整体原位杂交法研究斑马鱼(Danio rerio)胚胎发育过程中胸苷酸合成酶(TS)的表达

收集不同发育时期的斑马鱼胚胎,制备DIG标记的TS反义RNA探针,采用整体原位杂交方法研究胸苷酸合成酶(TS)基因在斑马鱼胚胎发育各期的时空表达状况.结果表明,在所取样的各个阶段,TS基因均有转录,但其部位不同,中囊胚过渡前后mRNA显现的部位仅存在于受精卵的动物极,10hpf时存在于整个胚胎的外围,至24hpf后集中...     

Neue Wege in die Ökotoxikologie: Quantitative Verhaltensmessungen an Fischen als Toxizitätsendpunkt

New Ways to Ecotoxicology: Quantitative Recording of Behaviour of Fish as Toxicity Endpoint The BehavioQuant system will be introduced which uses the integrative whole-organism response by automatized recording and quantifying the spontaneous behaviour of fish and other test organisms. While moving unrestricted within the test chamber, the movements of each single fish are recorded by video imaging, and their traces are digitized numerically by x-/y-coordinates. From the object traces several test parameters are calculated which give information about behavioural changes of the observed fishes. These various parameters are: the average swimming velocity (motility), the average swimming depth below surface, the inconstancy of motility, the frequency of turns, the standard frequency distributions of vertical and horizontal positions (can be used for quantification of a preference for dark or light habitats), and the distance behaviour. A continuously monitoring of up to 16 aquaria is possible. The statistic comparison between treated groups and control groups is done for each test parameter separately. The automatical evaluation of the continuous observation of surface waters results from a comparison of the actual behaviour data to the data of a reference period. Some examples for the applications of the BehavioQuant system will be demonstrated (effects of atrazine), and their results will be discussed. Behavioural changes of a single species can finally cause changes in biocenotic structures and functions. Therefore, it is necessary to include the results of quantitative behaviour measurements in ecotoxicological hazard assessment.     

Gene Expression Analysis for Exposure to Estrogenic Substances

Zebrafish (Danio rerio) were exposed with 17β‐estradiol (E2) and nonylphenol (NP) in different concentrations. Gene expression analysis was carried out by two different approaches. First, RT‐PCR experiments were performed for the examination of expression levels of the two marker genes vitellogenin and aromatase. This approach showed a significant increase in the expression of the vitellogenin gene in exposed male fish (500 ng/L 17β‐estradiol and 250 μg/L nonylphenol). This egg yolk protein is usually not synthesized in male vertebrates. A slight decrease of expression of the aromatase gene was observed in exposed female zebrafish. Aromatase is known to catalyze the conversion of androgens to estrogens. Second, DNA microarray experiments were carried out, which allow the simultaneous examination of the expression levels of a great number of marker genes. The microarray experiments resulted in an up‐regulation of vitellogenin up to 850‐fold. In addition, several other genes were identified to be up‐regulated by estrogens, for example the high mobility group box protein ssrp1 (78‐fold) or the chaperonin containing t‐complex polypeptide 1, beta subunit cctb (22‐fold).     

Sonophotocatalysis of endocrine-disrupting chemicals

Sonolysis and photolysis often exhibit synergistic effects in the degradation of organic molecules. An assay of fish oocyte maturation provides an appropriate experimental system to investigate the hormonal activities of chemical agents. Oocyte maturation in fish is triggered by maturation-inducing hormone (MIH), which acts on receptors on the oocyte surface. A synthetic estrogen, diethylstilbestrol (DES), possesses inducing activity of fish oocyte maturation, and a widely used biocide, pentachlorophenol (PCP), exhibits a potent inhibitory effect on fish oocyte maturation. In this study, the effects of the combined treatment by sonolysis with photolysis (sonophotocatalysis) to diminish the hormonal activity of DES and the maturation preventing activity of PCP was examined. By sonophotocatalysis, hormonal activity of DES was completely lost within 30min and the inhibiting activity of PCP was lost within 120min. These results demonstrated that sonophotocatalysis is effective for diminishing the endocrine-disrupting activity of chemical agents.     

热休克诱导斑马鱼异源三倍体的研究

于１９９７年５月在广东省珠海市集斑马鱼和豹纹斑马鱼,采用热休克方法研究阻止第三极体排放诱导异源三倍体斑马鱼的适宜条件。按正交实验方案组合诱导参数,结果表明：在卵受精后２ｍｉｎ,采用３９℃持续处理２ｍｉｎ,三倍化率可达５３．８％,原肠期相对存活率为９１．０％,尾芽期相对存活率为９１％,孵化期相对存活率为９１．１％。本研究分析了热休克处理的参数与三倍体出现率和胚胎相对存活率的关系,并得出诱导参数中起始     

The impact of musk ketone on reproduction in zebrafish (Danio rerio)

Nitro musks are a group of nitrated benzenes mainly used by the fragrance industry as a substitute for natural musk. Two of the most common nitro musks, musk xylene and musk ketone, have been detected in water, fish, human adipose tissue, as well as in human breast milk. In this study, female zebrafish were dietary-exposed to musk ketone, prior to spawning with unexposed males. The fish were exposed for 8 weeks at two different dose levels. Accumulation of the compound in the fish as well as reproductive success were studied. Exposed females had reduced body weight and length, as well as reduced liver- and gonad somatic index. The results from the reproduction study showed a dose-dependent reduction in fecundity. Early life-stage mortality was increased and the median survival time was reduced. In addition, an embryo/larvae toxicity test was performed, using newly fertilized zebrafish eggs from unexposed parental fish. Eggs were exposed to a series of different concentrations of musk ketone via the surrounding water. A NOEC value of 10 μg/l and a LOEC value of 33 μg/l was determined. Conclusively, the present study clearly shows that musk ketone negatively affects reproduction and early life-stage survival in zebrafish.     

Polystyrene nanoplastics aggravated ecotoxicological effects of polychlorinated biphenyls in on zebrafish (Danio rerio) embryos

In view of the accumulation of nanoplastics (NPs) in the food chain of environment and animals, and the good adsorption properties of nano-plastics to toxic substances, it is necessary to explore the influence of NPs in living organisms. In this study, single and joint toxicological effects of polystyrene nanoplastics (PS-NPs, size 80 nm) and polychlorinated biphenyls (PCBs), were explored in freshwater aquatic animal model zebrafish (Danio rerio). Our study found that exposure to single PS-NPs induced mild acute toxicity, albeit the combined exposure of PS-NPs and polychlorinated biphenyls aggravated the toxicity of PCBs in a dose-dependent manner. Results from gene expression profiling showed that NPs exposure could activate detoxification process, resulting in a slight up-regulation of antioxidant genes (sod1, gstp1), bone development genes (bmp2, bmp4) and cardiac gene (tbx20); while PCBs suppressed the detoxification through down-regulation of these genes, and the addition of NPs will exacerbate the impact of PCBs on gene suppression. Importantly, the results of in vivo purification experiments found that NPs showed prolonged retention in liver, intestine and gills of zebrafish and they might have crossed biological barrier and accumulate in lipid-rich tissues and excretion does not appear as the significant pathway for their elimination. In conclusion, the toxic effects of polychlorinated biphenyls on chorionic protected embryos were not significant as zebrafish chorion plays an important role in resisting the invasion of pollutants; PCBs can seriously damage the bone and heart development of zebrafish, while the presence of NPs significantly enhanced the toxicity of PCBs in zebrafish, which is an alarming concern for growing NPs levels and ecological safety in aquatic environment.