title: Quantitative analysis of murine terminal erythroid differentiation in vivo: novel method to study normal and disordered erythropoiesis.
abstract: Terminal erythroid differentiation is the process during which proerythroblasts differentiate to produce enucleated reticulocytes. Although it is well established that during murine erythropoiesis in vivo, 1 proerythroblast undergoes 3 mitosis to generate sequentially 2 basophilic, 4 polychromatic, and 8 orthochromatic erythroblasts, currently there is no method to quantitatively monitor this highly regulated process. Here we outline a method that distinguishes each distinct stage of erythroid differentiation in cells from 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:orthromatic erythroblasts follows the expected 1:2:4:8 ratio, reflecting the physiologic progression of terminal erythroid differentiation in normal mice. Moreover, in 2 stress erythropoiesis mouse models, phlebotomy-induced acute anemia and chronic hemolytic anemia because of 4.1R deficiency, the ratio of these erythroblast populations remains the same as that of wild-type bone marrow. In contrast, in anemic β-thalassemia intermedia mice, there is altered progression which is restored to normal by transferrin treatment which was previously shown to ameliorate the anemic phenotype. The means to quantitate in vivo murine erythropoiesis using our approach will probably have broad application in the study of altered erythropoiesis in various red cell disorders.
authors.affiliation: Red Cell Physiology Laboratory, New York Blood Center, New York, NY, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.forename: Jianhua
authors.initials: J
authors.lastname: Zhang
authors.forename: Yelena
authors.initials: Y
authors.lastname: Ginzburg
authors.forename: Huihui
authors.initials: H
authors.lastname: Li
authors.forename: Fumin
authors.initials: F
authors.lastname: Xue
authors.forename: Lucia
authors.initials: L
authors.lastname: De Franceschi
authors.forename: Joel Anne
authors.initials: JA
authors.lastname: Chasis
authors.forename: Narla
authors.initials: N
authors.lastname: Mohandas
authors.forename: Xiuli
authors.initials: X
authors.lastname: An
mesh_terms: D000740:Anemia / Q000097:blood / Q000188:drug therapy / Q000473:pathology; D000818:Animals; D001854:Bone Marrow Cells / Q000473:pathology / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation / Q000379:methods / Q000592:standards; D018450:Disease Progression; D004900:Erythroblasts / Q000473:pathology* / Q000502:physiology*; D004920:Erythropoiesis / Q000502:physiology; D005260:Female; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D006412:Hematopoietic Stem Cells / Q000473:pathology / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D015203:Reproducibility of Results; D013154:Spleen / Q000473:pathology / Q000502:physiology; D014168:Transferrin / Q000494:pharmacology; D017086:beta-Thalassemia / Q000097:blood* / Q000235:genetics / Q000473:pathology*
chemical_list: D014168:Transferrin