Effects of acute seizures on cell proliferation, synaptic plasticity and long-term behavior in adult zebrafish.

Acute seizures may cause permanent brain damage depending on the severity. The pilocarpine animal model has been broadly used to study the acute effects of seizures on neurogenesis and plasticity processes and the resulting epileptogenesis. Likewise, zebrafish is a good model to study neurogenesis and plasticity processes even in adulthood. Thus, the aim of this study is to evaluate the effects of pilocarpine-induced acute seizures-like behavior on neuroplasticity and long-term behavior in adult zebrafish. To address this issue, adult zebrafish were injected with Pilocarpine (350 mg/Kg, i.p; PILO group) or Saline (control group). Experiments were performed at 1, 2, 3, 10 or 30 days after injection. We evaluated behavior using the Light/Dark preference, Open Tank and aggressiveness tests. Flow cytometry and BrdU were carried out to detect changes in cell death and proliferation, while Western blotting was used to verify different proliferative, synaptic and neural markers in the adult zebrafish telencephalon. We identified an increased aggressive behavior and increase in cell death in the PILO group, with increased levels of cleaved caspase 3 and PARP1 1 day after seizure-like behavior induction. In addition, there were decreased levels of PSD95 and SNAP25 and increased BrdU positive cells 3 days after seizure-like behavior induction. Although most synaptic and cell death markers levels seemed normal by 30 days after seizures-like behavior, persistent aggressive and anxiolytic-like behaviors were still detected as long-term effects. These findings might indicate that acute severe seizures induce short-term biochemical alterations that ultimately reflects in a long-term altered phenotype.

Effect of Memantine on Pentylenetetrazol-induced Seizures and EEG Profile in Animal Model of Cortical Malformation.

Developmental cortical malformations (DCM) are one of the main causes of refractory epilepsy. Many are the mechanisms underlying the hyperexcitability in DCM, including the important contribution of N-methyl-D-aspartate receptors (NMDAR). NMDAR blockers are shown to abolish seizures and epileptiform activity. Memantine, a NMDAR antagonist used to treat Alzheimers disease, has been recently investigated as a possible treatment for other neurological disorders. However, the effects on preventing or diminishing seizures are controversial. Here we aimed to evaluate the effects of memantine on pentylenetetrazole (PTZ)-induced seizures in the freeze-lesion (FL) model. Bilateral cortical microgyria were induced (FL) or not (Sham) in male Wistar neonate rats. At P30, subdural electrodes were implanted and 7 days later, video-EEG was recorded in animals receiving either memantine (FL-M or Sham-M) or saline (FL-S or Sham-S), followed by PTZ. Seizures were evaluated by video-EEG during one hour and scored according to Racine scale. The video-EEG analyses revealed that the number of seizures and the total duration of stage IV-V seizures developed during the 1 h-period increased after memantine application in all groups. The EEG power spectral density (PSD) analysis showed an increased PSD of pre-ictal delta in Sham-M animals and increased PSD of slow, middle and fast gamma oscillations after memantine injection that persists during the pre-ictal period in all groups. Our findings suggested that memantine was unable to control the PTZ-induced seizures and that the associated enhancement of PSD of gamma oscillations may contribute to the increased probability of seizure development in these animals.