STRESS-RELAXATION OF FIBROBLASTS IN COLLAGEN MATRICES TRIGGERS ECTOCYTOSIS OF PLASMA-MEMBRANE VESICLES CONTAINING ACTIN, ANNEXIN-II AND ANNEXIN-VI, AND BETA-1 INTEGRIN RECEPTORS
Tl. Lee et al., STRESS-RELAXATION OF FIBROBLASTS IN COLLAGEN MATRICES TRIGGERS ECTOCYTOSIS OF PLASMA-MEMBRANE VESICLES CONTAINING ACTIN, ANNEXIN-II AND ANNEXIN-VI, AND BETA-1 INTEGRIN RECEPTORS, Journal of Cell Science, 105, 1993, pp. 167-177
To learn about the effects of tension on fibroblast function, we have
been studying initial cellular responses to stress-relaxation. Human f
oreskin fibroblasts were cultured in anchored collagen matrices for 2
days, during which time mechanical stress developed. Subsequently, the
matrices were dislodged; thereby allowing stress to dissipate. Within
5 min after initiating stress-relaxation, fibroblasts retracted their
pseudopodia. At this time, we observed the disappearance of cellular
stress fibers and the formation of actin clusters along the cell margi
ns. The actin was found to be located inside 200 nm diameter vesicles
that were budding from the cell surface. Vesicles isolated from the ma
trix after stress-relaxation contained prominent 24 kDa, 36 kDa (doubl
et), 45 kDa, and 135 kDa polypeptides. The 45 kDa polypeptide was the
major component in the Triton-insoluble vesicle fraction and appeared
to be actin. The 36 kDa (doublet) polypeptide, which was found predomi
nantly in the Triton-soluble vesicle fraction, was identified as annex
in II. Vesicles also contained annexin VI and beta1 integrin receptors
but not tubulin, vimentin, vinculin or annexin I. The results suggest
that stress-relaxation of fibroblasts induces a novel ectocytotic pro
cess involving transient budding of intact, plasma membrane vesicles f
rom the cell cortex. On the basis of their morphological and biochemic
al features, these vesicles may be analogous to the 'matrix vesicles'
released by chondrocytes and could play a role in extracellular matrix
remodeling after wound contraction.