Epilepsy is a neurologic disorder, particularly frequent in infants and children where it can lead to serious consequences later in life. Oxidative stress and mitochondrial dysfunction are implicated in the pathogenesis of many neurological disorders including epilepsy in adults. However, their role in immature epileptic brain is unclear since there have been two contrary opinions: oxidative stress is age-dependent and does not occur in immature brain during status epilepticus (SE) and, on the other hand, evidence of oxidative stress in immature brain during a specific model of SE. To solve this dilemma, we have decided to investigate oxidative stress following SE induced in immature 12-day-old rats by three substances with a different mechanism of action, namely 4-aminopyridine, LiCl-pilocarpine or kainic acid. Fluoro-Jade-B staining revealed mild brain damage especially in hippocampus and thalamus in each of the tested models. Decrease of glucose and glycogen with parallel rises of lactate clearly indicate high rate of glycolysis, which was apparently not sufficient in 4-AP and Li-Pilo status, as evident from the decreases of PCr levels. Hydroethidium method revealed significantly higher levels of superoxide anion (by ∼60%) in the hippocampus, cerebral cortex and thalamus of immature rats during status. SE lead to mitochondrial dysfunction with a specific pronounced decrease of complex I activity that persisted for a long period of survival. Complexes II and IV activities remained in the control range. Antioxidant treatment with SOD mimetic MnTMPYP or peroxynitrite scavenger FeTPPS significantly attenuated oxidative stress and inhibition of complex I activity. These findings bring evidence that oxidative stress and mitochondrial dysfunction are age and model independent, and may thus be considered a general phenomenon. They can have a clinical relevance for a novel approach to the treatment of epilepsy, allowing to target the mechanisms which play a crucial or additive role in the pathogenesis of epilepsies in infants and children.

Institute of Physiology The Czech Academy of Sciences Prague Czech Republic

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Bindokas V. P., Jordán J., Lee C. C., Miller R. J. (1996). Superoxide production in rat hippocampal neurons: selective imaging with hydroethidine. PubMed PMC

Bruce A. J., Baudry M. (1995). Oxygen free radicals in rat limbic structures after kainate-induced seizures. PubMed

Dobbing J. (1970). “Undernutrition and the developing brain,” in

Fato R., Bergamini C., Leoni S., Strocchi P., Lenaz G. (2008). Generation of reactive oxygen species by mitochondrial complex I: implications in neurodegeneration. PubMed DOI

Folbergrová J. (1993). Cerebral energy state of neonatal rats during seizures induced by homocysteine. PubMed

Folbergrová J. (1997). Anticonvulsant action of both NMDA and non-NMDA receptor antagonists against seizures induced by homocysteine in immature rats. PubMed DOI

Folbergrová J. (2013). Oxidative stress in immature brain following experimentally-induced seizures. PubMed

Folbergrová J., Druga R., Haugvicová R., Mareš P., Otáhal J. (2008). Anticonvulsant and neuroprotective effect of (S)-3,4-dicarboxyphenylglycine against seizures induced in immature rats by homocysteic acid. PubMed DOI

Folbergrová J., Druga R., Otáhal J., Haugvicová R., Mareš P., Kubová H. (2005). Seizures induced in immature rats by homocysteic acid and the associated brain damage are prevented by group II metabotropic glutamate receptor agonist (2R,4R)-4-aminopyrrolidine-2,4-dicarboxylate. PubMed DOI

Folbergrová J., Druga R., Otáhal J., Haugvicová R., Mareš P., Kubová H. (2006). Effect of free radical spin trap N-tert-butyl-α-phenylnitrone (PBN) on seizures induced in immature rats by homocysteic acid. PubMed DOI

Folbergrová J., Haugvicová R., Mareš P. (2000). Behavioral and metabolic changes in immature rats during seizures induced by homocysteic acid: the protective effect of NMDA and non-NMDA receptor antagonists. PubMed DOI

Folbergrová J., Ješina P., Drahota Z., Lisý V., Haugvicová R., Vojtíšková A., et al. (2007). Mitochondrial complex I inhibition in cerebral cortex of immature rats following homocysteic acid-induced seizures. PubMed DOI

Folbergrová J., Ješina P., Haugvicová R., Lisý V., Houštěk J. (2010). Sustained deficiency of mitochondrial complex I activity during long periods of survival after seizures induced in immature rats by homocysteic acid. PubMed DOI

Folbergrová J., Kunz W. S. (2012). Mitochondrial dysfunction in epilepsy. PubMed DOI

Folbergrová J., MacMillan V., Siesjö B. K. (1972). The effect of moderate and marked hypercapnia upon the energy state and upon the cytoplasmic NADH/NAD+ ratio of the rat brain. PubMed DOI

Folbergrová J., Otáhal J., Druga R. (2011). Effect of Tempol on brain superoxide anion production and neuronal injury associated with seizures in immature rats. DOI

Folbergrová J., Otáhal J., Druga R. (2012). Brain superoxide anion formation in immature rats during seizures: protection by selected compounds. PubMed DOI

Hirsch E., Baram T. Z., Snead O. C. (1992). Ontogenic study of lithium–pilocarpine-induced status epilepticus in rats. PubMed DOI

Jensen F. E. (1999). Acute and chronic effects of seizures in the developing brain: experimental models. PubMed DOI

Kalyanaraman B., Dranka B. P., Hardy M., Michalski R., Zielonka J. (2014). HPLC-based monitoring of products formed from hydroethidine-based fluorogenic probes-the ultimate approach for intra- and extracellular superoxide detection. PubMed DOI PMC

