Phospho C-JUN is not involved in the neuronal apoptosis induced by glutamate on cultured cerebellar granule neurons

To investigate whether the phospho C JUN was involved in the neuronal apoptosis induced by glutamate.We established the model of apoptosis induced by glutamate on cultrured cerebellar granule neurons,identified apoptosis by using phase contrast microscopy,Hoechst33258 nucleus staining and DNA fragmentation agarose gel electrophoresis.After the neurons had been treated with glutamate(200 μmol/L),phospho C JUN was detected at different time point(5 min,30 min,1 h,2 h,4 h,8 h,16 h,and 24 h)by fluorescent immunocytochemistry in situ. We found that glutamate induced matured neuron's volume small,broke the neuronal dentrie and axon,and evoked the DNA fragmentation.The neuronal survival rate was 28 6%±5 2% after the neurons were exposed to glutamate for 24 h.At every time point,phospho C JUN positive neurons were not found in glutamate group,similar to depolarization medium(25 mmol/L KCl) group.Whereas,after the neurons were switched to re polarization medium(5 mmol/L KCl),many positive neurons were found.In addition,the density of fluorescence was the strongest at 4 h.After the neurons were treated for 4 h,the numbers of phospho C JUN positive neurons of re polarization group,depolarization group and glutamate group polarization were 124±17,8±3,and 5±3 respectively under the 400 times fluorescent microscopy.These results indicated that glutamate induced apoptosis on cultured cerebellar granule neurons and phospho C JUN might not be involved in the neuronal apoptosis. [