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PMID: 10051610 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Dynamics and elasticity of the fibronectin matrix in living cell culture visualized by fibronectin-green fluorescent protein.

Ohashi T, Kiehart DP, Erickson HP

Abstract

Fibronectin (FN) forms the primitive fibrillar matrix in both embryos and healing wounds. To study the matrix in living cell cultures, we have constructed a cell line that secretes FN molecules chimeric with green fluorescent protein. These FN-green fluorescent protein molecules were assembled into a typical matrix that was easily visualized by fluorescence over periods of several hours. FN fibrils remained mostly straight, and they were seen to extend and contract to accommodate movements of the cells, indicating that they are elastic. When fibrils were broken or detached from cells, they contracted to less than one-fourth of their extended length, demonstrating that they are highly stretched in the living culture. Previous work from other laboratories has suggested that cryptic sites for FN assembly may be exposed by tension on FN. Our results show directly that FN matrix fibrils are not only under tension but are also highly stretched. This stretched state of FN is an obvious candidate for exposing the cryptic assembly sites.

MeSH Terms
Amino Acid Sequence Animals CHO Cells Cell Membrane/physiology,ultrastructure Cells, Cultured Cricetinae Elasticity Extracellular Matrix/physiology,ultrastructure Fibronectins/chemistry,genetics,physiology Green Fluorescent Proteins Kinetics Luminescent Proteins/analysis,chemistry,genetics Mutagenesis, Insertional Polymerase Chain Reaction Recombinant Fusion Proteins/analysis,chemistry,metabolism Time Factors Transfection
Chemicals
Fibronectins Luminescent Proteins Recombinant Fusion Proteins Green Fluorescent Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ohashi T
Department of Cell Biology, Duke University Medical Center, Durham, NC 27710-3011, USA.
Kiehart D P
Erickson H P
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1999-03-02
Pages
2153-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC26752
Subset
IM
Grants
NCI NIH HHS · CA07456 · United States
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