Zebrafish WNK lysine deficient protein kinase 1 (wnk1) affects angiogenesis associated with VEGF signaling

Ju-Geng Lai; Su-Mei Tsai; Hsiao-Chen Tu; Wen-Chuan Chen; Fong-Ji Kou; Jeng-Wei Lu; Horng-Dar Wang; Chou-Long Huang; Chiou-Hwa Yuh

doi:10.1371/journal.pone.0106129

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Zebrafish WNK lysine deficient protein kinase 1 (wnk1) affects angiogenesis associated with VEGF signaling

Journal article

Open access

Peer reviewed

Zebrafish WNK lysine deficient protein kinase 1 (wnk1) affects angiogenesis associated with VEGF signaling

Ju-Geng Lai, Su-Mei Tsai, Hsiao-Chen Tu, Wen-Chuan Chen, Fong-Ji Kou, Jeng-Wei Lu, Horng-Dar Wang, Chou-Long Huang and Chiou-Hwa Yuh

PloS one, Vol.9(8), pp.e106129-e106129

2014

DOI: 10.1371/journal.pone.0106129

PMCID: PMC4149531

PMID: 25171174

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Abstract

The WNK1 (WNK lysine deficient protein kinase 1) protein is a serine/threonine protein kinase with emerging roles in cancer. WNK1 causes hypertension and hyperkalemia when overexpressed and cardiovascular defects when ablated in mice. In this study, the role of Wnk1 in angiogenesis was explored using the zebrafish model. There are two zebrafish wnk1 isoforms, wnk1a and wnk1b, and both contain all the functional domains found in the human WNK1 protein. Both isoforms are expressed in the embryo at the initiation of angiogenesis and in the posterior cardinal vein (PCV), similar to fms-related tyrosine kinase 4 (flt4). Using morpholino antisense oligonucleotides against wnk1a and wnk1b, we observed that wnk1 morphants have defects in angiogenesis in the head and trunk, similar to flk1/vegfr2 morphants. Furthermore, both wnk1a and wnk1b mRNA can partially rescue the defects in vascular formation caused by flk1/vegfr2 knockdown. Mutation of the kinase domain or the Akt/PI3K phosphorylation site within wnk1 destroys this rescue capability. The rescue experiments provide evidence that wnk1 is a downstream target for Vegfr2 (vascular endothelial growth factor receptor-2) and Akt/PI3K signaling and thereby affects angiogenesis in zebrafish embryos. Furthermore, we found that knockdown of vascular endothelial growth factor receptor-2 (flk1/vegfr2) or vascular endothelial growth factor receptor-3 (flt4/vegfr3) results in a decrease in wnk1a expression, as assessed by in situ hybridization and q-RT-PCR analysis. Thus, the Vegf/Vegfr signaling pathway controls angiogenesis in zebrafish via Akt kinase-mediated phosphorylation and activation of Wnk1 as well as transcriptional regulation of wnk1 expression.

Phosphorylation

Zebrafish Proteins - metabolism

Vascular Endothelial Growth Factor Receptor-2 - metabolism

Zebrafish

Phosphatidylinositol 3-Kinases - metabolism

Vascular Endothelial Growth Factor A - metabolism

Vascular Endothelial Growth Factor Receptor-3 - metabolism

Neovascularization, Physiologic - physiology

Animals

Signal Transduction - physiology

Neovascularization, Pathologic - metabolism

Proto-Oncogene Proteins c-akt - metabolism

Protein-Serine-Threonine Kinases - metabolism

Details

Title: Subtitle: Zebrafish WNK lysine deficient protein kinase 1 (wnk1) affects angiogenesis associated with VEGF signaling
Creators: Ju-Geng Lai - Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan Town, Miaoli, Taiwan, ROC
Su-Mei Tsai - Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan Town, Miaoli, Taiwan, ROC
Hsiao-Chen Tu - Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan Town, Miaoli, Taiwan, ROC; Institute of Biotechnology, National Tsing Hua University, Hsinchu, Taiwan, ROC
Wen-Chuan Chen - Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan Town, Miaoli, Taiwan, ROC
Fong-Ji Kou - Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan Town, Miaoli, Taiwan, ROC
Jeng-Wei Lu - Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan Town, Miaoli, Taiwan, ROC
Horng-Dar Wang - Institute of Biotechnology, National Tsing Hua University, Hsinchu, Taiwan, ROC
Chou-Long Huang - Departments of Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America
Chiou-Hwa Yuh - Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan Town, Miaoli, Taiwan, ROC; College of Life Science and Institute of Bioinformatics and Structural Biology, National Tsing-Hua University, Hsinchu, Taiwan, ROC; Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan, ROC; College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC
Resource Type: Journal article
Publication Details: PloS one, Vol.9(8), pp.e106129-e106129
DOI: 10.1371/journal.pone.0106129
PMID: 25171174
PMCID: PMC4149531
NLM abbreviation: PLoS One
ISSN: 1932-6203
eISSN: 1932-6203
Publisher: Public Library of Science
Grant note: R01 DK059530 / NIDDK NIH HHS R01DK59530 / NIDDK NIH HHS
Language: English
Date published: 2014
Academic Unit: Nephrology; Internal Medicine
Record Identifier: 9984094311502771

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