Open AccessA Double in vivo Biotinylation Technique for Objective Assessment of Aging and Clearance of Mouse Erythrocytes in Blood Circulation
Author(s) -
Rajiv K. Saxena,
Nitin Bhardwaj,
Sumedha Sachar,
Niti Puri,
Sanjay Khandelwal
Publication year - 2012
Publication title -
transfusion medicine and hemotherapy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.971
H-Index - 39
eISSN - 1660-3818
pISSN - 1660-3796
DOI - 10.1159/000342524
Subject(s) - biotinylation , phosphatidylserine , flow cytometry , in vivo , red blood cell , autofluorescence , biology , biotin , microbiology and biotechnology , biochemistry , genetics , fluorescence , phospholipid , physics , quantum mechanics , membrane
We have recently developed a new technique to objectively identify erythrocyte cohorts of defined age in mouse blood. The technique (termed double in vivo biotinylation, DIB) involves an initial biotinylation of all erythrocytes in circulation, followed after a few days by a second biotinylation, at a lower density, that labels the biotin-negative erythrocytes that have entered since the first biotinylation. The proportions of biotin(high), biotin(low), and biotin(negative) erythrocytes are enumerated by flow cytometry. The DIB technique allows us to track age-related changes on erythrocyte cohorts (Protocol A), and to simultaneously identify very young and older erythrocyte populations in the blood (Protocol B). Using this technique, we have reexamined: i) the relationship between age and buoyant density of erythrocytes, ii) erythrocyte destruction through a random removal mechanism, and iii) the expression of phosphatidylserine on aging erythrocytes. We have also used the DIB technique to study age-related changes in the expression of various markers like CD47 and CD147 and green autofluorescence in aging erythrocyte populations.