Characterization of the Zebrafish Homolog of Zipper Interacting Protein Kinase

Zipper-interacting protein kinase (ZIPK) is a conserved vertebrate-specific regulator of actomyosin contractility in smooth muscle and non-muscle cells. Murine ZIPK has undergone an unusual divergence in sequence and regulation compared to other ZIPK orthologs. In humans, subcellular localization is controlled by phosphorylation of threonines 299 and 300. In contrast, ZIPK subcellular localization in mouse and rat is controlled by interaction with PAR-4. We carried out a comparative biochemical characterization of the regulation of the zebrafish ortholog of ZIPK. Like the human orthologs zebrafish ZIPK undergoes nucleocytoplasmic-shuttling and is abundant in the cytoplasm, unlike the primarily nuclear rat ZIPK. Rat ZIPK, but not human or zebrafish ZIPK, interacts with zebrafish PAR-4. Mutation of the conserved residues required for activation of the mammalian orthologs abrogated activity of the zebrafish ZIPK. In contrast to the human ortholog, mutation of threonine 299 and 300 in the zebrafish ZIPK has no effect on the activity or subcellular localization. Thus, we found that zebrafish ZIPK functions in a manner most similar to the human ZIPK and quite distinct from murine orthologs, yet the regulation of subcellular localization is not conserved.     

5种新型杀菌剂对4种鱼的急性毒性及安全性评价

为评价5种新型杀菌剂的环境安全性,采用半静态法测定了其对4种鱼的急性毒性。结果表明,氟醚菌酰胺对斑马鱼、稀有鮈鲫和凤尾鲫的LC_(50)(96 h)均大于100 mg/L,对青鳉的LC_(50)(96 h)为35.25 mg/L,其急性毒性均为低毒;氟吡菌胺对斑马鱼、青鳉、稀有鮈鲫和凤尾鲫的LC_(50)(96 h)分别为1.49、0.56、0.35、1.04 mg/L,其对斑马鱼和凤尾鲫急性毒性为中毒,对青鳉和稀有鮈鲫为高毒;苯噻菌胺对斑马鱼、稀有鮈鲫和凤尾鲫的LC_(50)(96 h)均大于100 mg/L,对青锵的LC_(50)(96 h)为88.23mg/L,其急性毒性均为低毒;嘧菌酯对斑马鱼、青鳉、稀有鮈鲫和凤尾鲫的LC_(50)(96 h)分别为106.80、0.44、85.96、4.61 mg/L,其对斑马鱼和稀有鮈鲫急性毒性为低毒,对青鳉急性毒性为高毒,对凤尾鲫为中毒;醚菌酯对斑马鱼、青鳝、稀有鮈鲫和凤尾鲫的LC_(50)(96 h)分别为0.77、0.66、0.51、0.81 mg/L,其对4种鱼急性毒性均为高毒。明确这些药剂的环境安全性对指导其科学使用,保护环境生物均有较大意义。     

Genes and environment - striking the fine balance between sophisticated biomonitoring and true functional environmental genomics

This article provides an overview how the application of the gene profiling (mainly via microarray technology) can be used in different organisms to address issues of environmental importance. Only recently, environmental sciences, including ecotoxicology, and molecular biology have started to mutually fertilize each other. This conceptual blend has enabled the identification of the interaction between molecular events and whole animal and population responses. Likewise, striking the fine balance between biomonitoring and functional environmental genomics will allow legislative and administrative measures to be based on a more robust platform. The application of DNA microarrays to ecotoxicogenomics links ecotoxicological effects of exposure with expression profiles of several thousand genes. The gene expression profiles are altered during toxicity, as either a direct or indirect result of toxicant exposure and the comparison of numerous specific expression profiles facilitates the differentiation between intoxication and true responses to environmental stressors. Furthermore, the application of microarrays provides the means to identify complex pathways and strategies that an exposed organism applies in response to environmental stressors. This review will present evidence that the widespread phenomenon of hormesis has a genetic basis that goes beyond an adaptive response. Some more practical advantages emerge: the toxicological assessment of complex mixtures, such as effluents or sediments, as well as drugs seems feasible, especially when classical ecotoxicological tests have failed. The review of available information demonstrates the advantages of microarray application to environmental issues spanning from bacteria, over algae and spermatophytes, to invertebrates (nematode Caenorhabditis elegans, crustacea Daphnia spp., earthworms), and various fish species. Microarrays have also highlighted why populations of a given species respond differently to similar contaminations. Furthermore, this review points at inherent limits of microarrays which may not yet have been properly addressed, namely epigenetics, which may explain heritable variation observed in natural population that cannot be explained by differences in the DNA sequence. Finally, the review will address promising future molecular biological developments which may supersede the microarray technique.     

