Abstract
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<.001%), a 1 min illumination of the cell to strong 470–490 nm light evoked a significant red fluorescence (excitation 520–550 nm, emission >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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
E. Takahashi, and K. Asano, Mitochondrial respiratory control compensates for slow oxygen diffusion in cardiomyocytes, Am. J. Physiol. 283, H871–H878 (2002).
E. Takahashi, H. Endoh, and K. Doi, Intracellular gradients of O2 supply to mitochondria in actively respiring single cardiomyocyte of rats, Am. J. Physiol. 276, H718–H724 (1999).
E. Takahashi, K. Sato, H. Endoh, Z. L. Xu, and K. Doi, Direct observation of radial intracellular Po2 gradients in a single cardiomyocyte of the rat, Am. J. Physiol. 275, H225–H233 (1998).
K. Groebe, An easy-to-use model for O2 supply to red muscle. Validity of assumptions, sensitivity to errors in data. Biophys. J. 68, 1246–1269 (1995).
E. Takahashi, and K. Doi, Visualization of oxygen level inside a single cardiac myocyte, Am. J. Physiol. 268, H2561–H2568 (1995).
M. B. Elowitz, M. G. Surette, P.-E. Wolf, J. Stock, and S. Leibler, Photoactivation turns green fluorescent protein red, Curr. Biol. 7, 809–812 (1997).
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Springer Science+Business Media, Inc.
About this paper
Cite this paper
Takahashi, E. et al. (2005). Genetic Oxygen Sensor. In: Okunieff, P., Williams, J., Chen, Y. (eds) Oxygen Transport to Tissue XXVI. Advances in Experimental Medicine and Biology, vol 566. Springer, Boston, MA. https://doi.org/10.1007/0-387-26206-7_6
Download citation
DOI: https://doi.org/10.1007/0-387-26206-7_6
Publisher Name: Springer, Boston, MA
Print ISBN: 978-0-387-25062-5
Online ISBN: 978-0-387-26206-2
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)