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Blood, 1 October 2007, Vol. 110, No. 7, pp. 2718-2726.
Prepublished online as a Blood First Edition Paper on June 19, 2007; DOI 10.1182/blood-2006-04-016535.


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RED CELLS

Functional conservation of erythropoietin signaling in zebrafish

Noëlle Paffett-Lugassy1, Nelson Hsia1, Paula G. Fraenkel1, Barry Paw2, Irene Leshinsky3, Bruce Barut1, Nathan Bahary4, Jaime Caro3, Robert Handin2, and Leonard I. Zon1

1 Stem Cell Program and Division of Hematology/Oncology, Children's Hospital and Dana-Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute; 2 Department of Hematology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA; 3 Department of Medicine, Division of Hematology, Thomas Jefferson University, Jefferson Medical College, Philadelphia, PA; 4 Departments of Molecular Genetics & Biochemistry, and Medicine, Division of Oncology, University of Pittsburgh School of Medicine, PA

Erythropoietin (Epo) and its cognate receptor (EpoR) are required for maintaining adequate levels of circulating erythrocytes during embryogenesis and adulthood. Here, we report the functional characterization of the zebrafish epo and epor genes. The expression of epo and epor was evaluated by quantitative reverse transcriptase–polymerase chain reaction (RT-PCR) and whole-mount in situ hybridization, revealing marked parallels between zebrafish and mammalian gene expression patterns. Examination of the hypochromic mutant, weissherbst, and adult hypoxia-treated hearts indicate that zebrafish epo expression is induced by anemia and hypoxia. Overexpression of epo mRNA resulted in severe polycythemia, characterized by a striking increase in the number of cells expressing scl, c-myb, gata1, ikaros, epor, and ße1-globin, suggesting that both the erythroid progenitor and mature erythrocyte compartments respond to epo. Morpholino-mediated knockdown of the epor caused a slight decrease in primitive and complete block of definitive erythropoiesis. Abrogation of STAT5 blocked the erythropoietic expansion by epo mRNA, consistent with a requirement for STAT5 in epo signaling. Together, the characterization of zebrafish epo and epor demonstrates the conservation of an ancient program that ensures proper red blood cell numbers during normal homeostasis and under hypoxic conditions.


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