title: Evidence for cell surface extracellular matrix binding proteins in Hydra vulgaris.
abstract: The present study was designed to identify and functionally characterize potential cell surface extracellular matrix binding proteins in Hydra vulgaris. Using [3H]-laminin as a probe, radioreceptor analysis of a dissociated mixed hydra cell preparation indicated that the average number of laminin binding sites per cell was about 10,000 with a dissociation constant of 1.49 nM. These binding sites could be displaced with unlabelled laminin in a dose-dependent manner and with high concentrations (500 nM) of unlabelled fibronectin. No displacement with type-IV collagen and type-I collagen was observed. Immunoscreening studies with a battery of antibodies raised to mammalian extracellular matrix (ECM) binding proteins indicated potential cell surface binding sites for the anti-beta 1 integrin monoclonal antibody, mAb JG22. Cell adhesion studies indicated that mAb JG22 blocked binding of hydra cells to laminin, but did not affect their binding to fibronectin, type-IV collagen, or type-I collagen. Light and electron microscopic immunocytochemical studies indicated that mAb JG22 localized to the basal plasma membrane of ectodermal and endodermal epithelial cells. Immunoprecipitation studies identified tow major bands with masses of about 196 kDa and 150 kDa under reducing conditions, and two bands with masses of > 200 kDa under non-reducing conditions. Functional studies indicated that mAb JG22 could reversibly block morphogenesis of hydra cell aggregates, and could block in vivo interstitial cell migration in hydra grafts. These observations indicate that hydra has cell surface binding sites for ECM components which are functionally important during development of this simple Cnidarian.
authors.affiliation: Department of Anatomy & Cell Biology, University of Kansas Medical Center, Kansas City 66160.
authors.forename: A
authors.initials: A
authors.lastname: Ağbaş
authors.forename: M P
authors.initials: MP
authors.lastname: Sarras
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D003094:Collagen / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000502:physiology / Q000648:ultrastructure; D005353:Fibronectins / Q000494:pharmacology; D006801:Humans; D006829:Hydra / Q000166:cytology / Q000502:physiology*; D015151:Immunoblotting; D007074:Immunoglobulin G; D007700:Kinetics; D007797:Laminin / Q000378:metabolism*; D011869:Radioligand Assay; D017474:Receptors, Laminin / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D014316:Tritium
chemical_list: D000911:Antibodies, Monoclonal; D005353:Fibronectins; D007074:Immunoglobulin G; D007797:Laminin; D017474:Receptors, Laminin; D014316:Tritium; D003094:Collagen