title: Activation of the MAPKs ERK1/2 by cell swelling in turbot hepatocytes.
abstract: BACKGROUND
  Activation of MAPKs (mitogen-activated protein kinases), in particular ERK1/2 (extracellular-signal-regulated kinase 1/2), has been reported to take place in a large variety of cell types after hypo-osmotic cell swelling. Depending on cell type, ERK1/2 phosphorylation can then serve or not the RVD (regulatory volume decrease) process. The present study investigates ERK1/2 activation after aniso-osmotic stimulations in turbot hepatocytes and the potential link between phosphorylation of these proteins and RVD.
  
  
  RESULTS
  In turbot hepatocytes, Western-blot analysis shows that a hypo-osmotic shock from 320 to 240 mOsm kg(-1) induced a rapid increase in ERK1/2 phosphorylation, whereas a hyper-osmotic shock from 320 to 400 mOsm kg(-1) induced no significant change in the phosphorylation of these proteins. The hypo-osmotic-induced ERK1/2 phosphorylation was significantly prevented when hypo-osmotic shock was performed in the presence of the specific MEK (MAPK/ERK kinase) inhibitor PD98059 (100 microM). In these conditions, the RVD process was not altered, suggesting that ERK1/2 did not participate in this process in turbot hepatocytes. Moreover, the hypo-osmotic-induced activation of ERK1/2 was significantly prevented by breakdown of extracellular ATP by apyrase (10 units ml(-1)), by inhibition of purinergic P2 receptors by suramin (100 microM) or by calcium depletion using EGTA (1 mM) and thapsigargin (1 microM).
  
  
  CONCLUSIONS
  In turbot hepatocytes, hypo-osmotic swelling but not hyper-osmotic shrinkage induced the activation of ERK1/2. However, these proteins do not seem to be involved in the RVD process. Their hypo-osmotic-induced activation is partially due to cascades of signalling events triggered by the binding of released ATP on purinergic P2 receptors and requires the presence of calcium.
authors.affiliation: Laboratoire ORPHY, Université Européenne de Bretagne, Université de Brest, Brest, France. Audrey.Fouchs@univ-brest.fr
authors.forename: Audrey
authors.initials: A
authors.lastname: Fouchs
authors.forename: Hélène
authors.initials: H
authors.lastname: Ollivier
authors.forename: Christophe
authors.initials: C
authors.lastname: Haond
authors.forename: Stella
authors.initials: S
authors.lastname: Roy
authors.forename: Patrick
authors.initials: P
authors.lastname: Calvès
authors.forename: Karine
authors.initials: K
authors.lastname: Pichavant-Rafini
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000502:physiology; D048429:Cell Size; D004533:Egtazic Acid / Q000494:pharmacology; D004789:Enzyme Activation; D005412:Flatfishes; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology*; D007038:Hypotonic Solutions / Q000494:pharmacology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D009997:Osmotic Pressure; D010766:Phosphorylation; D018048:Receptors, Purinergic P2 / Q000378:metabolism; D019284:Thapsigargin / Q000494:pharmacology
chemical_list: D007038:Hypotonic Solutions; D018048:Receptors, Purinergic P2; D004533:Egtazic Acid; D019284:Thapsigargin; D000255:Adenosine Triphosphate; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D002118:Calcium