两种鱼类Hoxa-11基因片段克隆及5''端序列分析

为了分析脊椎动物从鳍到肢转变过程中基因的系统进化，采用PCR法克隆了斑马鱼和矛尾鱼的Hoxa-11基因片段，测序并进行序列分析。结果显示同源异型盒所在的外显子Ⅱ区和剪接位点是高度保守的，外显子I区又可分为四个亚区；中度保守区、可变区和两个高度保守区。从鱼到四足动物，Hoxa-11基因的主要变化是可变区的长度梯增且出现富含丙氨酸的区域。此外，在内含子中也发现了两个高度保守的DNA序列，其长度分别为35bp和16bp。     

Heterocyclic compounds as releasers of the fright reaction in the giant danioDanio malabaricus (Jerdon) (Cyprinidae,Ostariophysi, Pisces)

Fifty-nine pteridine, purine, and pyrimidine derivatives were tested with schools of the giant danioDanio malabaricus (Jerdon). The fright reaction was elicited by three pteridine derivatives: 2,6-diamino-4-oxodihydropteridine, isoxanthopterin, and 6-acetonyliso-xanthopterin. A minor effect could not be excluded for three purine derivatives: I-5-MP, IDP, and ITP.     

Fish‐Microarray: A Miniaturized Platform for Single‐Embryo High‐Throughput Screenings

Small molecule high‐throughput screenings are essential for the fields of drug discovery and toxicology. Such screenings performed on whole animals are more physiologically relevant leading to more predictive results. However, due to challenges in automation, high costs and absence of miniaturized solutions for animal‐based experiments, high throughput screenings based on animal models are still in its infancy. Here a platform for miniaturized high throughput whole‐organism screenings is presented. The new platform is based on patterns of hydrophilic spots separated by superhydrophobic borders. The difference in wettability of spots and borders generates the effect of discontinuous dewetting and allows for formation of arrays of microdroplets that incorporate single fish embryos. Due to the flat border‐less nature of the platform, the system is compatible with single‐step collection of embryos and pipetting‐free parallel addition of chemical libraries using the “sandwiching method.” The system is miniaturized and allows for incubation of embryos in volumes as low as 5 µL. Finally, the platform realizes surface tension based immobilization of single embryos inside of each microcompartment and permits high‐throughput microscopic analysis directly on the platform. Thus, this method combines the advantages of microarrays, such as high‐throughput and simplicity, with the power of in vivo experiments.     

Where and Why Modeling Amyotrophic Lateral Sclerosis

Over the years, researchers have leveraged a host of different in vivo models in order to dissect amyotrophic lateral sclerosis (ALS), a neurodegenerative/neuroinflammatory disease that is heterogeneous in its clinical presentation and is multigenic, multifactorial and non-cell autonomous. These models include both vertebrates and invertebrates such as yeast, worms, flies, zebrafish, mice, rats, guinea pigs, dogs and, more recently, non-human primates. Despite their obvious differences and peculiarities, only the concurrent and comparative analysis of these various systems will allow the untangling of the causes and mechanisms of ALS for finally obtaining new efficacious therapeutics. However, harnessing these powerful organisms poses numerous challenges. In this context, we present here an updated and comprehensive review of how eukaryotic unicellular and multicellular organisms that reproduce a few of the main clinical features of the disease have helped in ALS research to dissect the pathological pathways of the disease insurgence and progression. We describe common features as well as discrepancies among these models, highlighting new insights and emerging roles for experimental organisms in ALS.