RGDS and DGEA-induced [Ca2+]i signalling in human dermal fibroblasts.
A pulse of short peptides, RGDS and DGEA in the millimolar range, immediately elicits in normal human fibroblasts a transient increase of intracellular Ca2+ ([Ca2+]i). In the present study, we show that this [Ca2+]i occurs in an increasing number of cells as a function of peptides concentration. It is specific of each peptide and inhibited at saturating concentration of the peptide in the culture medium. The [Ca2+]i transient depends on signalling pathways slightly different for DGEA and RGDS involving tyrosine kinase(s) and phosphatase(s), phospholipase C, production of inositol-trisphosphate and release of Ca2+ from the cellular stores. GFOGER, the classical collagen binding peptide of alpha1- alpha2- and alpha11-beta1 integrins, in triple helical or denatured form, does not produce any Ca2+ signal. The [Ca2+]i signalling induced by RGDS and DGEA is inhibited by antibodies against beta1 integrin subunit while that mediated by RGDS is also inhibited by antibodies against the alpha3 integrin. Delay in the acquisition of responsiveness is observed during cell adhesion and spreading on a coat of fibronectin for RGDS or collagen for DGEA or on a coat of the specific integrin-inhibiting antibodies but not by seeding cells on GFOGER or laminin-5. This delay is suppressed specifically by collagenase acting on the collagen coat or trypsin on the fibronectin coat. Our results suggest that free integrins and associated focal complexes generate a Ca2+ signal upon recognition of DGEA and RGDS by different cellular pathways.
Mineur
P
P
Lab. Biologie des Tissus Conjonctifs, University of Liège, Belgium. Pierre.Mineur@ulg.ac.be
Guignandon
A
A
Lambert
Ch A
ChA
Amblard
M
M
Lapière
Ch M
ChM
Nusgens
B V
BV
D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects*; D002413:Cations, Divalent / Q000378:metabolism; D002448:Cell Adhesion; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology*; D011485:Protein Binding; D012867:Skin / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D013379:Substrate Specificity
D002413:Cations, Divalent; D024042:Collagen Type I; D005353:Fibronectins; D007797:Laminin; D010455:Peptides; D002118:Calcium