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J Neurosci. 2005 Jun 15;25(24):5824-32. doi: 10.1523/JNEUROSCI.1192-05.2005.
2
GFP-tagged prion protein is correctly localized and functionally active in the brains of transgenic mice.绿色荧光蛋白标记的朊病毒蛋白在转基因小鼠大脑中定位正确且功能活跃。
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本文引用的文献

1
Subcellular localization of disease-associated prion protein in the human brain.疾病相关朊病毒蛋白在人脑中的亚细胞定位。
Am J Pathol. 2005 Jan;166(1):287-94. doi: 10.1016/S0002-9440(10)62252-3.
2
Prion genetics: new rules for a new kind of gene.朊病毒遗传学：一种新型基因的新规则。
Annu Rev Genet. 2004;38:681-707. doi: 10.1146/annurev.genet.38.072902.092200.
3
GFP-tagged prion protein is correctly localized and functionally active in the brains of transgenic mice.绿色荧光蛋白标记的朊病毒蛋白在转基因小鼠大脑中定位正确且功能活跃。
Neurobiol Dis. 2004 Aug;16(3):527-37. doi: 10.1016/j.nbd.2004.05.005.
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Identification and characterization of a novel mouse prion gene allele.一种新型小鼠朊病毒基因等位基因的鉴定与表征
Mamm Genome. 2004 May;15(5):383-9. doi: 10.1007/s00335-004-3041-5.
5
Mammalian prion biology: one century of evolving concepts.哺乳动物朊病毒生物学：一个概念不断演变的世纪。
Cell. 2004 Jan 23;116(2):313-27. doi: 10.1016/s0092-8674(03)01031-6.
6
Soluble dimeric prion protein binds PrP(Sc) in vivo and antagonizes prion disease.可溶性二聚体朊病毒蛋白在体内与PrP(Sc)结合并拮抗朊病毒病。
Cell. 2003 Apr 4;113(1):49-60. doi: 10.1016/s0092-8674(03)00201-0.
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Prion diseases: what is the neurotoxic molecule?朊病毒疾病：神经毒性分子是什么？
Neurobiol Dis. 2001 Oct;8(5):743-63. doi: 10.1006/nbdi.2001.0433.
8
Mutant prion proteins are partially retained in the endoplasmic reticulum.
J Biol Chem. 2001 Nov 9;276(45):42409-21. doi: 10.1074/jbc.M106928200. Epub 2001 Aug 29.
9
Tissue distribution of protease resistant prion protein in variant Creutzfeldt-Jakob disease using a highly sensitive immunoblotting assay.使用高灵敏度免疫印迹法检测变异型克雅氏病中蛋白酶抗性朊病毒蛋白的组织分布
Lancet. 2001 Jul 21;358(9277):171-80. doi: 10.1016/s0140-6736(01)05403-4.
10
Rapid cycling of lipid raft markers between the cell surface and Golgi complex.脂筏标志物在细胞表面和高尔基体复合体之间的快速循环。
J Cell Biol. 2001 Apr 30;153(3):529-41. doi: 10.1083/jcb.153.3.529.

在表达绿色荧光蛋白标记的朊病毒蛋白的转基因小鼠中朊病毒感染的可视化。

Visualization of prion infection in transgenic mice expressing green fluorescent protein-tagged prion protein.

作者信息

Barmada Sami J, Harris David A

机构信息

Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.

出版信息

J Neurosci. 2005 Jun 15;25(24):5824-32. doi: 10.1523/JNEUROSCI.1192-05.2005.

DOI:10.1523/JNEUROSCI.1192-05.2005
PMID:15958749
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6724869/
Abstract

Tg(PrP-EGFP) mice express an enhanced green fluorescent protein (EGFP)-tagged version of the prion protein (PrP) that behaves like endogenous PrP in terms of its posttranslational processing, anatomical localization, and functional activity. In this study, we describe experiments in which Tg(PrP-EGFP) mice were inoculated intracerebrally with scrapie prions. Although PrP-EGFP was incapable of sustaining prion infection in Tg(PrP-EGFP)/Prn-p(0/0) mice, it acted as a dominant-negative inhibitor that bound to, and fluorescently marked, deposits of PrPSc generated from endogenous PrP in Tg(PrP-EGFP)/Prn-p(+/+) mice. Scrapie infection of these latter animals caused a progressive accumulation of fluorescent PrP-EGFP aggregates in neuropil, axons, and prominently in the Golgi apparatus of neurons. Our results provide an entirely new picture of PrPSc localization during the course of prion infection, and they identify for the first time intracellular sites of PrPSc formation that are not well visualized with conventional immunohistochemical techniques.

摘要

转基因（PrP-EGFP）小鼠表达一种带有增强型绿色荧光蛋白（EGFP）标签的朊病毒蛋白（PrP），该蛋白在翻译后加工、解剖定位和功能活性方面与内源性PrP表现相似。在本研究中，我们描述了将羊瘙痒病朊病毒脑内接种到转基因（PrP-EGFP）小鼠体内的实验。虽然PrP-EGFP在转基因（PrP-EGFP）/Prn-p(0/0)小鼠中无法维持朊病毒感染，但它作为一种显性负性抑制剂，能与转基因（PrP-EGFP）/Prn-p(+/+)小鼠内源性PrP产生的PrPSc沉积物结合并进行荧光标记。对这些后一种动物进行羊瘙痒病感染，会导致荧光性PrP-EGFP聚集体在神经毡、轴突中逐渐积累，并且在神经元的高尔基体中显著积累。我们的结果提供了朊病毒感染过程中PrPSc定位的全新图景，并且首次确定了用传统免疫组织化学技术无法很好观察到的PrPSc形成的细胞内位点。