| Objective Through the establishment of primary rat hippocampal neuron system in vitro,the relationship between Sema3 F and CREB at gene and transcriptional level was studied.Method 1.The bilateral hippocampi of SD rats within 24 hours after birth were isolated under the aseptic condition of the super-clean table,and the primary rat hippocampal neurons were cultured.2.Experimental group: primary rat hippocampal neurons were extracted and Sema3 F,was added to the culture medium to take samples for 0 and 30 minutes respectively.Control group: primary rat hippocampal neurons were extracted,the same concentration of fetal bovine serum was added to the culture medium,and samples were taken for 0 and 30 minutes,respectively.3.The changes of gene and mRNA expression in primary hippocampal neurons of experimental group and control group were detected by high-throughput sequencing technology,and compared to the known reference genome by diamond software,mapping gene was obtained,and then differential genes were screened.According to the previous research results of the research group and consulting a large number of related literatures about neuronal axon guidance,synaptic reconstruction,growth cone collapse and so on,we further selected CREB as the target gene from the differential genes for further research.The FPKM value of differential genes was calculated.SPSS22.0 statistical software was used for statistical analysis,the data obtained by t test,P < 0.05.The difference was statistically significant.Results The expression of CREB DNA and CREB mRNA in the primary hippocampal neurons of the experimental group has increased after the addition of Sema3 F,while there was no significant change in the expression of CREB DNA and CREB mRNA in the primary hippocampal neurons of the control group.Conclusion 1.Sema3 F can promote the expression of CREB DNA in primary rat hippocampal neurons;2.Sema3 F can promote the expression of CREB mRNA in primary rat hippocampal neurons at the transcriptional level. |
