斑马鱼GSTM单核苷酸多态性与低温耐受性的相关分析

为分析谷胱甘肽硫-转移酶M基因(glutathione S-transferases M,GSTM)与鱼类低温耐受性的相关性,本实验运用PCR-SSCP技术研究了130尾斑马鱼(Danio rerio)GSTM基因5?UTR、3?UTR和第一内含子序列的单核苷酸多态性(Single nucleotide polymorphisms,SNPs),同时分析了筛选到的基因型与其低温耐受性状的关联性。结果显示,在其5UTR区域检测到AB、BC、AC 3种基因型个体,共A、B、C 3个等位基因,其观测杂合度和期望杂合度分别是1.000和0.570,多态信息含量为0.472,所检测群体在该座位偏离了Hardy-Weinberg平衡;第1内含子中检测到DD、DE和EE 3种基因型个体,共D、E两个等位基因,其观测杂合度和期望杂合度分别为0.408和0.477,多态信息含量为0.362,群体在该座位符合Hardy-Weinberg平衡。3'UTR区域中没有发现多态性。上述2个SNP座位与斑马鱼低温耐受性状的关联分析结果表明,5'UTR区3种基因型与低温耐受性状均没有显著相关性(χ2=4.029,P0.05)。而第1内含子3种基因型与低温耐受性状显著相关(χ2=8.498,P0.05):DD基因型在耐低温群体中占优势(50.00%),并表现为对受低温胁迫斑马鱼的保护性因素(OR=0.520,95%CI=0.255–1.061),而DE基因型在不耐低温群体中占优势(51.31%),表现为低温胁迫下斑马鱼的危险因素(OR=3.012,95%CI=1.413–6.419)。研究结果为GSTM基因SNPs位点与斑马鱼低温耐受性能关联分析提供了依据,也将为海水经济鱼类抗寒标记筛选育种提供参考。     

3-甲基胆蒽对斑马鱼组织病理效应的初步研究

分别用25、50、100、200和400μg/L的3-甲基胆蒽(3-MC)对斑马鱼进行水浴染毒,于第7天和第14天采集各染毒组和对照组活鱼的肝、鳃、心、肠和肾,进行病理组织学检查。结果发现,一定剂量的3-MC能导致肝细胞变性,胞浆内出现脂肪滴,严重时肝细胞坏死解体,界限模糊;心肌细胞萎缩坏死;肠粘膜上皮细胞纹状缘不整齐,有的脱落;鳃丝结构异常,鳃小片表面粗糙,上皮细胞不完整;肾小管上皮细胞肿胀,细胞界限不清,管腔内均质红染。供试斑马鱼的病理组织变化与3-MC的作用剂量和作用时间存在一定的关系。     

Effects of base analogues 5-bromouracil and 6-aminopurine on development of zebrafish Danio Rerio

Zebrafish （Danio rerio） genetic screens allow isolation of a wide array of problems in vertebrate biology. The effects of base analogues 5-bromouracil and 6-aminopurine on the development of zebrafish embryos are reported for the first time in this study. The early development of the zebrafish embryos was little affected by 5-bromouracil and 6-aminopurine, while the late development （organogenesis） was significantly impaired. Embryos exposed to 5-bromouracil mainly showed curled tail, wavy body, golden pigmentation and the mouth with protruding lower jaw. 6-aminopurine-treated embryos had defective anterior structures, curled tails and wavy body. RAPD analysis showed that the majority of 5-bromouracil-and 6-aminopurine-treated larvae and fish shared banding patterns in common with the control, suggesting that most mutagenesis induced by these agents are point mutations. However, some fish derived from 5-bromouracil-treated embryos had golden（gol） pigmentation; and RAPD analysis revealed that their band patterns differed from those of the control.Possibly, 5-bromouracil can occasionally cause relatively extensive changes in the fish genome. The results of this study may provide valuable help for detailed studies of mutagenesis.     

