Toxic species of oxygen are generated by processes within lung cells or are formed in the extracellular milieu of the lung. Exposure of lung tissue to reactive oxygen species may induce cellular damage and, ultimately, respiratory distress. Enzymatic protection against such activated oxygen-based destruction is partially provided by catalase and superoxide dismutase (SOD). Mouse strain Csb possesses the only known nonhuman mammalian genetic variant of the catalase structural gene that produces a relatively diminished expression of catalase in the lung. This provides a unique probe to study the regulatory mechanisms active in normal lung catalase gene (Csa) expression. To understand the relationship between genetic variability of antioxidant gene expression and oxygen radical-induced respiratory distress, the molecular basis of the Csb-catalase mutation will first be determined, and lung regulatory mechanism(s) influenced by this genetic perturbation will be analyzed. In vitro translation and in vivo turnover studies of lung catalase mRNA and protein, will identify lung-specific control processes influenced by this mutation. Sequence analyses of catalase Csa and Csb cDNAs will identify any nucleotide differences between these lung mRNAs. To identify specific nucleotide sequences that produce Csb lung acatalasemia, recombinant expression plasmids containing hybrid catalase encoding molecules (Csa/Csb) will be tested for their ability to rescue acatalasemic fibroblasts from H2O2 toxicity. Hyperoxia will be used to induce oxidative stress to estimate the incidence, degree, and mechanisms of catalase and SOD induction in Csa and Csb lung tissue. Influence of lung maturational level and variability of gene expression of survival rates after oxidative stress will be evaluated. To identify regulatory sequences that may influence catalase gene expression in murine lung tissue, genomic catalase clones of 5'-noncoding sequences will be isolated and putative controlling regions tested for inductive capabilities. Results from this study will not only identify molecular regulatory processes which modulate lung antioxidant enzyme levels, but also provide an aspect of the molecular basis for susceptibility to the adult respiratory distress syndrome.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL039631-03
Application #
3356418
Study Section
(SRC)
Project Start
1987-09-30
Project End
1992-07-31
Budget Start
1989-08-01
Budget End
1990-07-31
Support Year
3
Fiscal Year
1989
Total Cost
Indirect Cost
Name
New York State Department of Health
Department
Type
DUNS #
002436061
City
Menands
State
NY
Country
United States
Zip Code
12204
Mossman, B T; Surinrut, P; Brinton, B T et al. (1996) Transfection of a manganese-containing superoxide dismutase gene into hamster tracheal epithelial cells ameliorates asbestos-mediated cytotoxicity. Free Radic Biol Med 21:125-31
Gibbs, L S; Del Vecchio, P J; Shaffer, J B (1992) Mn and Cu/Zn SOD expression in cells from LPS-sensitive and LPS-resistant mice. Free Radic Biol Med 12:107-11
Gallagher Jr, D S; Gibbs, L S; Shaffer, J B et al. (1992) Somatic cell mapping of bovine EC-SOD and SOD1L loci. Genomics 12:610-2
Tsan, M F; White, J E; Del Vecchio, P J et al. (1992) IL-6 enhances TNF-alpha- and IL-1-induced increase of Mn superoxide dismutase mRNA and O2 tolerance. Am J Physiol 263:L22-6
Del Vecchio, P J; Shaffer, J B (1991) Regulation of antioxidant enzyme expression in LPS-treated bovine retinal pigment epithelial and corneal endothelial cells. Curr Eye Res 10:919-25
Shaffer, J B; Treanor, C P; Del Vecchio, P J (1990) Expression of bovine and mouse endothelial cell antioxidant enzymes following TNF-alpha exposure. Free Radic Biol Med 8:497-502
Tsan, M F; White, J E; Treanor, C et al. (1990) Molecular basis for tumor necrosis factor-induced increase in pulmonary superoxide dismutase activities. Am J Physiol 259:L506-12
Shepard, B A; Shaffer, J B (1990) A rapid and convenient procedure for identifying recombinant plasmids. Biotechniques 8:388
Shaffer, J B; Preston, K E (1990) Molecular analysis of an acatalasemic mouse mutant. Biochem Biophys Res Commun 173:1043-50
Gibbs, L S; Shaffer, J B (1990) Nucleotide sequence of bovine copper/zinc superoxide dismutase cDNA generated by the polymerase chain reaction. Nucleic Acids Res 18:7171

Showing the most recent 10 out of 11 publications