Objective Munc18 is considered as an intracellular protein that plays an important role in exocytosis of *** studies have demonstrated the presence of autoantibodies against Munc18 in a subgroup of Rasmussen’s enceph...
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Objective Munc18 is considered as an intracellular protein that plays an important role in exocytosis of *** studies have demonstrated the presence of autoantibodies against Munc18 in a subgroup of Rasmussen’s encephalitis ***,the machinery of Munc18 autoimmunity is still *** present study was aimed to investigate Munc18 release from primary cultured neurons,Munc18 distribution on the outer plasma membrane of neurons,and the neurotoxicity of Munc18 *** The cerebral cortical neurons from embryonic day 17 SpragueDawley rats were prepared and cultured in neurobasal *** proteins in culture medium were precipitated with 10% trichloroacetic acid,and analyzed by *** proteins on neuronal surface were biotinylated with EZ-Link-sulfoNHS-LC-Biotin,and collected with avidin-conjugated agarose beads followed by immunoblotting *** cell surface immunofluorescent staining,the living neurons were labeled with anti-Munc18 antibody at 4 °*** injury was assessed by lactate dehydrogenase（LDH） *** Munc18 was detected in culture medium by immunoblotting *** treatment with 50 μmol/L glutamate for 1 h,Munc18 content in medium was ***,β-actin and syntaxin1 were not detected in culture medium,and LDH release was not significantly ***,glutamate enhanced Munc18 distribution on outer plasma *** neuron staining also demonstrated the localization of Munc18 on neuronal surface after glutamate treatment,especially at contacting regions between ***-induced increase of surface Munc18 distribution was suppressed by NMDA receptor antagonist MK801,but not by AMPA receptor antagonist ***,compared with c-Fos antibody,Munc18 antibody could induce neuronal injury,when culture medium contained the components of *** A portion of Munc18 can be released from neurons or distributed on neuronal surface,which can be enha
Objective To determine if DNA excision repair enzymes oxoguanine glycosylase 1 （OGG1） and xeroderma pigmentosum group F protein （XPF） are involved in the pathogenesis of Parkinson＇s disease （PD） in a cell model...
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Objective To determine if DNA excision repair enzymes oxoguanine glycosylase 1 （OGG1） and xeroderma pigmentosum group F protein （XPF） are involved in the pathogenesis of Parkinson＇s disease （PD） in a cell model. Methods PC12 cells were treated with 1-Methyl-4-phenylpyridine ion （MPP＋） for various periods of time to induce oxidative DNA damage. MTT assay was used to determine cell viability. Immunocytochemistry with antibody against 8-hydroxy-2＇- deoxyguanosine （8-oxodG） was used to evaluate oxidative DNA damage. Immunoblotting was used to detect the protein levels of OGG1 and XPF. Results MPP＋ treatment （1 mmol/L） for 18 h and 24 h reduced cell viability to 78.6% and 70.3% of the control, respectively, in a time-dependent way. MPP＋ increased the immunoreactivity of 8-oxodG in the cytoplasm at 3 h and in the nucleus at 24 h of treatment. With the treatment of MPP＋, the expression of OGG1 was significantly increased at 1 h, reaching a peak at 3 h, and then it was decreased at 24 h, as compared to that with vehicle treatment. The same effect was exerted on XPF level, except that the XPF level reached a peak at 18 h of MPP＋ treatment. Moreover, the maximally-increased protein level of OGG1 by MPP＋ was approximately 2-fold higher than that of XPF. Conclusion MPP＋ treatment could time- dependently induce increases in OGG1 and XPF expressions in PC12 cells. Also, this study indicates that the base and nucleotide excision repair pathways may be compensatorily activated in the early stage of pathogenesis in the cells after MPP＋ treatment.
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