ABSTRACTTerminal erythroid differentiation is the process during which proerythroblasts differentiate to produceenucleated reticulocytes. While it is well established that during murine erythropoiesis in vivo, oneproerythroblast undergoes three mitosis to generate sequentially 2 basophilic, 4 polychromatic and 8orthochromatic erythroblasts, currently there is no method to quantitatively monitor this highly regulatedprocess. Here we outline a method that distinguishes each distinct stage of erythroid differentiation in cellsfrom mouse bone marrow and spleen based on expression levels of TER119, CD44 and cell size.Quantitative analysis revealed that the ratio of proerythroblasts:basophilic:polychromatic:orthromaticerythroblasts follows the expected 1:2:4:8 ratio, reflecting the physiological progression of terminalerythroid differentiation in normal mice. Moreover, in two stress erythropoiesis mouse models,phlebotomy-induced acute anemia and chronic hemolytic anemia due to 4.1R deficiency, the ratio of theseerythroblast populations remains the same as that of wild type bone marrow. In contrast, in anemicβ-thalassemia intermedia mice, there is altered progression which is restored to normal by transferrintreatment which has been previously shown to ameliorate the anemic phenotype. The means to quantitate invivo murine erythropoiesis using our approach is likely to have broad application in the study of alterederythropoiesis in various red cell disorders.

Quantitative analysis of murine terminal erythroid differentiation in vivo: novel method to study normal and disordered erythropoiesis

DE FRANCESCHI, Lucia;
2013

Abstract

ABSTRACTTerminal erythroid differentiation is the process during which proerythroblasts differentiate to produceenucleated reticulocytes. While it is well established that during murine erythropoiesis in vivo, oneproerythroblast undergoes three mitosis to generate sequentially 2 basophilic, 4 polychromatic and 8orthochromatic erythroblasts, currently there is no method to quantitatively monitor this highly regulatedprocess. Here we outline a method that distinguishes each distinct stage of erythroid differentiation in cellsfrom mouse bone marrow and spleen based on expression levels of TER119, CD44 and cell size.Quantitative analysis revealed that the ratio of proerythroblasts:basophilic:polychromatic:orthromaticerythroblasts follows the expected 1:2:4:8 ratio, reflecting the physiological progression of terminalerythroid differentiation in normal mice. Moreover, in two stress erythropoiesis mouse models,phlebotomy-induced acute anemia and chronic hemolytic anemia due to 4.1R deficiency, the ratio of theseerythroblast populations remains the same as that of wild type bone marrow. In contrast, in anemicβ-thalassemia intermedia mice, there is altered progression which is restored to normal by transferrintreatment which has been previously shown to ameliorate the anemic phenotype. The means to quantitate invivo murine erythropoiesis using our approach is likely to have broad application in the study of alterederythropoiesis in various red cell disorders.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11562/494755
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