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Blood, 15 December 2001, Vol. 98, No. 13, pp. 3618-3625
HEMATOPOIESIS
Human granulocyte-macrophage colony-stimulating factor
(hGM-CSF) stimulates primitive and definitive
erythropoiesis in mouse embryos expressing hGM-CSF
receptors but not erythropoietin receptors
Hiroaki Hisakawa,
Daisuke Sugiyama,
Ichiko Nishijima,
Ming-jiang Xu,
Hong Wu,
Kazuki Nakao,
Sumiko Watanabe,
Motoya Katsuki,
Shigetaka Asano,
Ken-ichi Arai,
Tatsutoshi Nakahata, and
Kohichiro Tsuji
From the Departments of Clinical Oncology, Molecular
and Developmental Biology, and Molecular Medicine, Division of DNA
Biology and Embryo Engineering, Center for Experimental Medicine, The
Institute of Medical Science, The University of Tokyo, Japan;
Department of Pediatrics, Kochi Medical School, Nankoku, Japan; the
Department of Pediatrics, Graduate School of Medicine, Kyoto
University, Japan; and the Howard Hughes Medical Institute and
Department of Molecular and Medical Pharmacology, University of
California at Los Angeles School of Medicine.
Although erythropoietin (EPO) and its receptor (EPOR) are crucial
for the proliferation, survival, and terminal differentiation of
erythroid progenitors, it remains to be elucidated whether EPOR-unique
signaling is required for erythropoiesis. To address this issue, human
granulocyte-macrophage colony-stimulating factor (hGM-CSF) receptor
(hGMR)-transgenic mice and heterozygous EPOR mutant mice were crossed
by in vitro fertilization. In methylcellulose clonal culture of fetal
liver (FL) cells of generated hGMR-expressing EPOR / embryos at embryonic day (E) 12.5 of
gestation, hGM-CSF stimulated erythroid colony formation under
serum-containing and serum-free conditions. Analysis of globin
expression in individual erythrocyte-containing colonies formed from
E12.5 FL cells showed that hGM-CSF supports primitive and definitive
erythropoiesis even in EPOR/ embryos. In comparison of
activities between hGM-CSF and EPO in hGMR-expressing
EPOR+/+ embryos, the 2 substances supported the formation
of similar numbers of erythroid colonies in clonal culture of E12.5 FL
cells; enhanced adult, but not embryonic, globin synthesis; and induced increase of GATA-1 expression and decrease of erythroid Kruppel-like factor and cMyb expression in the FL cells. On the other hand, in E8.0
yolk sac erythropoiesis, both substances had a similar effect on
erythroid colony formation, but hGM-CSF induced an increase of
 -major globin expression, while EPO did not. All together, the
results of the present study demonstrated that hGM-CSF can stimulate
the proliferation and differentiation of primitive and definitive
erythroid cells independently of EPOR signal if they express hGMR, and
the activity is comparable to that of EPO in definitive, but not
primitive, erythropoiesis.
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