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Prepublished online as a Blood First Edition Paper on September 12, 2002; DOI 10.1182/blood-2002-02-0569.
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Blood, 1 February 2003, Vol. 101, No. 3, pp. 1155-1163
PHAGOCYTES
A macrophage colony-stimulating factor receptor-green
fluorescent protein transgene is expressed throughout the mononuclear
phagocyte system of the mouse
R. Tedjo Sasmono,
Delvac Oceandy,
Jeffrey W. Pollard,
Wei Tong,
Paul Pavli,
Brandon J. Wainwright,
Michael C. Ostrowski,
S. Roy Himes, and
David A. Hume
From the Institute for Molecular Bioscience and ARC
Special Research Centre for Functional and Applied Genomics, University
of Queensland, Brisbane, Australia; the Albert Einstein
College of Medicine of Yeshiva University, Bronx, NY; The Canberra
Hospital, Woden, Australian Capital Territory, Australia;
and the Department of Molecular Genetics, The Ohio State University,
Columbus, OH.
The c-fms gene encodes the receptor for macrophage
colony-stimulating factor (CSF-1). The gene is expressed selectively in the macrophage and trophoblast cell lineages. Previous studies have
indicated that sequences in intron 2 control transcript elongation in
tissue-specific and regulated expression of c-fms. In
humans, an alternative promoter was implicated in expression of the
gene in trophoblasts. We show that in mice, c-fms
transcripts in trophoblasts initiate from multiple points within the
2-kilobase (kb) region flanking the first coding exon. A reporter gene
construct containing 3.5 kb of 5' flanking sequence and the downstream
intron 2 directed expression of enhanced green fluorescent protein
(EGFP) to both trophoblasts and macrophages. EGFP was detected in
trophoblasts from the earliest stage of implantation examined at
embryonic day 7.5. During embryonic development, EGFP highlighted the
large numbers of c-fms-positive macrophages, including
those that originate from the yolk sac. In adult mice, EGFP
location was consistent with known F4/80-positive macrophage
populations, including Langerhans cells of the skin, and permitted
convenient sorting of isolated tissue macrophages from disaggregated
tissue. Expression of EGFP in transgenic mice was dependent on intron 2 as no lines with detectable EGFP expression were obtained where either
all of intron 2 or a conserved enhancer element FIRE (the
Fms intronic regulatory element) was removed. We
have therefore defined the elements required to generate myeloid- and
trophoblast-specific transgenes as well as a model system for the study
of mononuclear phagocyte development and function.
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