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Vol. 11, Issue 9, 2999-3012, September 2000
Department of Pathology, Centre Médical Universitaire,
Geneva, Switzerland
Migrating cells are polarized with a protrusive lamella at the cell
front followed by the main cell body and a retractable tail at the rear
of the cell. The lamella terminates in ruffling lamellipodia that face
the direction of migration. Although the role of actin in the formation
of lamellipodia is well established, it remains unclear to what degree
microtubules contribute to this process. Herein, we have studied the
contribution of microtubules to cell motility by time-lapse video
microscopy on green flourescence protein-actin- and
tubulin-green fluorescence protein-transfected melanoma cells.
Treatment of cells with either the microtubule-disrupting agent
nocodazole or with the stabilizing agent taxol showed decreased ruffling and lamellipodium formation. However, this was not due to an
intrinsic inability to form ruffles and lamellipodia because both were
restored by stimulation of cells with phorbol 12-myristate 13-acetate
in a Rac-dependent manner, and by stem cell factor in melanoblasts
expressing the receptor tyrosine kinase c-kit. Although ruffling and
lamellipodia were formed without microtubules, the microtubular network
was needed for advancement of the cell body and the subsequent
retraction of the tail. In conclusion, we demonstrate that the
formation of lamellipodia can occur via actin polymerization
independently of microtubules, but that microtubules are required for
cell migration, tail retraction, and modulation of cell adhesion.
Corresponding authors. E-mail address:
ballestr{at}cmu.unige.ch. or Beat.Imhof{at}medecine.unige.ch
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