Pituitary adenylate cyclase-activating polypeptide (PACAP) in zebrafish models of nephrotic syndrome

Eneman, B., Elmonem, M.A., van den Heuvel, L.P., Khodaparast, L., Khodaparast, L., van Geet, C., Freson, K., Levtchenko, E.
PLoS One   12: e0182100 (Journal)
Registered Authors
Embryos, Zebrafish, Morpholino, Platelets, Central nervous system, Edema, Proteinuria, Mouse models
MeSH Terms
  • Animals
  • Blood Platelets/metabolism
  • Ceruloplasmin/metabolism
  • Doxorubicin/toxicity
  • Membrane Proteins/genetics
  • Membrane Proteins/metabolism*
  • Nephrotic Syndrome/etiology
  • Nephrotic Syndrome/genetics
  • Nephrotic Syndrome/metabolism*
  • Peptide Fragments/pharmacology
  • Pericardium/drug effects
  • Pericardium/metabolism
  • Pericardium/pathology
  • Pituitary Adenylate Cyclase-Activating Polypeptide/chemistry
  • Pituitary Adenylate Cyclase-Activating Polypeptide/genetics
  • Pituitary Adenylate Cyclase-Activating Polypeptide/metabolism*
  • Zebrafish
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
28759637 Full text @ PLoS One
Pituitary adenylate cyclase-activating polypeptide (PACAP) is an inhibitor of megakaryopoiesis and platelet function. Recently, PACAP deficiency was observed in children with nephrotic syndrome (NS), associated with increased platelet count and aggregability and increased risk of thrombosis. To further study PACAP deficiency in NS, we used transgenic Tg(cd41:EGFP) zebrafish with GFP-labeled thrombocytes. We generated two models for congenital NS, a morpholino injected model targeting nphs1 (nephrin), which is mutated in the Finnish-type congenital NS. The second model was induced by exposure to the nephrotoxic compound adriamycin. Nephrin RNA expression was quantified and zebrafish embryos were live-screened for proteinuria and pericardial edema as evidence of renal impairment. Protein levels of PACAP and its binding-protein ceruloplasmin were measured and GFP-labeled thrombocytes were quantified. We also evaluated the effects of PACAP morpholino injection and the rescue effects of PACAP-38 peptide in both congenital NS models. Nephrin downregulation and pericardial edema were observed in both nephrin morpholino injected and adriamycin exposed congenital NS models. However, PACAP deficiency was demonstrated only in the adriamycin exposed condition. Ceruloplasmin levels and the number of GFP-labeled thrombocytes remained unchanged in both models. PACAP morpholino injections worsened survival rates and the edema phenotype in both congenital NS models while injection with human PACAP-38 could only rescue the adriamycin exposed model. We hereby report, for the first time, PACAP deficiency in a NS zebrafish model as a consequence of adriamycin exposure. However, distinct from the human congenital NS, both zebrafish models retained normal levels of ceruloplasmin and thrombocytes. We further extend the renoprotective effects of the PACAP-38 peptide against adriamycin toxicity in zebrafish.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes