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Vol. 11, Issue 7, 2471-2483, July 2000
and
*Department of Cell Biology and Institute for Childhood and
Neglected Diseases, The Scripps Research Institute, La Jolla,
California 92037; and To test how cell-cell contacts regulate microtubule (MT) and actin
cytoskeletal dynamics, we examined dynamics in cells that were
contacted on all sides with neighboring cells in an epithelial cell
sheet that was undergoing migration as a wound-healing response. Dynamics were recorded using time-lapse digital fluorescence microscopy of microinjected, labeled tubulin and actin. In fully contacted cells,
most MT plus ends were quiescent; exhibiting only brief excursions of
growth and shortening and spending 87.4% of their time in pause. This
contrasts MTs in the lamella of migrating cells at the noncontacted
leading edge of the sheet in which MTs exhibit dynamic
instability. In the contacted rear and side edges of these
migrating cells, a majority of MTs were also quiescent, indicating that
cell-cell contacts may locally regulate MT dynamics. Using
photoactivation of fluorescence techniques to mark MTs, we found that
MTs in fully contacted cells did not undergo retrograde flow toward the
cell center, such as occurs at the leading edge of motile cells.
Time-lapse fluorescent speckle microscopy of fluorescently labeled
actin in fully contacted cells revealed that actin did not flow
rearward as occurs in the leading edge lamella of migrating cells. To
determine if MTs were required for the maintenance of cell-cell
contacts, cells were treated with nocodazole to inhibit MTs. After 1-2
h in either 10 µM or 100 nM nocodazole, breakage of cell-cell
contacts occurred, indicating that MT growth is required for
maintenance of cell-cell contacts. Analysis of fixed cells indicated
that during nocodazole treatment, actin became reduced in adherens
junctions, and junction proteins Department of Biology, University
of North Carolina, Chapel Hill, North Carolina 27599
- and 
-catenin were lost from
adherens junctions as cell-cell contacts were broken. These results
indicate that a MT plus end capping protein is regulated by cell-cell
contact, and in turn, that MT growth regulates the maintenance of
adherens junctions contacts in epithelia.
Online version of this article contains video
material to accompany Figures 1-4. Online version is available at
www.molbiolcell.org
Corresponding author. E-mail address:
waterman{at}scripps.edu.
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