Antisense oligodeoxynucleotides targeting PDGF-B mRNA inhibit cell proliferation during embryonic rat lung development

Abstract

There is increasing evidence to suggest that platelet-derived growth factor (PDGF) or PDGF-like molecules play a role in fetal lung morphogenesis. Our previous studies demonstrated the presence of PDGF-AA and PDGF-BB homodimers in embryonic and fetal rat lung. To explore further the role for PDGF-BB in embryonic lung development, we conducted intervention studies using PDGF-B chain-specific antisense oligodeoxynucleotides in a simple embryonic rat lung explant system. Unmodified antisense PDGF-B oligodeoxynucleotides inhibited, in a concentration-dependent manner, DNA synthesis of embryonic lung. A maximal inhibition of 50% was observed. The inhibitory effect of antisense PDGF-B oligodeoxynucleotides on DNA synthesis was reversed by the addition of exogenous PDGF-BB but not PDGF-AA. Antisense treatment decreased PDGF-BB but not PDGF-AA protein content, as assessed by immunoblot analyses. Incubation of lung explants with PDGF-BB neutralizing antibodies also resulted in an inhibition of DNA synthesis. Morphometric analyses of antisense-treated cultures showed a significant reduction in lung size when compared to control cultures. The epithelial component of the embryonic lungs was specifically reduced, both in mass and DNA labelling index, by antisense treatment. The number of terminal buds of the lung explants was not significantly affected by antisense PDGF-B treatment. Scrambled PDGF-B oligodeoxynucleotides had no effect. These data suggest that PDGF-BB is involved in regulating growth, but not the degree of branching, of embryonic rat lung.