Podosome formation in the murine palatal mucosae: Its proteolytic role in rete peg formation

Background: Basement membrane remodeling is an indispensable factor for oral mucosal rete peg formation, but how the basement membrane is remodeled remains unclear. Our previous study indicated that keratinocyte growth factor induces the assembly of podosomes, which are dynamic organelles critical for matrix remodeling in human immortalized oral epithelial cells. This study explores podosome formation and its role in basement membrane remodeling during murine oral mucosal rete peg formation.

Methods: Perinatal murine palatal tissue slices were obtained from embryonic day 17.5 (E 17.5) to postnatal day 10.5 (P 10.5) BALB/c mice. Rete peg formation was observed by hematoxylin and eosin (HE) staining. Proteolysis of the basement membrane was detected by immunofluorescence staining. The assembly of podosomes and their correlation with basement membrane proteolysis were investigated by laser scanning confocal microscopy.

Results: The shape of basal layer keratinocytes at the sites of emerging rete pegs changed from typically polygonal to spindle-shaped. Basement membrane proteolysis, indicated by decreased type IV collagen (Col IV) staining, was detected during rete peg formation. Classical markers for podosomes, including cortactin/Tks5, WASP, and matrix metalloproteinase foci, were easily observed at the spindle-shaped cells. Podosomes were visible in regions where there was a significant decrease in Col IV staining.

Conclusions: These observations indicated that podosomes form at the front of the emerging rete peg and may play a pivotal role in basement membrane remodeling during rete peg formation.