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Positron emission tomography imaging of conditional gene activation in the heart

lizongjin 添加于 2012-4-1 19:49 | 1113 次阅读 | 0 个评论

作者
Chang GY, Cao F, Krishnan M, Huang M, Li Z, Xie X, Sheikh AY, Hoyt G, Robbins RC, Hsiai T, Schneider MD, Wu JC
摘要
The Cre-loxP system has been routinely used for conditional activation and deletion of gene expression. However, the spatiotemporal manner of these events in the heart has not yet been defined by in vivo imaging. Adenovirus (1 x 10(9 )pfu) carrying the silent positron emission tomography (PET) reporter gene, herpes simplex virus type 1 thymidine kinase (HSV1-tk), was injected into the left ventricular wall of male transgenic mice (n=15) or FVB controls (n=8). Transgenic mice expressed Cre recombinase driven by a cardiac-specific alpha-myosin heavy chain (alpha-MHC) promoter. Following injection of the 9-[4-fluoro-3-(hydroxymethyl)butyl]guanine ([18F]-FHBG; 137+/-25 microCi) reporter probe, microPET imaging was used to assess the expression of HSV1-tk reporter gene in the myocardium. Two days following adenoviral injection, cardiac HSV1-tk gene activation resulted in tracer uptake of 3.20+/-0.51% ID/g for alpha-MHC-Cre and 0.05+/-0.02%ID/g for control mice (P<0.01). The in vivo results were confirmed by RT-PCR and Western blot analysis. Similar transfections were evaluated in both cardiac-specific and non-cardiac-specific cell lines. Enzyme activity showed a robust correlation (r2=0.82) between in vivo molecular imaging technique and traditional in vitro enzyme assays. With further development and validation, PET imaging will likely play an important role in the noninvasive, repetitive, and quantitative measurement of conditional gene activation in the future.
详细资料
- 关键词: Animals; Animals, Genetically Modified; *Gene Expression Regulation; Heart/*radiography; Integrases/metabolism; Male; Mice; *Positron-Emission Tomography; Promoter Regions, Genetic; Rats; Recombinant Proteins/metabolism; Transcriptional Activation; Ventricular Myosins/genetics/*metabolism
- 文献种类:期刊
- 期刊名称: Journal of Molecular and Cellular Cardiology
- 期刊缩写: J Mol Cell Cardiol
- 期卷页: 2007年第43卷第1期 18-26页
- 地址: The Molecular Imaging Program at Stanford (MIPS) and Department of Radiology, Stanford University, Palo Alto, CA, USA
- ISBN: 0022-2828
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