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The Mouse Resource for Pulmonary Disease has been evaluating the sensitivity of 11 different inbred strains of mice in models of allergic asthma (ovalbumin sensitization and challenge) and pulmonary fibrosis (intratracheal bleomyicn). To evaluate the potential relevance of genes inactivated by the Gene Trap Resource, we have prepared fixed lungs from unchallenged and challenged mice subjected to each protocol and, in collaboration with the In Situ Hybridization core, are evaluating the pattern of expression of these genes. For these initial experiments, we have included lungs from sensitive (A/J for ovalbumin and 129/SV for bleomycin) and resistant strains (C3H for ovalbumin and Balb/c for bleomycin) in both models.
We have also established the methods for evaluating global patterns of gene expression in the lungs of sensitive and resistant strains using oligonucleotide arrays composed of synthetic 70 mers. We are currently using arrays representing ~17,000 murine genes. Initial experiments comparing gene expression in lung and spleen have demonstrated the feasibility of this approach. We have evaluated patterns of gene expression in the lungs of saline challenged or ovalbumin challenged mice in B10D2, C57BL/6, AJ and C3H strains, and in the lungs of bleomycin treated mice in 129/SV and AJ strains. Array data of gene expression for lungs of animals from sensitive and resistant strains for each model can be viewed via microarray data of murine asthma and pulmonary fibrosis models.
In addition, we have begun to generate mice from selected Gene Trap ES cell lines to evaluate the roles of particularly promising candidate genes in lung development and in each of the models of lung disease. Thus far, we appear to have viable mice homozygous for insertions in the basement membrane protein, Nidogen 2, the tetraspanin TSPAN-3, the epithelial protein, Mfge8 and the cell adhesion molecule, JAM-1. To be certain about genotypes of these lines, putative homozygotes are being back-crossed to wild type mice. We are concurrently breeding animals in each line to evaluate in models of asthma and fibrosis and to perform detailed morphologic evaluation. We have obtained germline transmission for lines containing insertions in the TGFbeta signaling regulators, TAB1 and TAK1, and in the anion transporter, SLC38A2 (SAT1). These lines are currently being bred to obtain and evaluate mice homozygous for the genetrap insertions.