Targeting gene expression to the vascular wall in transgenic mice using the murine preproendothelin-1 promoter.
Open Access
- 1 March 1995
- journal article
- Published by American Society for Clinical Investigation in JCI Insight
- Vol. 95 (3), 1335-1344
- https://doi.org/10.1172/jci117784
Abstract
To develop a system for overexpressing genes in the vascular wall, we created transgenic mice using the reporter gene luciferase and the murine preproendothelin-1 promoter. In vitro analysis suggested that the murine 5'-flanking region contained endothelial-specific elements in a 5.9-kb fragment. Five transgenic mice colonies established from independent founders all exhibited the highest level of luciferase activity in the aorta with up to 8,540 light units per microgram of protein. Immunohistochemistry with anti-luciferase antisera revealed high levels of expression in the endothelial cells of both large and small arteries and lower levels of expression in veins and capillaries. Significant expression was also seen in arterial smooth muscle cells and in select epithelial surfaces which is consistent with the known distribution of endothelin-1 in mammals. The further demonstrate the targeting capability of this system, we overexpressed the lipid-peroxidating enzyme, human 15-lipoxygenase, in the vessel wall of transgenic mice. As with luciferase, expression of active enzyme and immunohistochemical localization in vascular cells were documented in transgenic animals. Hence, this new system can be used to direct expression of molecules to the vascular wall for the purpose of examining the biological significance of either overexpression or inhibition of select proteins.Keywords
This publication has 33 references indexed in Scilit:
- Glomerular epithelial cells synthesize endothelin peptides.1992
- cDNA cloning and chromosomal assignment of the gene encoding endothelin 3Journal of Biological Chemistry, 1989
- A NOVEL PEPTIDE, VASOACTIVE INTESTINAL CONTRACTOR, OF A NEW (ENDOTHELIN) PEPTIDE FAMILY - MOLECULAR-CLONING, EXPRESSION, AND BIOLOGICAL-ACTIVITY1989
- Endothelin: visualization of mRNAs by in situ hybridization provides evidence for local action.Proceedings of the National Academy of Sciences, 1989
- Structural Organization and Chromosomal Assignment of the Gene Encoding EndothelinJournal of Biological Chemistry, 1989
- The human endothelin family: three structurally and pharmacologically distinct isopeptides predicted by three separate genes.Proceedings of the National Academy of Sciences, 1989
- Formation of 15-hydroxyeicosatetraenoic acid (15-HETE) as the predominant eicosanoid in aortas from Watanabe Heritable Hyperlipidemic and cholesterol-fed rabbitsAtherosclerosis, 1988
- Molecular cloning and primary structure of human 15-lipoxygenaseBiochemical and Biophysical Research Communications, 1988
- A novel potent vasoconstrictor peptide produced by vascular endothelial cellsNature, 1988
- A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye BindingAnalytical Biochemistry, 1976