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Vol. 10, Issue 4, 935-945, April 1999
Department of Physiology, University of Massachusetts Medical
School, Worcester, Massachusetts 01605
We have developed a new approach to detect mechanical forces
exerted by locomoting fibroblasts on the substrate. Cells were cultured
on elastic, collagen-coated polyacrylamide sheets embedded with
0.2-µm fluorescent beads. Forces exerted by the cell cause deformation of the substrate and displacement of the beads. By recording the position of beads during cell locomotion and after cell
removal, we discovered that most forces were radially distributed, switching direction in the anterior region. Deformations near the
leading edge were strong, transient, and variable in magnitude, consistent with active local contractions, whereas those in the posterior region were weaker, more stable, and more uniform, consistent with passive resistance. Treatment of cells with cytochalasin D or
myosin II inhibitors caused relaxation of the forces, suggesting that
they are generated primarily via actin-myosin II interactions; treatment with nocodazole caused no immediate effect on forces. Immunofluorescence indicated that the frontal region of strong deformation contained many vinculin plaques but no apparent
concentration of actin or myosin II filaments. Strong mechanical forces
in the anterior region, generated by locally activated myosin II and transmitted through vinculin-rich structures, likely play a major role
in cell locomotion and in mechanical signaling with the surrounding environment.
Corresponding author. E-mail address:
yu-li.wang{at}ummed.edu.
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