苦地胆内酯对嗜水气单胞菌感染斑马鱼保护效果的研究

在饲料中添加不同比例的苦地胆内酯,饲喂斑马鱼成鱼14 d后,采用创伤感染法构建斑马鱼嗜水气单胞菌感染模型,研究苦地胆内酯对嗜水气单胞菌感染斑马鱼的保护效果.结果显示:饲喂含苦地胆内酯0.4%、0.8%、1.6%饲料的斑马鱼,其存活率显著高于阳性对照组(p值<0.05);0.8%和1.6%的苦地胆内酯对斑马鱼的保护率分别为(73.91±6.52)%和(73.91±11.30)%,与0.1%恩诺沙星保护率(76.08±7.53)%相比无显著差异(p值>0.05).一定剂量的苦地胆内酯对嗜水气单胞菌感染斑马鱼具有保护效果.     

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...     

三氯生对雌性斑马鱼肝胰脏凋亡相关基因表达的影响

【目的】研究三氯生(TCS)对雌性硬骨鱼类肝胰脏损伤的相关分子机制。【方法】采用半静态水体接触染毒法,将雌性斑马鱼(Danio rerio)暴露于0、0.017、0.034、0.068 mg/L的TCS溶液42 d,采用普通PCR和实时荧光定量PCR技术测定雌性斑马鱼肝胰脏凋亡相关基因(Bcl-2、p53、MDM2、Bax)的表达情况。【结果】0.017~0.068 mg/L组的雌性斑马鱼肝胰脏Bcl-2、MDM2和Bax基因的表达极显著下调(P<0.01),p53基因的表达极显著上调(P<0.01)。【结论】0.017~0.068 mg/L的TCS显著影响雌性斑马鱼肝胰脏凋亡相关基因的表达,促进雌性斑马鱼肝胰脏细胞凋亡的发生。     

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.     

miR-430对斑马鱼原始生殖细胞迁移和性腺发育影响的初步研究

为了探究microRNAs(miRNAs)在斑马鱼生殖细胞发育过程中所起的作用,本研究在斑马鱼胚胎期过表达miR-430,结果发现原始生殖细胞(Primordial germ cells,PGCs)迁移出现紊乱,并且通过qRT-PCR发现nanos3、piwil2及tdrd7等生殖质基因的表达水平显著下调.此外,在斑马...     

Effects of base analogues 5-bromouracil and 6-aminopurine on development of zebrafish Danio Rerio

Zebrafish (Danio rerio) genetic screens allow isolation of a wide array of problems in vertebrate biology. The effects of base analogues 5-bromouracil and 6-aminopurine on the development of zebrafish embryos are reported for the first time in this study. The early development of the zebrafish embryos was little affected by 5-bromouracil and 6-aminopurine, while the late development (organogenesis) was significantly impaired. Embryos exposed to 5-bromouracil mainly showed curled tail, wavy body, golden pigmentation and the mouth with protruding lower jaw. 6-aminopurine-treated embryos had defective anterior structures, curled tails and wavy body. RAPD analysis showed that the majority of 5-bromouracil- and 6-aminopurine-treated larvae and fish shared banding patterns in common with the control, suggesting that most mutagenesis induced by these agents are point mutations. However, some fish derived from 5-bromouracil-treated embryos had golden (gol) pigmentation; and RAPD analysis revealed that their band patterns differed from those of the control. Possibly, 5-bromouracil can occasionally cause relatively extensive changes in the fish genome. The results of this study may provide valuable help for detailed studies of mutagenesis. This work was supported by Ministry of Science and Technology (MOST) of China (G 1999012005).     

斑马鱼体细胞核移植-头肾细胞和尾鳍细胞核移入成熟具核卵子发育能力的初步研究

将经直接消化的斑马鱼头肾细胞或经短暂培养的尾鳍细胞核移植到同种成熟具核卵子中 ,移核卵可发育到原肠晚期 ;将移核卵发育成的囊胚细胞核作供体进行第 2次核移植 ,受体卵可发育到尾芽期或肌肉感应期。流式细胞仪测定 DNA含量表明 ,受体卵发育形成的囊胚细胞 DNA含量与正常囊胚细胞 DNA含量相同 ,由此可知 ,移核卵中的雌原核没有参与移核胚胎的发育。