This Article Full Text Full Text (PDF) Supplemental Data for this article All Versions of this Article: jhc.2009.953802v1 57/11/1041    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services 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 Google Scholar Google Scholar Articles by Evanko, S. P. Articles by Wight, T. N. Search for Related Content PubMed PubMed Citation Articles by Evanko, S. P. Articles by Wight, T. N. Social Bookmarking                         What's this?

Organization of Hyaluronan and Versican in the Extracellular Matrix of Human Fibroblasts Treated With the Viral Mimetic Poly I:C

Stephen P. Evanko, Susan Potter-Perigo, Pamela Y. Johnson and Thomas N. Wight

Hope Heart Program, Benaroya Research Institute at Virginia Mason, Seattle, Washington

Correspondence to: Thomas N. Wight, PhD, Director, Hope Heart Program, Benaroya Research Institute at Virginia Mason, 1201 Ninth Avenue, Seattle, WA 98101. E-mail: twight{at}benaroyaresearch.org

We have examined structural details of hyaluronan- and versican-rich pericellular matrices in human lung fibroblasts, as well as fixation effects after treatment with the viral mimetic, poly I:C. Lateral aggregation of hyaluronan chains was promoted by acid-ethanol-formalin fixation compared with a network appearance with formalin alone. However, hyaluronidase-sensitive cable structures were seen in live cells, suggesting that they are not a fixation artifact. With all fixatives, versican and hyaluronan probes bound alternately along strands extending from the plasma membrane. However, a yellow colocalization signal required aggregation/overlap of several hyaluronan/versican strands and was more pronounced after acid-ethanol-formalin fixation. In addition to the main cell surface, hyaluronan and versican were also associated with fine actin-positive membrane protrusions, retraction fibers, and surface blebs. After wounding plus treatment with poly I:C, cells displayed larger hyaluronan coats and cable-like structures, as well as more membrane protrusions. However, treated cells did not migrate and had increased stress fibers compared with control wounded cells. Deposition of hyaluronan into cable-like structures in response to poly I:C was diminished but still apparent following actin filament disruption with cytochalasin D, suggesting that the protrusions only partially facilitate cable formation. As seen by scanning electron microscopy, the membrane protrusions may participate in poly I:C–induced binding of monocytes to hyaluronan- and versican-rich matrices. These results suggest that poly I:C–induced hyaluronan- and versican-rich cable structures are not deposited during migration, and that cellular protrusions partially contribute to hyaluronan cable formation. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. (J Histochem Cytochem 57:1041–1060, 2009)

Key Words: hyaluronan • versican • extracellular matrix • pericellular space • monocytes • migration • poly I:C

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact

The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 2009