ABSTRACT
This study was performed to determine the ecosystem health of an urban stream using DNA damage biomarkers of sentinel fish species along with chemical analyses of water and sediments in the impacted and reference zones. The results based on single-cell gel electrophoresis showed that the extent of DNA damage was significantly elevated in the polluted zone. The severity of DNA damage in the impacted zone was largely attributed to high levels of heavy metals, such as As, Zn, Pb, and Ni. The DNA damage biomarker in benthic fish was sensitive to habitat change, which suggested that this fish species and biomarker are excellent tools for risk assessment. The acute toxicity tests demonstrated that SCGE was sufficient to reflect the different genotypes and pollution statuses and to propose the possibility of zebrafish erythrocytes as competitive materials.
Subject(s)
Cyprinidae/genetics , DNA Damage , Ecosystem , Environmental Monitoring/methods , Rivers/chemistry , Water Pollutants, Chemical/toxicity , Zebrafish/genetics , Animals , Biomarkers/metabolism , Comet Assay , Ecotoxicology , Geologic Sediments/chemistry , Metals, Heavy/analysis , Metals, Heavy/toxicity , Mutagenicity Tests , Risk Assessment , Water/chemistry , Water Pollutants, Chemical/analysisABSTRACT
Siah2 is a zebrafish homologue of mammalian Siah family. Siah acts as an E3 ubiquitin ligase that binds proteins destined for degradation. Extensive homology between siah and Drosophila Siah homologue (sina) suggests their important physiological roles during embryonic development. However, detailed functional studies of Siah in vertebrate development have not been carried out. Here we report that Siah2 specifically augments nodal related gene expression in marginal blastomeres at late blastula through early gastrula stages of zebrafish embryos. Siah2 dependent Nodal signaling augmentation is confirmed by cell-based reporter gene assays using 293T cells and 3TPluciferase reporter plasmid. We also established a molecular hierarchy of Siah as a upstream regulator of FoxH1/Fast1 transcriptional factor in Nodal signaling. Elevated expression of nodal related genes by overexpression of Siah2 was enough to override the inhibitory effects of atv and lft2 on the Nodal signaling. In particular, E3 ubiquitin ligase activity of Siah2 is critical to limit the duration and/or magnitude of Nodal signaling. Additionally, since the embryos injected with Siah morpholinos mimicked the atv overexpression phenotype at least in part, our data support a model in which Siah is involved in mesendoderm patterning via modulating Nodal signaling.
Subject(s)
Nuclear Proteins/physiology , Signal Transduction , Ubiquitin-Protein Ligases/physiology , Zebrafish/embryology , Animals , Body Patterning , Embryo, Nonmammalian/enzymology , Embryonic Development , Forkhead Transcription Factors/metabolism , Gene Expression Regulation, Developmental , HEK293 Cells , Humans , Left-Right Determination Factors/physiology , Zebrafish Proteins/metabolism , Zebrafish Proteins/physiology , beta Catenin/metabolismABSTRACT
In zebrafish embryos, bilateral symmetry is broken by asymmetric nodal flow generated in Kupffer's vesicle (KV), the transient cilia-rich organ, analogous to the mouse node. Asymmetric nodal flow induces the asymmetric expression of several genes, which are critical for the determination of correct LR body patterning. seson encoding three consecutive C2H2 zinc finger protein is predominantly expressed in the cilia-rich organs including KV. Inhibition of its function by the injection of a seson-specific MO inhibited the left-side biased expression of spaw, and resulted in randomization of the heart, gut looping and brain laterality. Disruption of the LR patterning in seson morphants appeared to be due to severe cilia defects in KV. Seson function was also required for ciliogenesis in other tissues such as the pronephros and olfactory organs. Collectively, our data suggest that Seson has critical roles in ciliogenesis and LR body axis patterning.
