title: Jararhagin, a snake venom metalloproteinase-disintegrin, stimulates epithelial cell migration in an in vitro restitution model.
abstract: The snake venom metalloproteinase-disintegrin jararhagin (JG) has no chemotactic activity but stimulates the migration of neutrophils in vivo through a mechanism still unclear. In this study we investigated the effects of jararhagin on epithelial cell adhesion and migration in vitro. F-actin arrangement and the distribution of laminin, fibronectin, several integrins and phosphorylated Focal Adhesion Kinase (FAK) were studied using rhodamine-phalloidin and immunofluorescence. Maximum stimulation of migration (about 100%) was obtained with 5 microg/ml JG, with about 38% inhibition of cellular adhesion. In migratory cells the toxin stimulated the formation of filopodia, lamellipodia and stress fibers. The pericellular fibronectin matrix was lost in migrating cells, while laminin was less affected. The toxin stimulated FAK phosphorylation and the recruitment of alphav-containing integrins to focal contacts, whereas integrins containing the alpha2 subunit were reduced in these junctions. Inactivation of the toxin with 1,10 phenanthroline showed that the catalytic activity is important for the effect of jararhagin on cell migration, FAK phosphorylation and for the recruitment of alphav, but not as much for the anti-adhesive effect. In conclusion, jararhagin stimulates the migration of epithelial cells in vitro through a mechanism that involves its proteolytic activity, qualitative changes in cellular adhesion and the formation of actin-rich cellular processes.
authors.affiliation: Department of Developmental and Cell Biology, Institute of Biomedical Sciences, University of São Paulo, ICB/USP, Av. Prof. Lineu Prestes 1524, São Paulo, SP CEP 05508-000, Brazil.
authors.forename: Erica Pereira
authors.initials: EP
authors.lastname: Costa
authors.forename: Marinilce Fagundes
authors.initials: MF
authors.lastname: Santos
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D017837:Bothrops; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D003435:Crotalid Venoms / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D005353:Fibronectins / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases; D039421:Integrin alpha2 / Q000378:metabolism; D039564:Integrin alphaV / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D008666:Metalloendopeptidases / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D000097146:Bothrops jararaca Venom
chemical_list: D000199:Actins; D003435:Crotalid Venoms; D005353:Fibronectins; D051416:Focal Adhesion Protein-Tyrosine Kinases; D039421:Integrin alpha2; D039564:Integrin alphaV; D007797:Laminin; D008666:Metalloendopeptidases; D011505:Protein-Tyrosine Kinases; D000097146:Bothrops jararaca Venom