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.

Coarse and fine particulate matter (PM) were taken by a dichotomous sampler, and gas precursors were determined by a denuder sampler at two stations in central Taiwan. Water-soluble ionic constituents of PM and their precursor gases were analyzed by ionic chromatograph. In summer, the daytime/nighttime PM10 concentrations were 37 ± 10/41 ± 18 μg m(-3) and 36 ± 14/34 ± 18 μg m(-3) for Xitun and Jhushan, respectively. Average PM10 concentration in winter was 1.55 and 1.76 times that of summer for Xitun and Jhushan, respectively. PM mass concentrations were similar for both stations, although one station is located in the downtown area of Taichung, and the other is in a rural area with no heavy pollution sources. Water-soluble ionic species content was 38-53 % of PM2.5 and 43-48 % of PM10 mass concentration. HNO3, HCl, and SO2 were high in the daytime; the daytime-to-nighttime concentration ratio was 3.75-6.88 for HNO3,1.7-7.8 for HCl, and 1.45-2.77 for SO2. High NH3 levels were determined in the area, especially in winter, which could be a precursor of NH4 (+) to form particulate matter. In Xitun, motor vehicles downtown and in the industrial district could be sources of air pollution. In contrast, there are few industrial sources at Jhushan; therefore, the transport of air pollutants from upwind of other regions and the accumulation of pollutants could be important PM sources at Jhushan.