title: A novel approach for in vivo measurement of mouse red cell redox status.
abstract: Maintenance of a reducing redox balance is a critical physiologic function of red cells (RBC) that can be perturbed in variety of RBC pathologies. Here we describe a new approach to evaluate in vivo RBC redox status using a redox sensitive GFP (roGFP2) sensor under control of a β-globin mini-promoter, directing expression specifically to erythroid cells. RoGFP2 expressing RBCs demonstrate ratiometric and reversible shifts in fluorescence on exposure to oxidants and reductants. We demonstrate that roGFP2 expressing RBC can be used to monitor thiol redox status during in vitro phenylhydrazine treatment and over the course of in vivo RBC aging, where a shift to a more oxidized state is observed in older cells. Thus, roGFP2 transgenic mice are a new and versatile tool that can be used to probe how RBC redox status responds in the context of drug therapy, physiologic stressors and pathologic states.
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, LA Jolla, CA, USA.
authors.forename: Xiuling
authors.initials: X
authors.lastname: Xu
authors.forename: Katharina
authors.initials: K
authors.lastname: von Löhneysen
authors.forename: Katrin
authors.initials: K
authors.lastname: Soldau
authors.forename: Deborah
authors.initials: D
authors.lastname: Noack
authors.forename: Andrew
authors.initials: A
authors.lastname: Vu
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Friedman
mesh_terms: D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods; D058948:Cell Tracking / Q000379:methods*; D016922:Cellular Senescence / Q000502:physiology; D004909:Erythrocyte Indices / Q000502:physiology; D004912:Erythrocytes / Q000737:chemistry / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010084:Oxidation-Reduction
chemical_list: D049452:Green Fluorescent Proteins