Exogenous tenascin inhibits mesodermal cell migration during amphibian gastrulation.
We have used amphibian gastrulation as a model system to study the action of the extracellular matrix (ECM) glycoprotein tenascin on mesodermal cell migration. Tenascin function was assayed in vitro during spreading of isolated cells from the dorsal marginal zone (DMZ) and during cell migration from DMZ explants. Plastic coated with bovine fibronectin or gastrula ECM was used as a substratum. In both cases, tenascin added to the medium inhibited spreading and migration of mesodermal cells. In addition, a substratum coated with a mixture of fibronectin and tenascin was found to prevent mesodermal cell migration. Tenascin was also microinjected into the blastocoel cavity of living embryos at the late blastula stage. This led to a complete arrest of gastrulation in more than 80% of the cases. Scanning electron microscopy of fractures from arrested gastrulae showed that mesodermal cell migration was blocked. Similar injection experiments carried out at the middle gastrula stage demonstrated that tenascin is able to inhibit cell migration after cells have already contacted the ECM. Mesodermal cell migration in the presence of tenascin could be restored in vitro and in vivo by the monoclonal antibody mAb Tn68 which is known to mask a cell binding site of the molecule. Finally, tenascin microinjected into the blastocoel of blastula or gastrula stage embryos bound within 15 min to the ECM fibrils at all the stages studied. Our results show that exogenous tenascin can be incorporated into embryonic ECM and interferes in vivo with the interactions of cells with a fibronectin-rich matrix.
Laboratoire de Biologie Expérimentale, U.A. 1135 CNRS, Université Pierre et Marie Curie, Paris, France.
J F
JF
Riou
D L
DL
Shi
M
M
Chiquet
J C
JC
Boucaut
D000818:Animals; D000911:Antibodies, Monoclonal; D002448:Cell Adhesion; D015816:Cell Adhesion Molecules, Neuronal / Q000276:immunology / Q000494:pharmacology* / Q000502:physiology; D002465:Cell Movement / Q000187:drug effects*; D004625:Embryo, Nonmammalian / Q000502:physiology*; D005109:Extracellular Matrix / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D005775:Gastrula / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D008648:Mesoderm / Q000187:drug effects / Q000502:physiology*; D008855:Microscopy, Electron, Scanning; D010999:Pleurodeles; D019063:Tenascin
D000911:Antibodies, Monoclonal; D015816:Cell Adhesion Molecules, Neuronal; D019063:Tenascin