期刊论文详细信息
Frontiers in Physiology
The Evolution of Erythrocytes Becoming Red in Respect to Fluorescence
Ulrich Boehm1  Petra Weissgerber1  Greta Simionato2  Lars Kaestner2  Stephan Quint2  Alexander Kihm2  Asena Abay2  Matthias W. Laschke3  Laura Hertz4  Sandra Ruppenthal4  Polina Petkova-Kirova4  Christian Wagner5 
[1] Center for Molecular Signaling (PZMS), Institute for Pharmacology, Saarland University, Homburg, Germany;Experimental Physics, Saarland University, Saarbrücken, Germany;Institute for Clinical and Experimental Surgery, Saarland University, Homburg, Germany;Institute for Molecular and Cell Biology, Saarland University, Homburg, Germany;Physics and Materials Science Research Unit, University of Luxembourg, Luxembourg City, Luxembourg;Theoretical Medicine and Biosciences, Saarland University, Homburg, Germany;
关键词: mouse model;    transfusion;    fluorescent protein;    intravital microscopy;    imaging;   
DOI  :  10.3389/fphys.2019.00753
来源: DOAJ
【 摘 要 】

Very young red blood cells, namely reticulocytes, can be quite easily recognized and labeled by cluster of differentiation antibodies (CD71, transferrin receptor) or by staining remnant RNA with thiazol orange. In contrast, age specific erythrocyte labeling is more difficult in later periods of their life time. While erythrocytes contain band 4.1 protein, a molecular clock, so far it has not been possible to read this clock on individual cells. One concept to track erythrocytes during their life time is to mark them when they are young, either directly in vivo or ex vivo followed by a transfusion. Several methods like biotinylation, use of isotopes or fluorescent labeling have proved to be useful experimental approaches but also have several inherent disadvantages. Genetic engineering of mice provides additional options to express fluorescent proteins in erythrocytes. To allow co-staining with popular green fluorescent dyes like Fluo-4 or other fluorescein-based dyes, we bred a mouse line expressing a tandem red fluorescent protein (tdRFP). Within this Brief Research Report, we provide the initial characterisation of this mouse line and show application examples ranging from transfusion experiments and intravital microscopy to multicolour flow cytometry and confocal imaging. We provide a versatile new tool for erythrocyte research and discuss a range of experimental opportunities to study membrane processes and other aspects of erythrocyte development and aging with help of these animals.

【 授权许可】

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