Synphilin-1转基因小鼠模型的制备及帕金森病相关蛋白的表达

摘要
图/表
参考文献
相关文章 (0)

全文: PDF (2937 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要目的：探讨Synphilin-1与帕金森病发病机制的相关性。方法：利用朊蛋白（Prp）启动子载体将人类Synphilin-1基因结合注入到小鼠受精卵中制作转基因（Tg）小鼠。通过小鼠尾活组织DNA的PCR检测确定小鼠基因型。然后进行PD相关蛋白免疫印迹分析，脑组织切片免疫组织化学分析，双标记免疫荧光染色及Tg小鼠表型与运动能力分析。结果：Synphilin-1广泛表达于包括黑质的Tg小鼠大脑的神经元中，但没有观察到多巴胺（DA）神经元的大量流失，Synphilin-1蛋白在Tg小鼠的大脑中被多泛素化，部分呈现不溶性。利用改良SHIRPA法观察Tg组小鼠与非Tg组小鼠在行为和形态学上的改变，差异无统计学意义。通过加速转圈测试和足迹分析观察2组小鼠的运动能力，Tg组小鼠与非Tg组小鼠相比运动能力减少。结论：Synphilin-1可能参与运动功能的调节，但不影响脑组织内PD相关蛋白的表达，对DA神经元亦无显著影响。

	服务
	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	金洪国
	唐维国
	金美玉
	何松彬
	周敏
	寇雪莲

关键词 ： Synphilin-1；帕金森病；转基因小鼠；抗酪氨酸羟化酶；&alpha, -共核蛋白

Abstract：Objective: To explore the pathogenesis correlafion of Synphilin-1 with Parkinson’s disease. Methods: Generated transgenic mice expressing human Synphilin-1 under the prion protein prom-oter. The genotype of offspring mice was determined by a PCR assay of DNA obtained from tail biopsies.The human Synphilin-1 gene was injected into the fertilized ovum using prion promoter vector to generate the transgenic mice, then PD-related protein immunoblot analysis, the brain tissue slice analysis with immunohistochemistry, double labeling immunofluorescence staining and the phenotypic and athletic analysis of transgenic mice were performed. Results: Synphilin-1 was widely expressed in neurons in the brain, including the substantianigra, but massive loss of dopamine neurons was not observed. In the transgenic mouse brain, Synphilin-1 protein was polyubiquitinated and partially insoluble. Although modified-SHIRPA revealed no significant difference in behavior and morphology, the reduced rotarod performance and step length were observed in transgenic mice as compared with non-transgenic littermates. Conclusion: Synphilin-1 may be involved in motor function, and its accumulation in the central nervous system can cause motor impairments. Synphilin-1 may be involved in regulation of motor function, but had no significant effect of the expression of Parkinson’s disease related proteins nor dopamine neurons in the brain tissue.

Key words： Synphilin-1 Parkinson’s disease transgenic mice tyrosine hydroxylase;α-Synuclein

收稿日期: 2015-01-14

基金资助:浙江省科技厅钱江人才计划项目（2010R10078）。

通讯作者: 唐维国，主任医师，Email：tangweiguo2003@163.com。

作者简介: 金洪国（1974-），男，吉林延边人，副主任医师，博士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2015/V45/I9/660

[1] Engelender S, Kaminsky Z, Gu X, et al. Synphilin-1 associates with α-synuclein and promotes the formation of cytosolic inclusions[J]. Nat Genet, 1999, 22(1): 110-114.
[2] Wakabayashi K, Engelender S, Yoshimoto M, et al. Synphilin-1 is present in Lewy bodies in Parkinson’s disease[J]. Ann Neurol, 2000, 47(4): 521-523.
[3] Szargel R, Rott R, Engelender S. Synphilin-1 isoforms in Parkinson’s disease: regulation by phosphorylation and ubiquitylation[J]. Cell Mol Life Sci, 2008, 65(1): 80-88.
[4] Szargel R, Rott R, Eyal A, et al. Synphilin-1A inhibits seven in absentia homolog (SIAH) and modulates alpha-synuclein monoubiquitylation and inclusion formation[J]. J Biol Chem, 2009, 284(17): 11706-11716.
[5] Sutachan JJ, Casas Z, Albarracin SL, et al. Cellular and molecular mechanisms of antioxidants in Parkinson’s disease[J]. Nutr Neurosci, 2012, 15(3): 120-126.
[6] Tanji K, Mori F, Kito K, et al. Synphilin-1-binding protein NUB1 is colocalized with nonfibrillar, proteinase K-resistant α-synuclein in presynapses in Lewy body disease[J]. J Neuropathol Exp Neurol, 2011,70(10): 879-889.
[7] Marx FP, Soehn AS, Berg D, et al. The proteasomal subunit S6 ATPase is a novel synphilin-1 interacting protein-implications for Parkinson’s disease[J]. FASEB J, 2007, 21(8): 1759-1767.
[8] Hoshino T, Kawase Y, Okamoto M, et al. Cutting edge: IL-18-transgenic mice: in vivo evidence of a broad role for IL-18 in modulating immune function[J]. J Immunol, 2001, 166(12): 7014-7018.
[9] Fukuba H, Takahashi T, Jin HG, et al. Abundance of aspargynyl-hydroxylase FIH is regulated by Siah-1 under normoxic conditions[J]. Neurosci Lett, 2008, 433(3): 209-214.
[10] Masuya H, Inoue M, Wada Y, et al. Implementation of the modified-SHIRPA protocol for screening of dominant phenotypes in a large-scale ENU mutagenesis program[J]. Mamm Genome, 2005, 16(11): 829-837.
[11] Goldberg MS, Fleming SM, Palacino JJ, et al. Parkin-deficient mice exhibit nigrostriatal deficits but not loss of dopaminergic neurons[J]. J Biol Chem, 2003, 278(44): 43628-43635.
[12] Masliah E, Rockenstein E, Veinbergs I, et al. Dopaminergic loss and inclusion body formation in α-synuclein mice: implications for neurodegenerative disorders[J]. Science,
2000, 287(5456): 1265-1269.
[13] Antony PM, Diederich NJ, Balling R. Parkinson’s disease mouse models in translational research[J]. Mamm Genome,2011, 22(7-8): 401-419.
[14] Yamashita H, Matsumoto M. Molecular pathogenesis, experimental models, and new therapeutic strategies for Parkinson’s disease[J]. Regen Med, 2007, 2(4): 447-455.
[15] 周辉, 盛汉松, 张弩, 等. 体外诱导分化的多能成体祖细胞移植到帕金森病大鼠模型后的形态学观察[J]. 温州医学院学报, 2011, 41(5): 431-435.
[16] Koppula S, Kumar H, More SV, et al. Recent updates in redox regulation and free radical scavenging effects by herbal products in experimentalmodels of Parkinson’s disease[J].Molecules, 2012, 17(10): 11391-11420.