 1986 and 1996. These data therefore reveal a continuing significant pollution of the waters of the Baltic Sea resulting from the Chernobyl power plant accident, a pollution compounded by the slow rate of radionuclide self-cleaning and significant probability of sudden regional concentration increase.

Baltic States↗

PubMed2000

Nitric oxide donors, nitrosothiols and mitochondrial respiration inhibitors induce caspase activation by different mechanisms.

We investigated to what extent different types of NO donors induce caspase activation by opening of the mitochondrial permeability transition pore (PTP) or inhibition of mitochondrial respiration. We found that nitrosothiols can directly open the PTP in isolated mitochondria and cause cytochrome c release, whereas NONOate donors can not. In macrophages nitrosothiols cause caspase activation that is blocked by cyclosporin A or calcium chelation, both of which prevent PTP opening, whereas caspase activation caused by NONOates is much less sensitive to these agents. Inhibitors of mitochondrial respiration did not promote PTP opening in isolated mitochondria, and although they cause caspase activation in macrophages, this activation was slower than that caused by NO donors, and was relatively insensitive to cyclosporin and calcium chelators suggesting that PTP opening was not involved.

Animals↗

PubMed1999

Release of cytochrome c from heart mitochondria is induced by high Ca2+ and peroxynitrite and is responsible for Ca(2+)-induced inhibition of substrate oxidation.

Prolonged heart ischaemia causes an inhibition of oxidative phosphorylation and an increase of Ca2+ in mitochondria. We investigated whether elevated Ca2+ induces changes in the oxidative phosphorylation system relevant to ischaemic damage, and whether Ca2+ and other inducers of mitochondrial permeability transition cause the release of cytochrome c from isolated heart mitochondria. We found that 5 microM free Ca2+ induced changes in oxidative phosphorylation system similar to ischaemic damage: increase in the proton leak and inhibition of the substrate oxidation system related to the release of cytochrome c from mitochondria. The phosphorylating system was not directly affected by high Ca2+ and ischaemia. The release of cytochrome c from mitochondria was caused by Ca2+ and 0.175-0.9 mM peroxynitrite but not by NO, and was prevented by cyclosporin A. Adenylate kinase and creatine kinase were also released after incubation of mitochondria with Ca2+, however, the activity of citrate synthase in the incubation medium with high and low Ca2+ did not change. The data suggest that release of cytochrome c and other proteins of intermembrane space may be due to the opening of the mitochondrial permeability transition pore, and may be partially responsible for inhibition of mitochondrial respiration induced by ischaemia, high calcium, and oxidants.

Animals↗

PubMed1997

Metabolic control analysis and threshold effect in oxidative phosphorylation: implications for mitochondrial pathologies.

We have shown that the Metabolic Control Analysis (MCA) can explain the threshold effect observed in the expression of mitochondrial diseases. As a matter of fact, the effect of a specific inhibitor on the flux of O2 consumption mimics a defect in a step of oxidative phosphorylation. The observed threshold is correlated to the value of the control coefficient of the inhibited step. For this reason, we have studied the repartition of the control coefficients of different steps in oxidative phosphorylation on various tissues (liver, kidney, brain, skeletal muscle and heart). We discuss the results in terms of metabolic control theory and provide a possible explanation for the heterogeneous phenotype of those pathologies. We present the double threshold hypothesis of both a threshold in the energy demand of a tissue and in the energy supply by oxidative phosphorylation.

Animals↗

PubMed1996

Ca2+ stimulates both the respiratory and phosphorylation subsystems in rat heart mitochondria.

Stimulation of mitochondrial respiration by physiological concentrations of Ca2+ was studied to determine which components of oxidative phosphorylation are affected by Ca2+. The kinetic dependence of the respiratory chain, phosphorylation subsystem and proton leak on the mitochondrial membrane potential in isolated rat heart mitochondria respiring on 2-oxoglutarate or succinate was measured at two different concentrations of external free Ca2+. The results show that proton leak is not directly affected by Ca2+, but that both the respiratory and phosphorylation systems can be directly stimulated by Ca2+ depending on conditions. Although Ca2+ directly stimulates the phosphorylation system, this has relatively little effect on respiration rate with 2-oxoglutarate in States 3 and 4 because the subsystem has little control over respiration. However, in intermediate states, the phosphorylation system has greater control and Ca2+ stimulation of this system contributes substantially to the stimulation of respiration and phosphorylation. In the case of succinate oxidation neither the respiratory subsystem nor the phosphorylation system is stimulated by Ca2+.

Animals↗

PubMed1996

Kinetic analysis of changes in activity of heart mitochondrial oxidative phosphorylation system induced by ischemia.

The authors have previously shown that ischemia causes inhibition of the respiratory chain and phosphorylation system, and stimulation of the proton leak of mitochondria isolated from rat heart. It is shown here that the activity of the mitochondrial respiratory chain (after 30 min ischemia, but not after 45 min) and the phosphorylation system are completely restored to the normal level by the addition of exogenous cytochrome c when succinate is used as substrate. Moreover, cytochrome c causes apparent activation of the respiratory chain, the phosphorylation system and the proton leak in normal mitochondria. This can be explained by a fraction of the mitochondrial population lacking cytochrome c and this fraction may increase with ischemia. Experiments on skinned cardiac fibers showed that cytochrome c has no effect on mitochondrial respiration after 15 min ischemia, but the stimulation of respiration by cytochrome c progressively increases when ischemia was prolonged up to 30 min and 45 min, suggesting that the loss of cytochrome c may occur in vivo during the early reversible phase of ischemia. Mitochondria isolated from hearts after 45 min ischemia have a defect in the respiratory chain unrelated to the loss of cytochrome c. These mitochondria have an increased level of Ca2+: 10.05 nmol/mg protein compared to 4.64 nmol/mg in control mitochondria and 4.32 nmol/mg in 30 min ischemic mitochondria. The increase in the proton leak in ischemic mitochondria is mostly reversible by albumin and thus, may be related to an increased level of free fatty acids in ischemic mitochondria.

Animals↗