Blebbing dynamics during endothelial cell spreading.

Cell spreading is a critical component of numerous physiological phenomena including cancer metastasis, embryonic development, and mitosis. We have previously illustrated that cellular blebs appear after abrupt cell-substrate detachment and play a critical role in regulating membrane tension; however, the dynamics of bleb-substrate interactions during spreading remains unclear. Here we explore the role of blebs during endothelial cell spreading using chemical and osmotic modifications to either induce or inhibit bleb formation. We track cell-substrate dynamics as well as individual blebs using surface sensitive microscopic techniques. Blebbing cells (both control and chemically induced) exhibit increased lag times prior to fast growth. Interestingly, lamellae appear later for blebbing compared to non-blebbing cells, and in all cases, lamellae signal the start of fast spreading. Our results indicate that cellular blebs play a key role in the early stage of cell spreading, first by controling the initial cell adhesion and then by presenting a dynamic inhibition of cell spreading until a lamella appears and fast spreading ensues.