Research On The Relationship Between Lentivirus—mediated PTPRR Overexpression, Konckdown Mice And Depression

Objective:Through packaging high titer PTPRR gene over expression of virus particles and PTPRRsh RNA lentiviral particles in vitro, inject into mice hippocampus by mouse brain stereotactic surgery, build ptprr gene over expression mice and the ptprr gene knockdown mice model, compare depression related behavioral changes of tail suspension experiment, sucrose preference test, forced swimming in gene overexpression and gene knockdown mice, combine chronic not predict stimulation(CMS) to observe depression related behavior changes, investigate the relationship between the ptprr gene and depression in animal models.Methods:1. Transfecte successfully constructed ptprr gene overexpression vector, blank control GFP, PTPRR sh RNA carrier vector and the blank control sh GFP carrier vector into 293 T cells for packaging, condense to high titer virus particles in vitro.2. Microinject lentiviral particles in ince hippocampal by stereotaxic technique, resulting in ptprr gene overexpression and gene knockdown in mice hippocampus, take fresh hippocampus 28 days after the experiments, detect the PTPRR protein and PTPRR m RNA expression in mice hippocampus by Western blot, RT-PCR.3. 80 male C57 BL / 6J mice were divided into 10 groups, 8 rats in each group,(1) Lenti-sh PTPRR(ptprr gene knockdown) group;(2) lenti-sh GFP(low expression of empty vector group;(3) Lenti-PTPRR(the ptprr gene over expression) group.(4) lenti-GFP(empty vector overexpression) group;(5) control(normal control) group;(6) Lenti-sh PTPRR + CMS(ptprr gene knockdown intervention) group;(7) lenti-sh GFP+CMS(empty vector intervention) group;(8) the Lenti-PTPRR+CMS(ptprr gene overexpression intervention) group.(9) lenti-GFP+CMS(empty vector overexpression intervention) group;(10) control + CMS(normal control intervention) group. CMS intervention of the 6th – 10 th group 1 weeks after operation, observe the change of depression behavior after 21 days,Results:1. Successfully establish the specific PTPRR overexpression and PTPRRsh RNA virus particles, the virus titer reached 1x109 TU.2. 28 days after the injection of the virus found that Lenti-PTPRR mice hippocampus PTPRR m RNA and protein level were significantly increased compared with the control group and lenti GFP group(P<0.001); PTPRRm RNA and protein level of Lenti-PTPRRsh RNA mice hippocampus were significantly lower than that in the control group and Lenti-sh GFP group(P<0.001); and no significant difference were found between PTPRR m RNA and protein level in lenti GFP group and control group(P > 0.05).3. Significant differences were found in imbolity of each groups in forced swimming test(F=28.823), no significant difference were found between Lenti-GFP group and control group; no significant difference were found between Lenti-sh GFP group and control group(P>0.05); the immobility time in Lenti-PTPRR group was significantly longer than that in Lenti-GFP group(P<0.05); no significant change was found in the immobility time of forced swim test in Lenti-sh PTPRR group and Lenti-sh GFP(P>0.05); forced swim time in Lenti-PTPRR+CMS group were signicantly longer than that in Control+CMS group(P<0.001), forced swimming time in Lenti-sh PTPRR+CMS group was significantly reduced compared with Control+CMS group(P<0.001).4. In the tail suspension test, significant differences were found between the groups(F=10.215), no significant difference were found between Lenti-GFP group and control group, no significant difference were found between Lenti-sh GFP group and control group(P>0.05), no significant change were found between the immobility time in Lenti-PTPRR group and Lenti-GFP group. the immobility time in Lenti-PTPRR group were significantly decreased compared with the Lenti-GFP group. No significant change were found in immobility time of Lenti-sh PTPRR group and Lenti-sh GFP group(P>0.05), fouced swim time in Lenti-sh PTPRR+CMS group were signicantly decreased compared with the Control+CMS group(P<0.001).5. In the source intake test, significant differences were found between the groups(F=16.19), no significant difference were found between Lenti-GFP group and control group, no significant difference were found between Lenti-sh GFP group and control group(P>0.05), syrup consumption rate in Lenti-PTPRR group were significantly decresed compared with the Lenti-GFP group. No significant change were found in immobility time of Lenti-sh PTPRR group and Lenti-sh GFP group(P>0.05), syrup consumption rate in Lenti-sh PTPRR+CMS group were signicantly decreased compared with the Control+CMS group(P<0.001).Conclusions:1. The PTPRR overexpression and PTPRRsh RNA lentiviral particles vector and packaging particles are successfully established.;2. The mice model of PTPRR gene overexpression and gene knockdown was successfully established.;3. The overexpression of PTPRR can induce the increasement of depressive behavior in mice, and the overexpression of PTPRR gene may be more sensitive to stress..4. PTPRR gene knockdown may improve the despair behavior of depressed mice.