Cell-matrix interactions in the avian corneal stroma: The use of anti-sense viral constructs to knock out β-containing integrins

The development and maintenance of corneal transparency requires the interaction of corneal fibroblasts and their precursors with specific extracellular matrix molecules through cellular receptors- βl-containing integrins are present on cells throughout corneal development and we hypothesize that the 3l integrin subunit is important in cell-matrix interactions during this time. To test this hypothesis, we have cloned the chicken &l subunit and constructed retroviral vectors containing sense and anti-sense βlcDNA along with a lacZ marker. High titers of recombinant virus containing both sense and anti-sense strands have been obtained and infection of chicken embryo fibroblasts is efficient based upon lacZ expression in in vitro assays. Preliminary studies have shown that cells infected with the anti-sense construct exhibit morphological variations in cell shape. Based on these results, we expect that βi expression will be reduced in corneal fibroblasts and periocular mesenchyme cells infected with the anti-sense construct. This retroviral system will provide a useful means of altering expression of integrin subunits in an effort to elucidate their functional interactions with matrix components during corneal development.