Synaptic Mechanism Of Epilepsy

PART ONE: EXPRESSIONS OF SYNAPTIC VESICLE RELATED PROTEINS IN THE BRAIN TISSUE OF REFRACTORY EPILEPTIC PATIENTSObjective Based on the previous study of gene chips in refractory patients and kindling models, we detected the expression of genes and proteins related to synaptic vesicle recycling in the brains of refractory epileptic patients, to further explore the synapse function in human intractable epilepsy.Methods the temporal brain tissue from refractory epilepsy patients and head trauma patients were obtained. The refractory epilepsy patients were divided into two groups: the long illness duration and short illness duration groups, and the head trauma patients were used as control. RT-PCR, western blot, Immunohistochemistryand immunofluorescence were used to examine the mRNA expressions and protein expression of Dnm 1, Stx 1 and Syt 1.Results Dnm 1, Stx 1 and Syt 1 maily confined to the cytoplasm and cytomembrane. As compared with the control group, there appeared significant increased mRNA and protein level of Dnm 1 in long and short illness duration RE patients; there was no significant difference between 2 groups of RE. As to Stx 1 and Syt 1, the expressions were significantly up-regulated in long illness duration RE patients when compared with those in short illness duration RE and control., and there were no difference between hort illness duration RE and control.Conclusion The altered expressions of genes and proteins related to synaptic vesicle indicated that the synaptic visicle recycling was changed in the brains of RE. Dnm 1 is involved in the endocytosis of synaptic vesicle high levels of neuronal activity, our results suggested that Dyn 1 might be involved in the epileptogenesis of RE in the early stage through modulating endocytosis of neurotransmitter; Stx 1A is involved in vesicle fusion step and plays a key role in exocytosis and Syt 1 is involved in both exocytosis and endocytosis of synaptic vesicle. Our present results suggested that in the later stage of RE, both the exocytosis and endocytosis of synaptic vesicle were strengthened in refractory epileptic patients. Thus , the enhanced synaptic vesicle circulation may play an important role in the RE. PART TWO: THE CSF AND SERUM CONCENTRATION OF DNM 1, TALIN 2,Κ-IGLC,λ-IGLC IN POSTENCEPHALITIC EPILEPSY PATIENTSAim: A previous proteomics study found that talin 2 presented in the cerebrospinal fluid (CSF) of epileptic patients only but not in healthy controls . Talin 2 is found in focal cell adhesion and synapse and play role at those place.Methods: A total of 62 patients agreed to participate in the study. Dnm 1, Talin 2 were detected in the CSF and serum of and drug-effective post-encephalitic epilepsy(EPEE), drug-refractory post-encephalitic epilepsy( PEE), acute encephalitis with secondary seizures (AESS) acute encephalitis without secondary seizures (AE) patients using sandwich enzyme-linked immunosorbent assays.Results: A total of 62 patients agreed to participate in the study. The Dnm 1 concentrations of CSF and serum in AE, AESS,EPSS and RPEE had no significant difference(P>0.05);RPEE CSF talin 2 concentrations were greater when compared to AE, EPEE and AESS (p <0.05), While the serum talin 2 concentrations were significantly decreased in RPEE when compared with other 3 groups(p <0.05); the changes in CSFκ-IgLC,λ-IgLC and IgLCκ/λratio had no statistical difference among RPEE, EPEE, AESS and AE groups, but the changes in serumκ-IgLC,λ-IgLC and IgLCκ/λratio had extraordinary statistical difference: the serumκ-IgLC concentrations were significantly increased in RPEE; The serumλ-IgLC is significantly lower in RPEE when compared with other 3 groups and the concentrations in EPEE and AESS were significantly lower than that in AE; The serum IgLCκ/λratios in RPEE was significantly higher than other 3 groups and the ratios in EPEE and AESS were significantly higher than that in AE.Conclusions: We suggest that the differential concentration of CSF and serum talin 2 in RPEE is an intractability-related phenomenon but not a seizure-related phenomenon. The striking change of serumλ-IgLC andΚ/λratio in RPEE might be used as a diagnostic or prognostic parameter for RPEE and RPEE patients might be a target for immunoglobulin therapy.