Submitted July 18, 2008
Accepted February 24, 2009
Zebrafish mutants in the von Hippel-Lindau (VHL) tumor suppressor display a hypoxic response and recapitulate key aspects of Chuvash polycythemia
Ellen van Rooijen, Emile E. Voest, Ive Logister, Jeroen Korving, Thorsten Schwerte, Stefan Schulte-Merker, Rachel H. Giles, and Fredericus J. van Eeden*
Department of Medical Oncology, University Medical Center, Utrecht, Netherlands
Hubrecht Institute - KNAW & University Medical Center, Utrecht, Netherlands
Institute of Zoology, University of Innsbruck, Innsbruck, Austria
* Corresponding author; email: f.j.vaneeden{at}sheffield.ac.uk.
We have generated two 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 upregulation of hypoxia-induced genes by 1 day post-fertilization and a severe hyperventilation and cardiophysiological response. vhl mutants develop polycythemia with concomitantly increased epo/epor mRNA levels and Epo 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+ HSCs and circulating erythroid precursors. Chemical activation of Hif-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.
