The effect of a typical magnetic storm on mitosis in the embryo cells and the length and weight of roach (Rutilus rutilus L.) prolarvae

The action of the tested variants of a typical magnetic storm (MS) has a biological effect on cellular and organismal levels in the prolarva stage. The impact of this environmental factor leads to increased mitotic activity in blastocysts, it can trigger the early hatching of embryos, and it has no negative impact on the size-weight characteristics of free embryos.     

Some morphological features of fry of roach <Emphasis Type="Italic">Rutilus rutilus</Emphasis> (Cyprinidae,Cypriniformes) after separate and combined exposure of embryos to magnetic field and elevated temperature

Separate and combined exposure of roach Rutilus rutilus embryos during the first 38 h after fertilization to magnetic field (500 Hz, 1.4–1.6 µT) and elevated temperature (23°?) causes morphological responses: decrease in mean values of length and weight of the body, increase in the number of vertebrae, and increase in the number of vertebral anomalies and morphological diversity of vertebral phenotypes. Size–weight characteristics and the number of vertebrae in underyearlings are related to the development of anomalies in its backbone. A method for counting of vertebrae in fish with backbone malformations is suggested.     

Involvement of chloroplasts in the programmed death of plant cells

The effect of cyanide, an apoptosis inducer, on pea leaf epidermal peels was investigated. Illumination stimulated the CN^–-induced destruction of guard cells (containing chloroplasts and mitochondria) but not of epidermal cells (containing mitochondria only). The process was prevented by antioxidants (-tocopherol, 2,5-di-tret-butyl-4-hydroxytoluene, and mannitol), by anaerobiosis, by the protein kinase C inhibitor staurosporine, and by cysteine and serine protease inhibitors. Electron acceptors (menadione, p-benzoquinone, diaminodurene, TMPD, DCPIP, and methyl viologen) suppressed CN^–-induced apoptosis of guard cells, but not epidermal cells. Methyl viologen had no influence on the removal of CN^–-induced nucleus destruction in guard cells under anaerobic conditions. The light activation of CN^–-induced apoptosis of guard cells was suppressed by DCMU (an inhibitor of the electron transfer in Photosystem II) and by DNP-INT (an antagonist of plastoquinol at the Q_o site of the chloroplast cytochrome b ₆ f complex). It is concluded that apoptosis initiation in guard cells depends on the simultaneous availability of two factors, ROS and reduced quinones of the electron transfer chain. The conditions for manifestation of programmed cell death in guard and epidermal cells of the pea leaf were significantly different.     

Inhibition of mitochondrial bioenergetics: the effects on structure of mitochondria in the cell and on apoptosis

The effects of specific inhibitors of respiratory chain, F(o)F(1)ATP synthase and uncouplers of oxidative phosphorylation on survival of carcinoma HeLa cells and on the structure of mitochondria in the cells were studied. The inhibitors of respiration (piericidin, antimycin, myxothiazol), the F(1)-component of ATP synthase (aurovertin) and uncouplers (DNP, FCCP) did not affect viability of HeLa cells, apoptosis induced by TNF or staurosporin and the anti-apoptotic action of Bcl-2. Apoptosis was induced by combined action of respiratory inhibitors and uncouplers indicating possible pro-apoptotic action of reactive oxygen species (ROS) generated by mitochondria. Short-term incubation of HeLa cells with the mitochondrial inhibitors and 2-deoxyglucose followed by 24-48 h recovery resulted in massive apoptosis. Apoptosis correlated to transient (3-4 h) and limited (60-70%) depletion of ATP. More prolonged or more complete transient ATP depletion induced pronounced necrosis. The inhibitors of respiration and uncouplers caused fragmentation of tubular mitochondria and formation of small round bodies followed by swelling. These transitions were not accompanied with release of cytochrome c into the cytosol and were fully reversible. The combined effect of respiratory inhibitors and uncouplers developed more rapidly indicating possible involvement of ROS generated by mitochondria. More prolonged (48-72 h) incubation with this combination of inhibitors caused clustering and degradation of mitochondria.     

Novel mitochondria-targeted compounds composed of natural constituents: Conjugates of plant alkaloids berberine and palmatine with plastoquinone

Novel mitochondria-targeted compounds composed entirely of natural constituents have been synthesized and tested in model lipid membranes, in isolated mitochondria, and in living human cells in culture. Berberine and palmatine, penetrating cations of plant origin, were conjugated by nonyloxycarbonylmethyl residue with the plant electron carrier and antioxidant plastoquinone. These conjugates (SkQBerb, SkQPalm) and their analogs lacking the plastoquinol moiety (C10Berb and C10Palm) penetrated across planar bilayer phospholipid membrane in their cationic forms and accumulated in isolated mitochondria or in mitochondria in living human cells in culture. Reduced forms of SkQBerb and SkQPalm inhibited lipid peroxidation in isolated mitochondria at nanomolar concentrations. In isolated mitochondria and in living cells, the berberine and palmatine moieties were not reduced, so antioxidant activity belonged exclusively to the plastoquinol moiety. In human fibroblasts, nanomolar SkQBerb and SkQPalm prevented fragmentation of mitochondria and apoptosis induced by exogenous hydrogen peroxide. At higher concentrations, conjugates of berberine and palmatine induced proton transport mediated by free fatty acids both in model and in mitochondrial membrane. In mitochondria this process was facilitated by the adenine nucleotide carrier. As an example of application of the novel mitochondria-targeted antioxidants SkQBerb and SkQPalm to studies of signal transduction, we discuss induction of cell cycle arrest, differentiation, and morphological normalization of some tumor cells. We suggest that production of oxygen radicals in mitochondria is necessary for growth factors-MAP-kinase signaling, which supports proliferation and transformed phenotype.     

