QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

This Article Figures Only Full Text Full Text (PDF) Movie Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ohashi, T. Articles by Erickson, H. P. Search for Related Content PubMed PubMed Citation Articles by Ohashi, T. Articles by Erickson, H. P.

Dual labeling of the fibronectin matrix and actin cytoskeleton with green fluorescent protein variants

¹ Department of Cell Biology, Duke University Medical Center, NC27710, USA
² Department of Biology, Duke University, NC27708, USA

^* Author for correspondence (e-mail: h.erickson{at}cellbio.duke.edu )

Accepted 19 December 2001

We have prepared 3T3 cells doubly labeled to visualize simultaneously the extracellular fibronectin (FN) matrix and intracellular actin cytoskeleton in living cell cultures. We used FN-yellow fluorescent protein (FN-yfp) for the FN matrix, and the actin-binding domain of moesin fused to cyan fluorescent protein (cfp-Moe) to stain actin. Actin filament bundles were clearly seen in the protruding lamellae of the cells. FN matrix assembly appeared to be initiated as small spots of FN at the ends of actin filament bundles. The spots then elongated along the actin filament bundle toward the cell center to form FN fibrils. The end of the fibril towards the cell edge appeared immobile, and probably attached to the substrate, whereas the end toward the cell center frequently showed movements, suggesting attachment to the cell. Combining our data with the observations of Pankov et al. we suggest that fibrils grow by stretching this mobile end toward the cell center while adding new FN molecules at the end and along the entire lenght. When the cell culture was treated with cytochalasin to disrupt the actin cytoskeleton, some fibrils contracted substantially, suggesting that the segment attached primarily to the cell surface is stretched.

Key words: Fibronectin, Actin, Matrix assembly, Moesin, Green fluorescent protein

This article has been cited by other articles:

				C. C. Feral, A. Zijlstra, E. Tkachenko, G. Prager, M. L. Gardel, M. Slepak, and M. H. Ginsberg CD98hc (SLC3A2) participates in fibronectin matrix assembly by mediating integrin signaling J. Cell Biol., August 9, 2007; 178(4): 701 - 711. [Abstract] [Full Text] [PDF]

				P. Sivakumar, A. Czirok, B. J. Rongish, V. P. Divakara, Y.-P. Wang, and S. L. Dallas New insights into extracellular matrix assembly and reorganization from dynamic imaging of extracellular matrix proteins in living osteoblasts J. Cell Sci., April 1, 2006; 119(7): 1350 - 1360. [Abstract] [Full Text] [PDF]

				L. H. Romer, K. G. Birukov, and J. G.N. Garcia Focal Adhesions: Paradigm for a Signaling Nexus Circ. Res., March 17, 2006; 98(5): 606 - 616. [Abstract] [Full Text] [PDF]

				T. Ohashi and H. P. Erickson Domain Unfolding Plays a Role in Superfibronectin Formation J. Biol. Chem., November 25, 2005; 280(47): 39143 - 39151. [Abstract] [Full Text] [PDF]

				E. E. Robinson, R. A. Foty, and S. A. Corbett Fibronectin Matrix Assembly Regulates {alpha}5{beta}1-mediated Cell Cohesion Mol. Biol. Cell, March 1, 2004; 15(3): 973 - 981. [Abstract] [Full Text] [PDF]

				I. Wierzbicka-Patynowski and J. E. Schwarzbauer The ins and outs of fibronectin matrix assembly J. Cell Sci., August 15, 2003; 116(16): 3269 - 3276. [Abstract] [Full Text] [PDF]