Kann O., Kovács R., Njunting M., Behrens C. J., Otáhal J., Lehmann T. N., et al. (2005). Metabolic dysfunction during neuronal activation in the ex vivo hippocampus from chronic epileptic rats and humans. PubMed DOI

Kubová H., Druga R., Lukasiuk K., Suchomelová L., Haugvicová R., Jirmanová I., et al. (2001). Status epilepticus causes necrotic damage in the mediodorsal nucleus of the thalamus in immature rats. PubMed PMC

Kubová H., Mareš P., Suchomelová L., Brožek G., Druga R., Pitkänen A. (2004). Status epilepticus in immature rats leads to behavioural and cognitive impairment and epileptogenesis. PubMed DOI

Kudin A. P., Bimpong-Buta N. Y. B., Vielhaber S., Elger C. E., Kunz W. S. (2004). Characterization of superoxide-producing sites in isolated brain mitochondria. PubMed DOI

Kunz W. S., Kudin A. P., Vielhaber S., Blümcke I., Zuschratter W., Schramm J., et al. (2000). Mitochondrial complex I deficiency in the epileptic focus of patients with temporal lobe epilepsy. PubMed DOI

Kussmaul L., Hirst J. (2006). The mechanism of superoxide production by NADH:ubiquinone oxidoreductase (complex I) from bovine heart mitochondria. PubMed DOI PMC

Lee Y. M., Kang H. C., Lee J. S., Kim S. H., Kim E. Y., Lee S. K., et al. (2008). Mitochondrial respiratory chain defects: underlying etiology in various epileptic conditions. PubMed DOI

Liang L. P., Ho Y. S., Patel M. (2000). Mitochondrial superoxide production in kainate-induced hippocampal damage. PubMed DOI

Liang L. P., Patel M. (2004). Mitochondrial oxidative stress and increased seizure susceptibility in SOD2 (-/+) mice. PubMed DOI

Lin M. T., Beal M. F. (2006). Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. PubMed DOI

Lowry O. H., Passonneau J. V. (1972).

Lynch M., Sayin U., Bownds J., Janumpalli S., Sutula T. (2000). Long-term consequences of early postnatal seizures on hippocampal learning and plasticity. PubMed DOI

Marella M., Seo B. B., Matsuno-Yagi A., Yagi T. (2007). Mechanism of cell death caused by complex I defects in a rat dopaminergic cell line. PubMed DOI

Nitecka L., Tremblay G., Charton G., Bouillot P., Berger M. L., Ben-Ari Y. (1984). Maturation of kainic acid seizure brain damage syndrome in the rat. II. Histopathological sequelae. PubMed

Ott M., Gogvadze V., Orrenius S., Zhivotovsky B. (2007). Mitochondria, oxidative stress and cell death. PubMed DOI

Parihar M. S., Parihar A., Villamena F. A., Vaccaro P. S., Ghafourifar P. (2008). Inactivation of mitochondrial respiratory chain complex I leads mitochondrial nitric oxide synthase to become pro-oxidative. PubMed DOI

Patel M. (2004). Mitochondrial dysfunction and oxidative stress: cause and consequence of epileptic seizures. PubMed DOI

Patel M., Li Q. Y. (2003). Age dependence of seizure-induced oxidative stress. PubMed DOI

Paxinos G., Watson C. (1998).

Perier C., Tieu K., Guégan C., Caspersen C., Jackson-Lewis V., Carelli V. (2005). Complex I deficiency primes Bax-dependent neuronal apoptosis through mitochondrial oxidative damage. PubMed DOI PMC

Rowley S., Patel M. (2013). Mitochondrial involvement and oxidative stress in temporal lobe epilepsy. PubMed DOI PMC

Ryan K., Backos D. S., Reigan P., Patel M. (2012). Post-translational oxidative modification and inactivation of mitochondrial complex I in epileptogenesis. PubMed DOI PMC

Ryan K., Liang L. P., Rivard C., Patel M. (2014). Temporal and spatial increase of reactive nitrogen species in the kainate model of temporal lobe epilepsy. PubMed DOI PMC

Sankar R., Shin D. H., Liu H., Mazarati A., Pereira de Vasconcelos A., Wasterlain C. G. (1998). Patterns of status epilepticus-induced neuronal injury during development and long-term consequences. PubMed PMC

Sipos I., Tretter L., Adam-Vizi V. (2003). Quantitative relationship between inhibition of respiratory complexes and formation of reactive oxygen species in isolated nerve terminals. PubMed DOI

Sullivan P. G., Dubé C., Dorenbos K., Steward O., Baram T. Z. (2003). Mitochondrial uncoupling protein-2 protects the immature brain from excitotoxic neuronal death. PubMed DOI PMC

Trotti D., Danbolt N. C., Volterra A. (1998). Glutamate transporters are oxidant-vulnerable: molecular link between oxidative and excitotoxic neurodegeneration? PubMed DOI

Waldbaum S., Patel M. (2010). Mitochondria, oxidative stress, and temporal lobe epilepsy. PubMed DOI PMC

Wasterlain C. G., Niquet J., Thompson K. W., Baldwin R., Liu H., Sankar R., et al. (2002). Seizure-induced neuronal death in the immature brain. PubMed DOI

Zielonka J., Kalyanaraman B. (2010). Hydroethidine- and MitoSOX-derived red fluorescence is not a reliable indicator of intracellular superoxide formation: another inconvenient truth. PubMed DOI PMC

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