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
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