Gelatinase B/lacZ Transgenic Mice, a Model for Mapping Gelatinase B Expression during Developmental and Injury-related Tissue Remodeling*
- From the Vision Research Laboratories, New England Medical Center, and the Departments of Ophthalmology and Anatomy, and Cell Biology, Tufts University School of Medicine, Boston, Massachusetts 02111
Abstract
Matrix metalloproteinases (MMPs) drive normal tissue remodeling and are implicated in a wide range of pathologies. Although MMP activity is controlled at multiple levels, the primary regulation of MMP activity is transcriptional. The transcriptional promoter elements required for MMP gene expression in cultured cells have been defined, but this has not been extended to the in vivo situation. In this paper, we show that the DNA sequences between −522 and +19 of the rabbit gelatinase B gene (MMP-9) (as characterized in the transgenic mouse line 3445) constitute a minimal promoter that drives appropriate developmental and injury-induced reporter gene expression in transgenic mice. We further show that the expression and activity of three transcription factors (NF-κB, AP-2, and Sp1) that control the activity of the gelatinase B promoter are selectively induced in the epithelium migrating to heal a wound. Although promoter activity parallels expression of the endogenous gene in cell cultures, we show by several criteria that cell cultures cannot model many aspects of promoter regulation in vivo. This study reveals that the transgenic mouse line 3445 might be a useful model for investigating the regulation of gelatinase B expressionin vivo and for identifying and characterizing new drugs that can control gelatinase B gene transcription.
Footnotes
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↵* This work was supported in part by Project Grant AR42981 from the National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵‡ Was on fellowship from the Italian government.
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↵§ Jules and Doris Stein Research to Prevent Blindness Professor. To whom correspondence should be addressed: Vision Research Laboratories, New England Medical Center, 750 Washington St., P. O. Box 450, Boston, MA 02111. Tel.: 617-636-9027; Fax: 617-636-4594; E-mail:efini{at}opal.tufts.edu.
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↵2 P. Bargagna-Mohan, K. J. Strissel, and M. E. Fini, submitted for publication.
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↵4 J. A. West-Mays, Z. Zhang, T. Nottoli, S. Hagopian-Donaldson, D. Libby, K. J. Strissel, and T. Williams, submitted for publication.
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↵3 R. Mohan and M. E. Fini, unpublished data.
- Abbreviations:
- ECM
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extracellular matrix
- MMP
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matrix metalloproteinase
- gelB
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gelatinase B
- TPA
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12-O-tetradecanoylphorbol-13-acetate
- PCR
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polymerase chain reaction
- β-gal
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β-galactosidase
- X-gal
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5-bromo-4-chloro-3-indolyl β-d-galactopyranoside
- PBS
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phosphate-buffered saline
- EMSA
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electrophoretic mobility shift assay.
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- Received May 11, 1998.
- Revision received July 9, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
