Artículos de revistas
Significant Biochemical, Biophysical and Metabolic Diversity in Circulating Human Cord Blood Reticulocytes
Plos One. Public Library Science, v. 8, n. 10, 2013.
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Background: The transition from enucleated reticulocytes to mature normocytes is marked by substantial remodeling of the erythrocytic cytoplasm and membrane. Despite conspicuous changes, most studies describe the maturing reticulocyte as a homogenous erythropoietic cell type. While reticulocyte staging based on fluorescent RNA stains such as thiazole orange have been useful in a clinical setting; these 'sub-vital' stains may confound delicate studies on reticulocyte biology and may preclude their use in heamoparasite invasion studies. Design and Methods: Here we use highly purified populations of reticulocytes isolated from cord blood, sorted by flow cytometry into four sequential subpopulations based on transferrin receptor (CD71) expression: CD71high, CD71medium, CD71low and CD71negative. Each of these subgroups was phenotyped in terms of their, morphology, membrane antigens, biomechanical properties and metabolomic profile. Results: Superficially CD71high and CD71medium reticulocytes share a similar gross morphology (large and multilobular) when compared to the smaller, smooth and increasingly concave reticulocytes as seen in the in the CD71low and CD71negativesamples. However, between each of the four sample sets we observe significant decreases in shear modulus, cytoadhesive capacity, erythroid receptor expression (CD44, CD55, CD147, CD235R, and CD242) and metabolite concentrations. Interestingly increasing amounts of boric acid was found in the mature reticulocytes. Conclusions: Reticulocyte maturation is a dynamic and continuous process, confounding efforts to rigidly classify them. Certainly this study does not offer an alternative classification strategy; instead we used a nondestructive sampling method to examine key phenotypic changes of in reticulocytes. Our study emphasizes a need to focus greater attention on reticulocyte biology.810Singapore Ministry of Health NMRC [CBRG12nov036]SIgNHorizontal Programme on Infectious Diseases under the Agency for Science, Technology and Research (A*STAR, Singapore)Wellcome Trust of Great BritainOxford Tropical Medicine Research Programme of Wellcome Trust-Mahidol UniversityFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado do Amazonas (FAPEAM)National University of SingaporeNational University Health System, Singapore 'Start Up' GrantFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Singapore Ministry of Health NMRC [CBRG12nov036]FAPESP [2009/52013-3]