Integrin-like substrate adhesion in RTG-2 cells, a fibroblastic cell line derived from rainbow trout.
Fluorometric cell attachment assays together with competitive inhibitors of adhesion were used to probe for the presence of integrins, a diverse family of heterodimeric cell-surface glycoproteins involved in cell-cell and cell-extracellular matrix adhesion, in the fibroblastic rainbow trout cell line, RTG-2. The adhesive properties of this cell line were evaluated. RTG-2 cells adhered poorly to TC plastic in the absence of serum but as little as 2.5% fetal bovine serum allowed over 75% of the cells to attach after 5 h. Surfaces coated with the extracellular matrix proteins collagen I, collagen IV, fibrin, fibrinogen, or fibronectin were able to support attachment of RTG-2 cells. Adhesion of RTG-2 cells to fibronectin varied linearly with fibronectin coating densities in the range 0 to 65 ng/mm(2). Oligopeptides containing the sequence Arg-Gly-Asp (RGD) caused dose-dependent inhibition of adhesion to microtiter plates coated with fibrin, fibrinogen, and fibronectin, whereas attachment to collagen I and collagen IV was less severely affected. In all cases, peptides containing Arg-Gly-Glu (RGE) or Asp-Gly-Arg (DGR) sequences caused no reduction of cell attachment. Since many integrins mediate adhesion by binding to RGD sequences in their target ligands, these results suggest the presence of integrin-like adhesion molecules on the surface of RTG-2 cells.
Reinhart
B
B
Department of Biology, Wilfrid Laurier University, Waterloo, Ontario, N2L 3C5, Canada.
Lee
L E J
LE
D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D015815:Cell Adhesion Molecules; D002460:Cell Line; D016895:Culture Media, Serum-Free / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000502:physiology; D005347:Fibroblasts / Q000378:metabolism*; D016023:Integrins / Q000502:physiology*; D009842:Oligopeptides / Q000737:chemistry / Q000502:physiology; D011485:Protein Binding; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D014337:Trout
D015815:Cell Adhesion Molecules; D016895:Culture Media, Serum-Free; D016326:Extracellular Matrix Proteins; D016023:Integrins; D009842:Oligopeptides; D011500:Protein Synthesis Inhibitors; D003513:Cycloheximide