Cell-matrix interactions induce tyrosine phosphorylation of MAP kinases ERK1 and ERK2 and PLCgamma-1 in two-dimensional and three-dimensional cultures of human fibroblasts

Exp Cell Res. 1997 Aug 25;235(1):22-7. doi: 10.1006/excr.1997.3640.


Using immunoprecipitation and phosphotyrosine detection by Western blotting, intracellular signaling intermediates were analyzed in human primary dermal fibroblasts, either seeded as monolayers on collagen I coats (2D) or seeded within three-dimensional collagen I lattices (3D). Previous results demonstrated that integrin activation in these systems resulted in a cascade of protein tyrosine phosphorylation, including focal adhesion kinase (D. Roeckel and T. Krieg, 1994, Exp. Cell Res. 211, 42-48). Further downstream signaling events are now shown to include coordinate activation of ERK1 and ERK2 at 2 h after cell-collagen contact, irrespective of 2D or 3D culture conditions. Applying U-73122, an inhibitor of PLC, inhibits collagen lattice contraction in a dose-dependent fashion. Immunoprecipitation identified the isoform PLCgamma-1 as playing a role as signaling intermediate in fibroblast-collagen interactions. PLCgamma-1 becomes phosphorylated within 10 min after culture initiation and declines after 2 h. So far, no qualitative differences in signaling intermediates between 2D and 3D cultures have been identified.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
  • Cell Adhesion
  • Cell Culture Techniques / methods
  • Collagen
  • Enzyme Activation
  • Enzyme Inhibitors / pharmacology
  • Estrenes / pharmacology
  • Extracellular Matrix / physiology*
  • Fibroblasts / cytology
  • Fibroblasts / drug effects
  • Fibroblasts / physiology
  • Humans
  • Isoenzymes / antagonists & inhibitors
  • Isoenzymes / metabolism*
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases*
  • Phospholipase C gamma
  • Phosphorylation
  • Phosphotyrosine / metabolism*
  • Pyrrolidinones / pharmacology
  • Skin / cytology*
  • Skin Physiological Phenomena*
  • Type C Phospholipases / antagonists & inhibitors
  • Type C Phospholipases / metabolism*


  • Enzyme Inhibitors
  • Estrenes
  • Isoenzymes
  • Pyrrolidinones
  • 1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione
  • Phosphotyrosine
  • Collagen
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases
  • Type C Phospholipases
  • Phospholipase C gamma