mouse model; transfusion; fluorescent protein; intravital microscopy imaging MONOMERIC RED; PROTEIN; C57BL/6; CALCIUM Physiology Physiology lars\_kaestner@me.com Wagner; Christian/A-1307-2009 Wagner; Christian/0000-0001-7788-4594 European Seventh Framework ProgramEuropean Union (EU) [602121] Deutsche Forschungsgemeinschaft (DFG)German Research Foundation (DFG) [Sonderforschungsbereich SFB894]; European Framework ``Horizon 2020'' [675115] This study was supported by the European Seventh Framework Program under grant agreement number 602121 (CoMMiTMenT; LK); the European Framework ``Horizon 2020'' under grant agreement number 675115 (RELEVANCE; CW and LK); and the Deutsche Forschungsgemeinschaft (DFG) Sonderforschungsbereich SFB894 (PW and UB). 32 0 1 1 Front. Physiol. IE0BO ISI:000472051700001
Abstract :
[en] 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.
Disciplines :
Physical, chemical, mathematical & earth Sciences: Multidisciplinary, general & others
Author, co-author :
Hertz, Laura; Kaestner, L (Reprint Author), Saarland Univ, Theoret Med Biosci, Homburg, Germany. Kaestner, L (Reprint Author), Saarland Univ, Expt Phys, Saarbrucken, Germany. Hertz, Laura