Extracellular matrix fibers containing fibronectin and basement membrane heparan sulfate proteoglycan coalign with focal contacts and microfilament bundles in stationary fibroblasts

Experimental Cell Research
I I SingerJ R Hassell

Abstract

Double-label immunofluorescence microscopy and immunoelectron microscopy were performed on stationary cultures of Nil 8 fibroblasts to determine if fibronectin and basement membrane heparan sulfate proteoglycans play coordinated roles in cell-to-substrate adhesion. Relationships between subcellular matrix fibers containing fibronectin plus proteoglycan, and focal contacts associated with microfilament bundles, were studied simultaneously using interference reflection microscopy, differential interference contrast microscopy, and immunofluorescence microscopy. Cells maintained in 0.3% FBS were doubly stained with monospecific anti-fibronectin IgG and antibodies against a basement membrane proteoglycan purified from the EHS (Engelbreth-Holm-Swarm) tumor. Coincident patterns of fibronectin and proteoglycan-containing fibers were found to codistribute with focal contacts and microfilament bundles in both early (6-h) and late (24-h) cultures. The early cells showed doubly-stained fibers colinear with substrate adhesion sites in 43% of the sample, while 100% of the later cells exhibited these coaligned matrix-cytoskeletal attachment complexes. Immunoelectron microscopy showed that both of these antigens were situated in the same type...Continue Reading

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Citations

Jan 1, 1991·Mammalian Genome : Official Journal of the International Mammalian Genome Society·S ChakravartiJ R Hassell
Jan 24, 2002·The Journal of Biological Chemistry·Satoshi HirohataSuneel S Apte
Jan 1, 1989·Arteriosclerosis : an Official Journal of the American Heart Association, Inc·T N Wight
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Jan 20, 2005·The Journal of Biological Chemistry·Kerstin TiedemannDieter P Reinhardt

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