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Cancer Cell death Cell cycle Cytoskeleton Exo/endocytosis Differentiation Division Organelles Signalling Stem cells Trafficking
Cell Biology International (2002) 26, 131–144 (Printed in Great Britain)
PERTURBATION OF β1 INTEGRIN FUNCTION USING ANTI-SENSE OR FUNCTION-BLOCKING ANTIBODIES ON CORNEAL CELLS GROWN ON FIBRONECTIN AND TENASCIN
Kathleen J. Doanea,f1, Raka Bhattacharyaa,f2 and Jeff Marchantb
aDepartment of Anatomy, Northeastern Ohio Universities College of Medicine, 4209 State Route 44, P.O. Box 95, Rootstown, OH 44272-0095, U.S.A.
bDepartment of Anatomy and Cellular Biology, Tufts University School of Medicine, U.S.A.


Abstract

During corneal development, neural crest derivatives from the periocular mesenchyme migrate into the cornea and differentiate into corneal fibroblasts. During this time, these cells interact with a variety of extracellular matrices for proper orientation and development. In the present studies, we have examined the interaction of β1 integrins on periocular mesenchyme cells (POM) and corneal fibroblasts (CF) with fibronectin and tenascin by perturbing the function of this integrin. POM and CF attached and spread to a much greater extent on fibronectin than on tenascin. An antibody against β1 integrin, CSAT, decreased spreading and attachment, and resulted in a lack of immuno-detectable β1 integrin in focal adhesions on fibronectin; few β1 positive focal adhesions were observed in cells grown on tenascin. An anti-sense retroviral construct decreased endogenous levels of β1 integrin protein, and caused decreased attachment and spreading as well as sparse, disorganized focal adhesions. These data indicate that in vitro, both POM and CF have β1 integrins that interact with fibronectin and allow them to attach and spread, while tenascin is anti-adhesive. Further studies using both of these experimental paradigms will clarify whether these interactions also occur in vivo.


Key words: cornea, fibronectin, tenascin, integrin, CSAT, anti-sense retroviral construct.

f1To whom correspondence and reprint requests should be addressed: Dr Kathleen J. Doane, Fax: (330) 325-5913. E-mail:kd@neoucom.edu

f2Currently at University of South Florida/Veterans Administration Hospital.


Received 15 August 2000; accepted 24 July 2001

doi:10.1006/cbir.2001.0818


ISSN Print: 1065-6995
ISSN Electronic: 1095-8355
Published by Portland Press Limited on behalf of the International Federation for Cell Biology (IFCB)