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Blood, 1 September 2004, Vol. 104, No. 5, pp. 1291-1297.
Prepublished online as a Blood First Edition Paper on March 2, 2004; DOI 10.1182/blood-2003-09-3105.
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HEMATOPOIESIS
The pu.1 promoter drives myeloid gene expression in zebrafish
Karl Hsu,
David Traver,
Jeffery L. Kutok,
Andreas Hagen,
Ting-Xi Liu,
Barry H. Paw,
Jennifer Rhodes,
Jason N. Berman,
Leonard I. Zon,
John P. Kanki, and
A. Thomas Look
From the Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA; Division of Hematology/Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA; Division of Hematology/Oncology, Children's Hospital Boston, Harvard Medical School, Boston, MA; Department of Pathology, Brigham and Women's Hospital, Boston, MA; and Division of Hematology, Brigham & Women's Hospital, Boston, MA.
PU.1 is a member of the Ets family of transcription factors and plays an essential role in the development of both myeloid and lymphoid cells. To examine zebrafish pu.1 (zpu.1) expression in subpopulations of blood cells during zebrafish development, we linked a 9-kb zebrafish genomic fragment upstream of the zpu.1 initiator codon to green fluorescent protein (GFP) and microinjected this construct to generate stable transgenic lines. GFP-positive fluorescent myeloid precursors were observed migrating from the anterolateral mesoderm in living embryos from 16 to 28 hours after fertilization (hpf) in a pattern that overlaps the expression pattern of endogenous zpu.1 mRNA. Analysis of larval histologic sections revealed GFP-expressing hematopoietic cells in the developing zebrafish kidney. Flow cytometric analysis of cells from adult whole kidney marrow revealed 2 discrete subpopulations of GFP-positive cells, which after cell sorting exhibited either myeloid or early lymphoid morphology. Thus, the zebrafish zpu.1 promoter fragment used here is capable of driving reporter gene expression in subsets of embryonic and adult hematopoietic cells. These transgenic lines will be useful to dissect the cellular and molecular control of myeloid cell differentiation, and this promoter fragment may prove useful in the development of zebrafish models of acute myeloid leukemia.
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