帕金森病α-Synuclein转基因小鼠模型的建立  

Establishment of α-Synuclein Transgenic Mouse Model of Parkinson's Disease

作　　者：高宁[1] 全雄志[2] 陈炜[2] 李万波[2] 刘鹏[1] 冯娟[1] 张连峰[2] 秦川[1] 

机构地区：[1]中国医学科学院实验动物研究所,北京协和医学院比较医学中心,卫生部人类疾病比较医学重点实验室,北京100021 [2]中国医学科学院实验动物研究所,北京协和医学院比较医学中心遗传中心,北京100021

出　　处：《中国比较医学杂志》2008年第5期28-31,36,I0005,共6页Chinese Journal of Comparative Medicine

基　　金：北京市转基因平台建设项目TC2006-02(Z0006303041231);中央级公益性科研院所基本科研务费专项基金DWS200704

摘　　要：目的建立人α-Synuclein野生型及两个突变型A53T和A30P基因的转基因动物模型,研究α-Synuclein在帕金森病发病过程中的作用。方法构建人α-Synuclein表达载体,利用显微注射法制备人α-Synuclein基因的转基因小鼠。通过PCR方法鉴定转基因首建鼠及其子代基因型。通过RT-PCR和Western blotting方法鉴定转基因小鼠脑组织中人α-Synuclein mRNA和蛋白表达情况。采用免疫组化鉴定人α-Synuclein在小鼠脑组织中的表达情况。通过Rotatingrod实验评价转基因小鼠的行为改变情况。结果得到表达水平不同的野生型人α-Synuclein转基因小鼠2个品系。得到表达水平不同的A53T突变型α-Synuclein转基因小鼠2个品系。得到表达水平不同的A30P突变型α-Synuclein转基因小鼠3个品系。免疫组化显示,转基因小鼠大脑海马、新皮层、纹状体区出现人α-Synuclein阳性标记的细胞。Rotating rod实验结果显示转基因小鼠表现出明显的进行性运动能力障碍。结论建立了转人α-Synuclein基因的帕金森病小鼠模型。Objective To establish human α-Synuclein wild type gene, A53T mutation and A30P mutation transgenic animal models and investigate the role of α-Synuclein in the pathogenesis of Parkinson＇s disease. Methods The PDGF-h-α-Synuclein vectors expressing the α-Synuclein wild type gene, A53T mutation and A30P mutation were constructed. The transgenic mice were created with the microinjection method. The genotype of transgenic founders and its filial generations were identified by PCR. The expression level of human α-Synuclein was analyzed by RT-PCR and Western blotting. The expression level of human α- Synuclein in the brain of transgenic mice was analyzed by immunohistochemistry staining. The decline in motor performance of transgenic mice was measured by rotating rod experiment. Results Two human α-Synuclein WT transgenic lines with different expression levels were established. Two human α-Synuclein A53T transgenic lines with different expression levels were established. The three human α-Synuelein A30P transgenie lines with different expression levels were established either. The expression of α- Synuelein was detected in hippocampus, neocortex and substantia nigra of all the 7 lines by immunohistochemistry staining. All of the transgenie mice lines showed obviously progressive decline in motor performance by rotating rod experiment. Conclusion The human α-Synuelein transgenie mouse models of Parkinson＇s disease were established.

关 键 词：转基因模型 Α-SYNUCLEIN 帕金森病 突变 

分 类 号：Q813[生物学—生物工程]