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Blood, Vol. 113, Issue 25, 6449-6460, June 18, 2009
Zebrafish mutants in the von Hippel-Lindau tumor suppressor display a hypoxic response and recapitulate key aspects of Chuvash polycythemia
Blood van Rooijen et al.
113: 6449
Supplemental materials for: van Rooijen et al
Files in this Data Supplement:
- Document 1. Detailed materials and methods (PDF, 48.7 KB)
- Table S1. vhl−∕− microarray expression profile (XLS, 816 KB)
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44k Agilent microarray expression profile of 7 dpf vhl mutants compared to age-matched siblings (P ≤ 0.01).
- Figure S1. Phylogenetic tree of VHL homologues (JPG, 62.8 KB)
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An unrooted phylogenetic tree was constructed (http://www.ebi.ac.uk/Tools/clustalw2/index.html), which shows zebrafish Vhl to cluster with VHL homologs from fish to mammals, confirming its orthology. The putative Vhl-like protein is only present zebrafish, minnow, rainbow trout, and dogfish, which cluster together in a separate group from other fishes and higher vertebrates, suggesting this gene could be a fish-specific duplication. Human VHL-like protein (VLP), which behaves as a dominant negative form of VHL, is more distant from both zebrafish genes. Accession numbers: NP_000542 (human); NP_033533 (mouse); NP_43468 (rat); XP_414447 (chicken); NP_001016367 (xenopus); CK360574 (zebrafish); ENSTRUP00000028576 (fugu); CAG06777 (tetraodon); ENSORLP00000001841 (medaka); EG536853 (minnow; Rutilus rutilus); CU063334 (rainbow trout; Oncorhynchus mykiss); EB687673 (dogfish; Squalus Acanthia); XM_001335821 (zebrafish Vhl-like); EG531650 (minnow Vhl-like); CU063690 (rainbow trout Vhl-like); (dogfish Vhl-like); NP_001004319 (human VLP).
- Figure S2. Synteny between chromosomal regions containing zebrafish and human VHL (JPG, 33 KB)
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Comparison of the chromosomal regions containing human VHL (chr 3p25.3, 8.6–10.6 Mb) and zebrafish vhl (chr 6, 27–29.4 Mb, Zv7) in Ensembl (www.ensembl.org) reveals synteny between both locations.
- Figure S3. vhl mutants display an increased number of L-plastin+ early macrophages (JPG, 59.2 KB)
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Immunohistochemical analysis of L-plastin protein levels reveals an increased number of L-plastin+ early macrophages in the vhl−∕− trunk at 7 dpf when compared to age-matched siblings. Original magnification 8 × 10.
- Video 1. vhl mutants display a hyperventilation response (WMV, 2.33 MB)
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Ventilation rate in siblings (upper panels) and vhl mutants (lower panels) at 9 dpf (lateral view, left panels) and 7 dpf (ventral view, right panels) in real time. See Document 1 for technical details.
- Video 2. TG(gata1:egfp) vhl mutants display an increased blood circulation (WMV, 2.29 MB)
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Blood circulation in the tail of a 6 dpf TG(gata1:egfp) sibling (upper panel) and vhl mutant (lower panel) in real time. Anterior is to the left. See Document 1 for technical details.
- Video 3. Sibling blood circulation at 9 dpf (bright field) (WMV, 955 KB)
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Bright field imaging of the blood circulation in the dorsal aorta at the level of the cloaca of a 9 dpf sibling. Ventral is up, anterior is to the left. See Document 1 for technical details.
- Video 4. vhl−∕− blood circulation at 9 dpf (bright field) (WMV, 1.79 MB)
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Bright field imaging of the blood circulation in the dorsal aorta at the level of the cloaca of a 9 dpf vhl mutant. Ventral is up, anterior is to the left. See Document 1 for technical details.
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