Some morphological features of roach <Emphasis Type="Italic">Rutilus rutilus</Emphasis> (Cyprindae) fry after exposure to toxicants in the early stages of ontogenesis (vertebral phenotypes,plastic features,and fluctuating assymetry)

Roach Rutilus rutilus eggs developed in low-concentration solutions of trichlorfon (10^?6, 10^?5, 10^?4, 10^?3, and 10^?2 mg/l) or N-methyl-N’-nitro-N-nitrosoguanidin (MNNG) with 0.3 mg/l concentration for 54 h after fertilization. Roach fry from different experimental variants differed from the control in terms of growth rate and number of vertebrae in the spinal column and its sections. Irrespective of a change in the direction of growth rate, the number of vertebrae in the spinal column and diversity of vertebral phenotypes increased. The number of vertebrae and Shannon’s index of vertebral column phenotype diversity correlated with the number of anomalies in the structure of axial skeleton. Body proportions of fish with anomalies change due to shortening of deformed fragments of the vertebral column: relative indices of head length and maximum body depth increase. Regarding dispersion of fluctuating asymmetry for the number of openings of seismosensory system in the frontale, dentale and praeoperculum, insignificant differences from the control were found in two cases only: it was lower for the number of openings in dentale in fry exposed to trichlorfon with a concentration of 10^?5 mg/l, and higher for the number of openings in praeoperculum in the variant with MNNG.     

Production of reactive oxygen species in mitochondria of HeLa cells under oxidative stress

Mitochondria can be a source of reactive oxygen species (ROS) and a target of oxidative damage during oxidative stress. In this connection, the effect of photodynamic treatment (PDT) with Mitotracker Red (MR) as a mitochondria-targeted photosensitizer has been studied in HeLa cells. It is shown that MR produces both singlet oxygen and superoxide anion upon photoactivation and causes photoinactivation of gramicidin channels in a model system (planar lipid bilayer). Mitochondria-targeted antioxidant (MitoQ) inhibits this effect. In living cells, MR-mediated PDT initiates a delayed ("dark") accumulation of ROS, which is accelerated by inhibitors of the respiratory chain (piericidin, rotenone and myxothiazol) and inhibited by MitoQ and diphenyleneiodonium (an inhibitor of flavin enzymes), indicating that flavin of Complex I is involved in the ROS production. PDT causes necrosis that is prevented by MitoQ. Treatment of the cell with hydrogen peroxide causes accumulation of ROS, and the effects of inhibitors and MitoQ are similar to that described for the PDT model. Apoptosis caused by H2O2 is augmented by the inhibitors of respiration and suppressed by MitoQ. It is concluded that the initial segments of the respiratory chain can be an important source of ROS, which are targeted to mitochondria, determining the fate of the cell subjected to oxidative stress.     

Behavioral and morphological asymmetries in roach <Emphasis Type="Italic">Rutilus rutilus</Emphasis> (Cyprinidae: Cypriniformes) underyearlings

The underyearlings of roach, Rutilus rutilus, displayed a bilateral asymmetry of C-bend—the initial stage of escape behavior. Most individuals exhibited a significant bias to turn leftward or rightward after being stimulated by electrical current. This individual asymmetry was consistent when the same fish were retested ten days later. A significant correlation was revealed between the behavioral asymmetry and the bilateral asymmetry of surface area of frontal (positive correlation) and parietal (negative correlation) cranial bones. No significant correlation was found with the bilateral asymmetry of other morphological characteristics: numbers of pores of the seismosensory canals (praeopercular-mandibular, supraorbital, and supratemporal) in flat cranial bones (dental, praeopercular, frontal, and parietal numbers of lateral line pores, and numbers of rays in pectoral and ventral fins.     

"Wages of fear": transient threefold decrease in intracellular ATP level imposes apoptosis

In HeLa cells, complete inhibition of oxidative phosphorylation by oligomycin, myxothiazol or FCCP combined with partial inhibition of glycolysis by DOG resulted in a steady threefold decrease in the intracellular ATP level. The ATP level recovers when the DOG-containing medium was replaced by that with high glucose. In 48 h after a transient (3 h) [ATP] lowering followed by recovery of the ATP level, the majority of the cells commits suicide by means of apoptosis. The cell death does not occur if DOG or an oxidative phosphorylation inhibitor was added separately, treatments resulting in 10-35% lowering of [ATP]. Apoptosis is accompanied by Bax translocation to mitochondria, cytochrome c release into cytosol, caspase activation, reactive oxygen species (ROS) generation, and reorganization and decomposition of chromatin. Apoptosis appears to be sensitive to oncoprotein Bcl-2 and a pancaspase inhibitor zVADfmk. In the latter case, necrosis is shown to develop instead of apoptosis. The cell suicide is resistant to cyclosporine A, a phospholipase inhibitor trifluoroperazine, the JNK and p38 kinase inhibitors, oligomycin, N-acetyl cysteine and mitoQ, differing in these respects from the tumor necrosis factor (TNF)- and H(2)O(2)-induced apoptoses. It is suggested that the ATP concentration in the cell is monitored by intracellular "ATP-meter(s)" generating a cell suicide signal when ATP decreases, even temporarily, below some critical level (around 1 mM).