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Published 8 July 2002. doi:10.1083/jcb.200203022
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© The Rockefeller University Press, 0021-9525/2002/7/31 $5.00
The Journal of Cell Biology, Volume 158, Number 1, July 8, 2002 31-37

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Dual-wavelength fluorescent speckle microscopy reveals coupling of microtubule and actin movements in migrating cells



Wendy C. Salmon, Michael C. Adams and Clare M. Waterman-Storer

Department of Cell Biology and Institute for Childhood and Neglected Diseases, The Scripps Research Institute, La Jolla, CA 92037

Address correspondence to Clare M. Waterman-Storer, Department of Cell Biology and Institute for Childhood and Neglected Diseases, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037. Tel.: (858) 784-9764. Fax: (858) 784-9779. E-mail: waterman{at}scripps.edu

Interactions between microtubules (MTs) and filamentous actin (f-actin) are involved in directed cell locomotion, but are poorly understood. To test the hypothesis that MTs and f-actin associate with one another and affect each other's organization and dynamics, we performed time-lapse dual-wavelength spinning-disk confocal fluorescent speckle microscopy (FSM) of MTs and f-actin in migrating newt lung epithelial cells. F-actin exhibited four zones of dynamic behavior: rapid retrograde flow in the lamellipodium, slow retrograde flow in the lamellum, anterograde flow in the cell body, and no movement in the convergence zone between the lamellum and cell body. Speckle analysis showed that MTs moved at the same trajectory and velocity as f-actin in the cell body and lamellum, but not in the lamellipodium or convergence zone. MTs grew along f-actin bundles, and quiescent MT ends moved in association with f-actin bundles. These results show that the movement and organization of f-actin has a profound effect on the dynamic organization of MTs in migrating cells, and suggest that MTs and f-actin bind to one another in vivo.

Key Words: live cell microscopy; cytoskeleton; cell motility; dynamic instability; retrograde flow


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