动物造模,帕金森病大鼠模型 z-bio 2022-11-03 294

　　Purpose: Founder α⁃ Parkinson's disease model of transgenic rats with synaptonuclein A30P mutant

　　Methods: Using lentivirus system to construct overexpression wild type α⁃ SYN carrier pLKO ⁃ CMV ⁃ α⁃ SYN ⁃ WT ⁃ P2A ⁃ GFP and α⁃ SYNA30P mutation vector pLKO ⁃ CMV ⁃ α⁃ SYN ⁃ A30P ⁃ P2A ⁃ GFP was transfected into 293FT cells respectively, and WB was detected 24h after transient transfection α⁃ SYN expression level. After being packaged and concentrated by lentivirus, the virus diluent was injected into the dense part of substantia nigra of the rat's midbrain using stereotaxic technology to overexpress α⁃ SYN wild type and A30P mutant lentivirus particles immunofluorescence staining α⁃ The distribution of SYN and tyrosine hydroxylase (TH), and the changes in the number of dopaminergic neurons in the substantia nigra compacta of the midbrain were observed. Rotatingrod test was used to evaluate the behavior changes of rats injected with A30P lentivirus

　　Results: The wild type and mutant A30P gene expression vectors were highly expressed in 293FT cells α⁃ SYN protein TH immunofluorescence results showed that: compared with virus diluent group α⁃ SYN wild type and A30P mutant rats can reduce the number of dopaminergic neurons in substantia nigra compacta of midbrain, while α⁃ SYNA30P lentivirus injection group had more neurons in the substantia nigra of the midbrain, with a significant difference. Immunofluorescence of the brain neuron deletion site of A30P transgenic rats found that TH staining in this area was almost negative, α⁃ SYN protein shows a large amount of aggregation, which indicates that the disappearance of brain neurons is accompanied by α⁃ The aggregation of SYN protein and the sharp decrease of TH expression further reveal overexpression α⁃ SYN A30P can lead to the reduction and degeneration of the number of dopaminergic neurons. In addition, the results of rotatingod experiment show that it is overexpressed α⁃ SYNA30P rats showed obvious progressive motor dysfunction

　　Conclusion: Human is established through lentivirus system α⁃ The PD model of SYNA30P mutant transgenic rats lays a foundation for the research of PD pathogenesis and drug development