The Behavioral And The Expressing Of CAMP Signaling Pathways Related Protein Expression In Hippocampus And Serum Of 5-HTT Knockout Mice

Objective:To observe the behavioral changes of 5-HTT（serotonin transporter）knockout mice,to study the differences of their psychological behavior,and to explore the possible role of c AMP signal pathway related protein level in the behavioral changes of 5-HTT knockout mice,so as to study the adverse psychological characteristics of accident prone drivers,such as anxiety,depression and tension,caused by 5-HTT gene polymorphism in the early stage of the research group To provide experimental basis for the pathogenesis.Methods:The experimental animals were divided into three groups:5-HTT^-/-（5-HTT expression disappeared）,5-HTT^+/-（5-HTT expression decreased）and 5-HTT^+/+（wild type）.10 mice in each group were compared and evaluated by open field experiment and forced swimming.Detection of c AMP protein in hippocampus,PKA protein in serum and AC protein in serum by ELISA,Western blot was used to detect the AC1,PKA,CREB and BDNF protein content in the hippocampus,and correlation analysis was used to explore the correlation between behavior and camp protein content in the hippocampus.Results:1.In the open field experiment,the scores of vertical activity,horizontal activity,the number of moving cells in the central area,the number of entering the central area and the activity time in the central area in the 5-HTT^-/-group were lower than those in the 5-HTT^+/-group and 5-HTT^+/+group（P<0.05）.However,the score of horizontal activity and the number of moving cells in the central area in the 5-HTT^+/-group were lower than those in the 5-HTT^+/+group（P<0.05）.2.The immobility time of 5-HTT^-/-group was longer than that of 5-HTT^+/-group and 5-HTT^+/+group in forced swimming,The immobility time of5-HTT^+/-group was also longer than that of 5-HTT^+/+group（P<0.05）.3.The results of ELISA showed that compared with the 5-HTT^+/+group,the hippocampal c AMP,serum PKA and serum AC contents of the 5-HTT^-/-group decreased（all P<0.05）,the hippocampal camp and serum PKA of the5-HTT^+/-group decreased（all P<0.05）;compared with the 5-HTT^+/-group,the hippocampal c AMP,serum PKA and serum AC contents of the 5-HTT^-/-group decreased（all P<0.05）.4.Western blot showed that compared with the 5-HTT^+/+group,the expression of AC1,PKA,CREB and BDNF in the hippocampus of the 5-HTT^-/-group decreased（all P<0.05）,and the expression of PKA in the 5-HTT^+/-group decreased（all P<0.05）;compared with the 5-HTT^+/-group,the expression of AC1 and BDNF in the hippocampus of the 5-HTT^-/-group decreased（P<0.05）.5.Correlation analysis results show that:（1）There was a positive correlation between the c AMP content in the hippocampus and the scores of vertical activity,horizontal activity,the number of moving cells in the central area,the length of staying in the center,the frequency of entering the center,the PKA content in the serum,and the AC content in the serum,with the correlation coefficients of 0.381,0.623,0.442,0.399,0.514,0.536,0.409,respectively,while the camp protein content in the hippocampus of the mice was negatively correlated with the immobility time of forced swimming The correlation coefficient was-0.663（all P<0.05）.（2）Serum PKA content was positively correlated with vertical activity score,horizontal activity score,number of moving cells in the central area,length of staying in the central area,frequency of entering the central area and serum AC content,with correlation coefficients of 0.634,0.662,0.509,0.566,0.606 and 0.362,respectively,while serum PKA content was negatively correlated with immobility time of forced swimming,with correlation coefficients of-0.459（all P<0.05）.（3）The serum AC content was positively correlated with the scores of vertical activity,horizontal activity,the number of moving cells in the central area,the length of staying in the center and the frequency of entering the center,with the correlation coefficients of 0.354,0.383,0.460,0.580 and 0.546,respectively,while the serum AC content was negatively correlated with the immobility time of forced swimming,with the correlation coefficients of-0.529（all P<0.05）.Conclusions:（1）5-HTT gene knockout mice showed lower curiosity and autonomy to the fresh environment,lower ability of inquiry autonomous activity,increased tension and anxiety,and more despair and depression under stress.（2）5-HTT gene knockout can reduce the expression level of c AMP signal transduction pathway related proteins in hippocampus and serum of mice,which makes the body prone to anxiety,depression and other adverse psychological behavior problems under stress.