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Journal of Histochemistry and Cytochemistry, Vol. 51, 941-949, July 2003, Copyright © 2003, The Histochemical Society, Inc.

ARTICLE

Analysis of the Mouse CSF-1 Gene Promoter in a Transgenic Mouse Model

Sherry L. Abbouda, Maria Bunegina, Nandini Ghosh–Choudhurya, and Kathleen Woodruffa
a South Texas Veteran's Health Care System, Audie L. Murphy Division, and the Department of Pathology, University of Texas Health Science Center, San Antonio, Texas

Correspondence to: Sherry L. Abboud, Dept. of Pathology, U. of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78284. E-mail: abbouds@uthscsa.edu

CSF-1 stimulates monocyte and osteoclast populations. However, the molecular mechanisms involved in regulating CSF-1 gene expression are unclear. To identify regulatory regions that control normal CSF-1 gene expression, a -774/+183-bp fragment of the murine CSF-1 promoter was analyzed in vitro and in vivo. Transcriptional activity was high in cultured osteoblasts that express CSF-1 mRNA compared to ARH-77 B cells that lack CSF-1 gene expression. Transient transfection of osteoblasts with promoter deletion constructs showed that the -774-bp fragment conferred the highest transcriptional activity and contained activator and repressor sequences. To assess the ability of the CSF-1 promoter to confer normal tissue expression of CSF-1, transgenic mice containing the -774/+183-bp region driving the E. coli ß-galactosidase (lacZ) reporter gene were generated. ß-Gal analysis of whole tissue extracts showed transgene expression in all tissues tested except liver and kidney. At the cellular level, the pattern of ß-gal expression in the spleen, thymus, bone, lung, and testes of adult transgenic mice mimicked normal endogenous CSF-1 mRNA expression in non-transgenic littermates detected by in situ hybridization. This region also directed appropriate transgene expression to sites in other tissues known to synthesize CSF-1, with the exception of the liver and kidney. These findings indicate that the -774-bp fragment contains cis-acting elements sufficient to direct CSF-1 gene expression in many tissues. CSF-1 promoter/lacZ mice may be useful for studying the transcriptional mechanisms involved in regulating CSF-1 gene expression in tissues throughout development.

(J Histochem Cytochem 51:941–949, 2003)

Key Words: macrophage colony- stimulating factor, gene expression, transgenic mice


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