Contact dynamics during keratocyte motility.
BACKGROUND: Keratocytes are specialised, rapidly moving cells that generate substantial contractile force perpendicular to their direction of locomotion. Potential roles for contractile force in cell motility include cell-body transport, regulation of adhesion, and retraction of the cell's trailing edge. RESULTS: To investigate contact dynamics, we used simultaneous confocal fluorescence and interference reflection microscopy to image keratocytes injected with fluorescent vinculin. We found that contacts formed behind the leading edge and grew beneath both the lamellipodium and the cell body. Contacts in the middle of the cell remained stationary relative to the substrate and began to disassemble as the cell body passed over them. In contrast, contacts in the lobes of the cell grew continuously and more rapidly, incorporated more vinculin, and slid inwards towards the sides of the cell body. Contact sliding often led to merging of contacts before their removal from the substrate. CONCLUSIONS: We suggest a synthesis of two existing, apparently conflicting models for keratocyte motility, in which network contraction progressively reorients actin filaments using the contacts as pivots, forming bundles that then generate lateral tension by a sliding-filament mechanism. Contact dynamics vary between the middle of the cell and the lobes. We propose that laterally opposed contractile forces first enhance contact growth and stability, but escalating force eventually pulls contacts from the substrate at the back of the cell, without interfering with the cell's forward progress.