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Loss of caveolin-1 polarity impedes endothelial cell polarization and directional movement.

Authors
Beardsley-A; Fang-K; Mertz-H; Castranova-V; Friend-S; Liu-J
Source
J Biol Chem 2005 Feb; 280(5):3541-3547
NIOSHTIC No.
20026079
Abstract
The ability of a cell to move requires the asymmetrical organization of cellular activities. To investigate polarized cellular activity in moving endothelial cells, human endothelial cells were incubated in a Dunn chamber to allow migration toward vascular endothelial growth factor. Immunofluorescent staining with a specific antibody against caveolin-1 revealed that caveolin-1 was concentrated at the rear of moving cells. Similarly, monolayer scraping to induce random cell walk resulted in relocation of caveolin-1 to the cell rear. These results suggest that posterior polarization of caveolin-1 is a common feature both for chemotaxis and chemokinesis. Dual immunofluorescent labeling showed that, during cell spreading, caveolin-1 was compacted in the cell center and excluded from nascent focal contacts along the circular lamellipodium, as revealed by integrin beta1 and FAK staining. When cells were migrating, integrin beta1 and FAK appeared at polarized lamellipodia, whereas caveolin-1 was found at the posterior of moving cells. Notably, wherever caveolin-1 was polarized, there was a conspicuous absence of lamellipod protrusion. Transmission electron microscopy showed that caveolae, similar to their marker caveolin-1, were located at the cell center during cell spreading or at the cell rear during cell migration. In contrast to its unphosphorylated form, tyrosine-phosphorylated caveolin-1, upon fibronectin stimulation, was associated with the focal complex molecule phosphopaxillin along the lamellipodia of moving cells. Thus, unphosphorylated and phosphorylated caveolin-1 were located at opposite poles during cell migration. Importantly, loss of caveolin-1 polarity by targeted down-regulation of the protein prevented cell polarization and directional movement. Our present results suggest a potential role of caveolin polarity in lamellipod extension and cell migration.
Keywords
Cell-function; Cell-migration; Cellular-function; Cell-cultures; Kinetics
Contact
Jun Liu, Deptartment of Physiology and Pharmacology, Health Science Center, West Virginia University, P. O. Box 9229, Morgantown, WV 26506-9229
CODEN
JBCHA3
Publication Date
20050204
Document Type
Journal Article
Email Address
junliu@hsc.wvu.edu
Fiscal Year
2005
NTIS Accession No.
NTIS Price
Issue of Publication
5
ISSN
0021-9258
NIOSH Division
HELD
Source Name
Journal of Biological Chemistry
State
WV
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