title: Zebrafish mutants in the von Hippel-Lindau tumor suppressor display a hypoxic response and recapitulate key aspects of Chuvash polycythemia.
abstract: We have generated 2 zebrafish lines carrying inactivating germline mutations in the von Hippel-Lindau (VHL) tumor suppressor gene ortholog vhl. Mutant embryos display a general systemic hypoxic response, including the up-regulation of hypoxia-induced genes by 1 day after fertilization and a severe hyperventilation and cardiophysiologic response. The vhl mutants develop polycythemia with concomitantly increased epo/epor mRNA levels and erythropoietin signaling. In situ hybridizations reveal global up-regulation of both red and white hematopoietic lineages. Hematopoietic tissues are highly proliferative, with enlarged populations of c-myb(+) hematopoietic stem cells and circulating erythroid precursors. Chemical activation of hypoxia-inducible factor signaling recapitulated aspects of the vhl(-/-) phenotype. Furthermore, microarray expression analysis confirms the hypoxic response and hematopoietic phenotype observed in vhl(-/-) embryos. We conclude that VHL participates in regulating hematopoiesis and erythroid differentiation. Injections with human VHLp30 and R200W mutant mRNA demonstrate functional conservation of VHL between mammals and zebrafish at the amino acid level, indicating that vhl mutants are a powerful new tool to study genotype-phenotype correlations in human disease. Zebrafish vhl mutants are the first congenital embryonic viable systemic vertebrate animal model for VHL, representing the most accurate model for VHL-associated polycythemia to date. They will contribute to our understanding of hypoxic signaling, hematopoiesis, and VHL-associated disease progression.
authors.affiliation: Department of Medical Oncology, University Medical Center, Utrecht, The Netherlands.
authors.forename: Ellen
authors.initials: E
authors.lastname: van Rooijen
authors.forename: Emile E
authors.initials: EE
authors.lastname: Voest
authors.forename: Ive
authors.initials: I
authors.lastname: Logister
authors.forename: Jeroen
authors.initials: J
authors.lastname: Korving
authors.forename: Thorsten
authors.initials: T
authors.lastname: Schwerte
authors.forename: Stefan
authors.initials: S
authors.lastname: Schulte-Merker
authors.forename: Rachel H
authors.initials: RH
authors.lastname: Giles
authors.forename: Fredericus J
authors.initials: FJ
authors.lastname: van Eeden
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis / Q000235:genetics; D019070:Cell Lineage; D017124:Conserved Sequence; D004195:Disease Models, Animal; D055786:Gene Knockout Techniques; D018095:Germ-Line Mutation; D006410:Hematopoiesis / Q000235:genetics; D006801:Humans; D000860:Hypoxia / Q000235:genetics* / Q000503:physiopathology; D008969:Molecular Sequence Data; D017354:Point Mutation; D011086:Polycythemia / Q000235:genetics* / Q000503:physiopathology; D012333:RNA, Messenger / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000502:physiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D026801:Synteny; D025521:Tumor Suppressor Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D025521:Tumor Suppressor Proteins; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D029961:Zebrafish Proteins; C542158:Vhl protein, zebrafish; C497446:VHL protein, human