title: Zebrafish ProVEGF-C expression, proteolytic processing and inhibitory effect of unprocessed ProVEGF-C during fin regeneration.
abstract: BACKGROUND
  In zebrafish, vascular endothelial growth factor-C precursor (proVEGF-C) processing occurs within the dibasic motif HSIIRR(214) suggesting the involvement of one or more basic amino acid-specific proprotein convertases (PCs) in this process. In the present study, we examined zebrafish proVEGF-C expression and processing and the effect of unprocessed proVEGF-C on caudal fin regeneration.
  
  
  RESULTS
  Cell transfection assays revealed that the cleavage of proVEGF-C, mainly mediated by the proprotein convertases Furin and PC5 and to a less degree by PACE4 and PC7, is abolished by PCs inhibitors or by mutation of its cleavage site (HSIIRR(214) into HSIISS(214)). In vitro, unprocessed proVEGF-C failed to activate its signaling proteins Akt and ERK and to induce cell proliferation. In vivo, following caudal fin amputation, the induction of VEGF-C, Furin and PC5 expression occurs as early as 2 days post-amputation (dpa) with a maximum levels at 4-7 dpa. Using immunofluorescence staining we localized high expression of VEGF-C and the convertases Furin and PC5 surrounding the apical growth zone of the regenerating fin. While expression of wild-type proVEGF-C in this area had no effect, unprocessed proVEGF-C inhibited fin regeneration.
  
  
  CONCLUSIONS
  Taken together, these data indicate that zebrafish fin regeneration is associated with up-regulation of VEGF-C and the convertases Furin and PC5 and highlight the inhibitory effect of unprocessed proVEGF-C on fin regeneration.
authors.affiliation: INSERM, UMRS940, Equipe Avenir. Institut de Génétique Moléculaire, Hôpital St-Louis, Université Paris 7, Paris, France.
authors.forename: Abdel-Majid
authors.initials: AM
authors.lastname: Khatib
authors.forename: Rachid
authors.initials: R
authors.lastname: Lahlil
authors.forename: Nathalie
authors.initials: N
authors.lastname: Scamuffa
authors.forename: Marie-Andrée
authors.initials: MA
authors.lastname: Akimenko
authors.forename: Sylvain
authors.initials: S
authors.lastname: Ernest
authors.forename: Abdderahim
authors.initials: A
authors.lastname: Lomri
authors.forename: Claude
authors.initials: C
authors.lastname: Lalou
authors.forename: Nabil G
authors.initials: NG
authors.lastname: Seidah
authors.forename: Bruno O
authors.initials: BO
authors.lastname: Villoutreix
authors.forename: Fabien
authors.initials: F
authors.lastname: Calvo
authors.forename: Geraldine
authors.initials: G
authors.lastname: Siegfried
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D015153:Blotting, Western; D049109:Cell Proliferation; D002478:Cells, Cultured; D045683:Furin / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D045702:Proprotein Convertase 5 / Q000235:genetics / Q000378:metabolism; D012038:Regeneration / Q000235:genetics / Q000502:physiology*; D014443:Tyrosine / Q000378:metabolism; D042582:Vascular Endothelial Growth Factor C / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D042582:Vascular Endothelial Growth Factor C; D029961:Zebrafish Proteins; D014443:Tyrosine; D045702:Proprotein Convertase 5; D045683:Furin