Changes In Expression Of GFRα1 In Animal Models Of Parkinson's Disease And The Cloning And Expression Of Its Gene

Objective:In recent years, nerve system diseases have trubled human health and impacted human life. Especially with the people's aging, the morbidity of Parkinson's disease, epilepsy, Alzheimer's disease, brain ischemia etc is growing. It has seriously effected on old people's quality of life. Many researchers focus on the treatment and mechanism of nerve system diseases. In recent years, it is found that GFRα1 was related with the onset process of Parkinson's disease, epilepsy, Alzheimer's disease, and cerebral ischemia. As the main receptor of the GDNF, GFRal has been a new focus of the treatment to nerve system diseases. In this experiment, the PD model was set up, and observed the change of the expression of GFRal in the striatum of rats.GFRα1 gene by cloning and GFRal protein from prokaryotic expression was gain,which provides experiment basis for the treatment of nerve system diseases and the research of the pathogenesisMethods:The Wistar rats were randomly selected to set up the experimental animal model. According to the double targets injection method, the right striatums of rats were injected with 6-OHDA.With stereotactic art, the PD animal model was set up. The successful PD model of rats were picked up for the HE dyeing, and immunohistochemistry analysis. Then the changes of quantity and shape of the neurons and the express quantity of GFRal were observed. To calculate the IOD with Image-pro6.0 to analysis the positive expression area, One-dimensional variance analysis was taken by SPSS 13.0.Total RNA of brain was gain from fetal rats. GFRα1-full-length sequence was amplified by RT-PCR. After the construction of the expression vector of pET-30a-GFRα1, it expressed in E.coli.Results:The PD rat model was successfully set up. By the immunohistochemical, the expression of GFRal in PD group of brain damaged side which were observed are decreased significantly, compared with the group of control,P<0.05.With RT-PCR, GFRal Full-length sequence were gain. After the expression in E.coli, a clear single protein band was identified by Western-blot.Conclusion:GFRal expression in PD model of the striatum of rats were decreased, compared with the control group, Successfully expressed GFRαl protein.