Expression And Action Of GalR2 Gene In The Mice Model Of Depression

In this study, a eukaryotic expression plasmid pEGFP -GalR2 was constructed, the expression of GalR2 gene in the level of transcription and translation was observed and the intracellular location of GalR2 gene productions was identified. We established the C57BL/6J mice model of depression, and evaluated the influence of expression of GalR2 gene.Partâ… The expression and identification of GalR2 geneObjective: To obtain full length coding sequence of GalR2 gene and construct a eukaryotic expression vector containing the fusion gene of mouse GalR2 and EGFP, and observe its expression and location in Hela cells with fluorescence microscope.Methods: The coding sequence of the GalR2 gene was acquired by RT-PCR from the total RNA of C57BL/6J mouse hippocampus, in which EcoRâ… ,BamHâ… restrict endonuclease site in up and down primer was added respectively. The GalR2 gene was cloned into pEGFP-C1 to construct recombinant plasmid pEGFP-GalR2, identified by PCR, restrict endonuclease digestion and DNA sequencing. Then the pEGFP-GalR2 was transiently transfected into Hela cells, and the expression of GalR2 gene was detected by RT-PCR, Western blotting and fluorescence microscope. Result: The gene about 1116bp was successfully obtained as expected. The eukaryotic plasmid pEGFP-GalR2 successfully constructed. The GalR2 gene expression product in Hela cells was detected by RT-PCR and Western blotting. The fusion protein GalR2-EGFP was localized in cell nucleus with fluorescence microscope.Conclusion: The eukaryotic expression plasmid pEGFP-GalR2 was successfully constructed. The fusion protein can be coexpressed in Hela cells and its localization was in cell nucleus.Partâ…¡The establishment of mice model of depression and discussion of the influence of GalR2 expression against this modelObjective: To establish the C57BL/6J mice model of depression and evaluate the influence of GalR2 gene expression on depression.Methods: After preliminary screening using weighting, openfield test and resistance-to-capture test,C57BL/6J mice(n=35)were randomly divided into four groups:â… .CUMS (chronic unpredictable mild stress) group (n=10);â…¡.CUMS+CORT (cortisol) group (n=10);â…¢.CORT (characterize the effect of repeated corticosterone) group (n=10);â…£.Normal control group (n=5). We took notes on body weighting and food intaking every day. After 28 days, the mice were then tested for changes in fearful/anxious/depression-like behavior in liquid consumption test and forced swim test. The C57BL/6J mice of depression (n=7) that fromâ… andâ…¢groups were randomly divided into 2 groups( pEGFP-GalR2 group and pEGFP-C1 group). After surgery 48h, all animals were tested behavior in the forced swim test compared with pre-operation.Result: The mice model of depression has been established successfully at 28th day. Body weight, food intake, sugar consumption and the percentage of preference for sugar were significantly decreased and pure water consumption was increased compared with control group. In forced swim test, the depressed mice spent less time on swimming and struggling, and more time on immobiliting.â…¡group can not tolerate the combination of the two models, in the short term, body weight of mice rapidly loss and death. Compare withâ… group, the mice fromâ…¢had more changes in liquid consumption test and forced swim test.After surgery, the mice number of the pEGFP-GalR2 group was survived 3 and died 1; and the pEGFP-C1 group was survived 2 and died 1. The statistically significant were not found by forced swim test compared with pEGFP-C1 group and pre-operation.Conclusion: The mice model of depression has been established successfully. The combination of CUMS and CORT was too seriously to bear, the mice died after a short period of time. The model of CORT was better than CUMS. The actual effect of GalR2 protein on depression must be verified by experiment in future.