Abstract
Ephrin-B3 is important in the regulation of cell proliferation, differentiation and migration via cell-cell contact, and can activate the reelin pathway during brain development. However, the effect of ephrin-B3 on hippocampal neurogenesis and the reelin pathway in epilepsy remains to be fully elucidated. In the present study, the expression of ephrin-B3 in pilocarpine-induced status epilepticus (SE) rats was investigated. SYBR Green-based reverse transcription-quantitative polymerase chain reaction analysis, immunohistochemical labeling and western blot analysis were used to detect the gene and protein expression levels of ephrin-B3 and reelin pathway proteins. Immunofluorescence staining of doublecortin (DCX) was utilized to analyze hippocampal neurogenesis. The data revealed that the mRNA and protein expression levels of ephrin-B3 in the hippocampus decreased during the spontaneous seizure period. Of note, the expression of reelin and its downstream phosphorylation disabled 1 (p-Dab1) were also notably decreased during the spontaneous seizure period, which showed similar dynamic changes as in the expression of ephrin-B3. In addition, it was found that the number of DCX-labeled neuronal progenitor cells was increased in the hippocampus following pilocarpine-induced SE. To further clarify the role of ephrin-B3 in neurogenesis and the reelin pathway in epilepsy, an exogenous ephrin-B3 clustering stimulator, EphB3-Fc, was infused into the bilateral hippocampus of the rats post-SE. Following EphB3-Fc injection, it was found that the expression levels of reelin and p-Dab1 were significantly increased in the epileptic rats following EphB3-Fc injection. The number of DCX-labeled neuronal progenitor cells was reduced in the hippocampus of the epileptic rats. Furthermore, the intensity and frequency of spontaneous recurrent seizures and electroencephalographic seizures were attenuated in the epileptic rats post-injection. These results demonstrated the critical role of ephrin-B3 in regulation of the reelin pathway and hippocampal neurogenesis in epilepsy, providing experimental evidence that ephrin-B3 functions as a potential protective factor in epilepsy, at least in animals.
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