After status epilepticus (SE), actin cytoskeleton (F-actin) becomes progressively deconstructed in the hippocampus, which is consistent with the delayed pyramidal cell death in both time course and spatial distribution. A variety of experiments show that calcineurin inhibitors such as FK506 are able to inhibit the SE-induced actin depolymerization. However, it is still unclear what changes happen to the F-actin in the epileptic brain after FK506 treatment. A pilocarpine model of SE in mice was used to examine the effects of FK506 on the F-actin in the hippocampal neurons. The post SE (PSE) mice with or without FK506 treatment were monitored consecutively for 14 days to examine the frequency and duration of spontaneous seizures. The effects of FK506 on the activity of cofilin and actin dynamics were assessed at 7 and 14 d PSE by western blots. The organization of F-actin, neuronal cell death, and glial reactions were investigated by phalloidin staining, histological and immunocytochemical staining, respectively. As compared to the PSE + vehicle mice, FK506 treatment significantly decreased the frequency and duration of spontaneous seizures. Relative to the PSE + vehicle mice, western blots detected a partial restoration of phosphorylated cofilin and a significant increase of F/G ratio in the hippocampus after FK506 treatment. In the PSE + vehicle mice, almost no F-actin puncta were left in the CA1 and CA3 subfields at 7 and 14 d PSE. FK506-treated PSE mice showed a similar decrease of F-actin, but the extent of damage was significantly ameliorated. Consistently, the surviving neurons became significantly increased in number after FK506 treatment, relative to the PSE + vehicle groups. After FK506 treatment, microglial reaction was partially inhibited, but the expression of GFAP was not significantly changed, compared to the PSE + vehicle mice. The results suggest that post-epileptic treatment with FK506 ameliorated, but could not stop the deconstruction of F-actin or the delayed neuronal loss in the PSE mice.
Keywords: FK506; Filamentous actin; Pilocarpine; Status epilepticus.
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