PUBLICATION
A novel zebrafish jak2a(V581F) model shared features of human JAK2(V617F) polycythemia vera
- Authors
- Ma, A.C., Fan, A., Ward, A.C., Liongue, C., Lewis, R.S., Cheng, S.H., Chan, P.K., Yip, S.F., Liang, R., and Leung, A.Y.
- ID
- ZDB-PUB-090929-3
- Date
- 2009
- Source
- Experimental hematology 37(12): 1379-1386 (Journal)
- Registered Authors
- Leung, Anskar, Liongue, Clifford, Ward, Alister C.
- Keywords
- Zebrafish, hematopoiesis, polycythemia vera, Janus kinase 2
- MeSH Terms
-
- Amino Acid Sequence
- Amino Acid Substitution
- Animals
- Base Sequence
- Blotting, Western
- Disease Models, Animal
- Embryo, Nonmammalian/enzymology
- Embryo, Nonmammalian/metabolism
- Erythropoiesis/genetics
- Erythropoietin/genetics
- Erythropoietin/metabolism
- Gene Expression Regulation, Developmental
- Humans
- Janus Kinase 2/antagonists & inhibitors
- Janus Kinase 2/genetics*
- Janus Kinase 2/metabolism*
- Molecular Sequence Data
- Mutagenesis, Site-Directed
- Mutation*
- Polycythemia Vera/enzymology
- Polycythemia Vera/genetics
- Polycythemia Vera/pathology*
- Protein-Tyrosine Kinases/antagonists & inhibitors
- Protein-Tyrosine Kinases/genetics*
- Protein-Tyrosine Kinases/metabolism
- Pyrimidines/pharmacology
- Reverse Transcriptase Polymerase Chain Reaction
- STAT5 Transcription Factor/genetics
- STAT5 Transcription Factor/metabolism
- Sequence Homology, Amino Acid
- Sulfonamides/pharmacology
- Zebrafish/embryology
- Zebrafish/genetics
- Zebrafish/metabolism
- Zebrafish Proteins/antagonists & inhibitors
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism
- PubMed
- 19772888 Full text @ Exp. Hematol.
Citation
Ma, A.C., Fan, A., Ward, A.C., Liongue, C., Lewis, R.S., Cheng, S.H., Chan, P.K., Yip, S.F., Liang, R., and Leung, A.Y. (2009) A novel zebrafish jak2a(V581F) model shared features of human JAK2(V617F) polycythemia vera. Experimental hematology. 37(12):1379-1386.
Abstract
OBJECTIVE: The Janus kinase 2 (JAK2) is important for embryonic primitive hematopoiesis. A gain-of-function JAK2 (JAK2(V617F)) mutation in human is pathogenetically linked to polycythemia vera (PV). In this study, we generated a zebrafish orthologue of human JAK2(V617F) (referred herewith jak2a(V581F)) by site-directed mutagenesis and examined its relevance as a model of human PV. METHODS: Zebrafish embryos at 1-cell stage were injected with jak2a(V581F) mRNA (200pg/embryos). In some experiments, the embryos were treated with a specific JAK2 inhibitor TG101209. The effects of jak2a stimulation on hematopoiesis, jak/stat signaling and erythropoietin signaling were evaluated at 18hours-post-fertilization (hpf). RESULTS: Injection with jak2a(V581F) mRNA significantly increased erythropoiesis, as enumerated by flow cytometry based on gata1(+) population in dissociated Tg(gata1:gfp) embryos. The response was reduced by stat5.1 morpholino co-injection (Control: 4.37+/-0.08%; jak2a(V581F) injected: 5.71+/-0.07%, Co-injecting jak2a(V581F) and stat5.1 morpholino: 4.66+/-0.13%, p<0.01). jak2a(V581F) mRNA also up-regulated gata1 (1.83+/-0.08 fold, p=0.005), embryonic alpha (alphaeHb: 1.61+/-0.12 fold, p=0.049) and beta-hemoglobin gene expression (betaeHb: 1.65+/-0.13 fold, p=0.026) and increased stat5 phosphorylation. These responses were also ameliorated by stat5.1 morpholino co-injection or treatment with a specific JAK2 inhibitor TG101209. jak2a(V581F) mRNA significantly reduced erythropoietin gene [0.24+/-0.03 fold (p=0.006)] and protein expression (control: 0.633+/-0.11; jak2a(V581F) mRNA: 0.222+/-0.07 mIU/mL, p=0.019). CONCLUSION: The zebrafish jak2a(V581F) model shared many features with human PV and might provide us with mechanistic insights of this disease.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping