Erythroblasts from friend virus infected- and phenylhydrazine-treated mice accurately model erythroid differentiation.
The dynamics of gene expression during terminal erythroid differentiation have been examined in three murine models; the erythroleukaemia cell line HCD-57 and splenic erythroblasts isolated from mice treated with either the anaemia-inducing strain of Friend virus (FVA cells) or the haemolytic agent phenylhydrazine (PHZ cells). In response to erythropoietin (EPO) and haemin, HCD-57 cells proliferated and synthesized haemoglobin, but failed to complete terminal differentiation as indicated by lack of change in both gene expression and morphological appearance. In contrast, EPO-induced terminal differentiation in FVA and PHZ cells in vitro was accompanied by increases in haemoglobin positivity, morphological maturation and a shared pattern of gene expression. EPO receptor (EPO-R) mRNA levels peaked before globin gene expression which was maximal at 24 h. Peak GATA-1 and EKLF mRNA levels also preceded the globin gene peak, but the highest NF-E2 levels coincided with maximal globin levels, suggesting a role for NF-E2 in the maintenance, rather than the initiation of globin gene expression. Peak expression of delta-aminolaevulinic acid synthase (ALAS) coincided with peak globin expression. FVA and PHZ cells represent more effective models than the HCD-57 cell line for the investigation of erythroid gene expression during EPO-regulated terminal erythropoiesis.
Hodges
V M
VM
Department of Haematology, The Queen's University of Belfast, Royal Victoria Hospital, Belfast, Northern Ireland. vhodges@ic.ac.uk
Winter
P C
PC
Lappin
T R
TR
D000818:Animals; D048429:Cell Size; D004900:Erythroblasts / Q000187:drug effects / Q000473:pathology* / Q000821:virology; D004920:Erythropoiesis / Q000187:drug effects; D004921:Erythropoietin / Q000494:pharmacology; D005622:Friend murine leukemia virus / Q000502:physiology*; D015870:Gene Expression; D006454:Hemoglobins / Q000378:metabolism; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology* / Q000821:virology*; D051379:Mice; D010659:Phenylhydrazines / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D006454:Hemoglobins; D010659:Phenylhydrazines; C030299:phenylhydrazine; D004921:Erythropoietin