Genetic Oxygen Sensor
Genetic Oxygen Sensor
复制标题
基因氧传感器
DOI：
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发表时间：
2005
期刊：
影响因子：
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通讯作者：
Michihiko Sato
中科院分区：
文献类型：
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作者：
E. Takahashi;Tomohiro Takano;Aya Numata;Natsuho Hayashi;S. Okano;O. Nakajima;Y. Nomura;Michihiko Sato
本文报道了一种利用绿色荧光蛋白(GFP)进行体内氧测量的新方法。用聚乙烯亚胺试剂将表达载体pCMX-GFP瞬时导入COS7细胞，培养48h。在细胞暴露于缺氧气体(O2,580 nm)后，在光活化之前，缺氧气体是可以忽略的。这种(绿色)GFP荧光的红移在常氧条件下从未观察到。然后，我们在稳定表达GFP的转基因小鼠(绿色小鼠)中检验了这种方法的有效性。绿色荧光蛋白瞬时表达的小鼠心肌细胞的绿色荧光强度仅为∼的1/200时，所有分离的绿色小鼠心肌细胞均显示出明显的绿色荧光。缺氧条件下的光激活增强了这些细胞的红色荧光，但幅度远小于预期。综上所述，GFP可作为缺氧的原位探针。在表达GFP的转基因动物中，以亚微米空间分辨率在体内成像缺氧基因位点是可能的。
We report in this article a new method for in vivo oxygen measurement using green fluorescence protein (GFP). COS7 cells were transiently transfected with an expression vector, pCMX-GFP, using a polyethylenimine reagent and cultured for 48 hrs. After exposure of the cell to anoxic gas (O2 580 nm) that had been negligible before the photoactivation. This red shift of (green) GFP fluorescence was never observed in normoxia. We then examined the validity of this method in transgenic mice in which GFP is stably expressed (green mice). All the ventricular myocytes isolated from the green mice showed significant green fluorescence, although the intensity was ∼1/200 of the transiently GFP-expressing COS7 cells. The photoactivation in anoxia increased the red fluorescence in these cells, but the magnitude was much smaller than expected. In summary, GFP can be used as an in situ probe for hypoxia. In GFP-expressing transgenic animals, in vivo imaging of anoxic loci with a submicron spatial resolution may be possible.