Screening differentially expressed genes involved in apoptosis of primary cultured rat cerebellar granule neurons by mRNA differential display RT-PCR

AIM: To Screen and identify differentially expressed genes that involved in apoptosis model in rat cerebellar granule neurons (CGNs).METHODS: The rat cerebellar granule neurons were isolated and primarily cultured.Fluorescent differential display RT-PCR (FDD RT-PCR) was performed to screen differentially expressed ESTs in the apoptosis model of primarily cultured rat CGNs.ESTs were subcloned into pGEM-T EasyTM vector and then sequenced.Alignment assay in non-redunant database was applied for encoding information.Reverse Northern blotting was used to appraise the results from DDRT-PCR.RESULTS: 164 pieces of differentially expressed ESTs were obtained by FDDRT-PCR.17 of them were subcloned and sequenced.5 ESTs of 17 were confirmed to be positive results by reverse Northern blotting.CONCLUSION: DD-PCR is a rapid,simple-operation and sensitive method for screening differentially expressed genes,which would contribute to the molecular mechanisms of apoptosis/survive of CGNs.