Name: zebrafish as a model to assess the teratogenic potential of nitrite
Uploaded: 2016-02-16T14:00:00.000+00:00
Duration: 7 min 29 s
Description: Watch this Scientific Journal Video about Zebrafish as a Model to Assess the Teratogenic Potential of Nitrite at JoVE.com

VK was funded by grants from the IUP Department of Biology and School of Graduate Studies and Research (Graduate Student Professional Development). CQD and TWS were supported by the IUP School of Graduate Studies and Research (Faculty Publication Costs/Incidental Research Expenses). We also thank members of the Diep laboratory for maintaining the zebrafish facility.

在本协议中所描述的程序进行的机构动物护理和使用委员会在宾夕法尼亚州印第安纳大学的批准。 1.收获胚胎<ol><li>在28.5℃，pH为7，500-1,500μS之间电导率维持斑马鱼，和14小时光照和10小时黑暗的24的光/暗周期。使用野生型菌株，如涂，AB或TU / AB混合。不同的菌株可能有不同的反应化学处理25。 </li><li>建立了鱼通过鱼加水系统到配合坦克交配收获鸡蛋的前一天晚上。添加雌雄鱼入罐和两个鱼分频器分开。每条鱼对将产生一系列的50-300蛋。为了确保有足够的蛋会产生，建立30双鱼。通常情况下，约50％在素配合年龄鱼对（6-9个月）将产生的蛋，导致每对多达200个鸡蛋，最大可达3的000蛋这个实验。 </li><li>第二天早晨，指示灯亮起后，取下分开始交配。检查水箱交配鸡蛋每15分钟。 </li><li>一旦鱼类产卵，用滤茶收获所有的胚胎，并将它们合并成E3缓冲液（5毫米氯化钠，0.17毫米氯化钾，0.33毫米氯化钙2，0.33毫米硫酸镁4）一个大容器。在1.5 HPF，删除和丢弃未受精的卵子在解剖显微镜下的塑料移液管。而受精卵是透明的34未受精的卵子是不透明的。 </li><li>将50胚胎成包含每个处理条件50毫升E3缓冲100×15毫米的玻璃培养皿。共有33菜肴需要11治疗条件和3个重复。 </li></ol> 2.治疗胚胎<ol><li>开展以2小时受精后（HPF）35治疗。孵育胚胎/幼虫在28.5℃，并在120高倍视野检查幼虫。在执行LEAST的统计分析各处理条件重复三次。 </li><li> 3培养皿（每片含50胚胎）在2 HPF，取出E3缓冲用移液管，并加入50毫升300毫米乙醇E3缓冲液稀释。覆盖的培养皿用Parafilm最小化的乙醇蒸发。 <ol><li>继续暴露胚胎乙醇22小时。然后取出用移液管乙醇。加入50mL的E3缓冲液中，摇动培养皿几次洗出乙醇。用移液管取出本届E3缓冲区，并重复洗涤步骤2次以上。 </li></ol></li><li> 3培养皿（每片含50胚胎）在2 HPF，取出E3缓冲用移液管，并加入50mL新E3缓冲。 </li><li> 9培养皿（每片含50胚胎）在2 HPF，取出E3缓冲用移液管，并加入50mL溶解在E3 1000缓冲mg / L的亚硝酸钠。事先确认使用的股票硝酸钠溶液浓度重氮化修饰（USEPA方法354.1）分光光度法28。 <ol><li>继续暴露3菜为46小时，三盘为70小时，三盘为94小时。每天更换和新鲜的亚硝酸盐溶液中的亚硝酸盐的解决方案。 </li><li>每次曝光时间后，取出用移液管中的亚硝酸盐。加入50mL的E3缓冲液中，摇动培养皿几次洗出亚硝酸盐。用移液管取出本届E3缓冲区，并重复洗涤步骤2次以上。 </li></ol></li><li> 3培养皿（每片含50胚胎）在2 HPF，取出E3缓冲用移液管，并加入50mL 200 mg / L的亚硝酸钠。 400，600，800重复此，与亚硝酸钠的1000毫克/升。 <ol><li>继续胚胎暴露于亚硝酸盐70小时。每天更换和新鲜的亚硝酸盐溶液中的亚硝酸盐的解决方案。 </li><li>曝光时间后，取出用移液管中的亚硝酸盐。加入50mL E3缓冲液，摇动几盘次，以洗出亚硝酸盐。用移液管取出本届E3缓冲区，并重复洗涤步骤2次以上。 </li></ol></li><li> 3培养皿（每片含50胚胎）在2 HPF，取出E3缓冲用移液管，并加入50mL溶解在E3 1000缓冲毫克/升硝酸钠。确认硝酸盐原液的浓度预先用镉还原（USEPA方法353.3）分光光度法28的变形例。 <ol><li>继续胚胎暴露于硝酸70小时。每天更换新鲜制成硝酸盐溶液的硝酸盐溶液。 </li><li>曝光时间后，取出用移液管硝酸盐。加入50mL的E3缓冲液中，摇动培养皿几次洗出硝酸盐。用移液管取出本届E3缓冲区，并重复洗涤步骤2次以上。 </li></ol></li><li>在曝光中的每一天，计数用立体显微镜在死胚胎/幼虫数。死亡的迹象包括缺乏心跳和血液循环，或观察1分钟后，缺乏活力的。除去死胚/幼虫用移液管以减小E3缓冲器的污染。 </li><li>当实验在120 HPF结束，安乐死的幼虫。 <ol><li>取出用移液管的E3缓冲区。然后加入50mL的0.2％的MS-222（缓冲到pH 7），等待10分钟。 </li><li>用移液管取出的MS-222。加入50mL的E3缓冲并乱舞的洗出的MS-222。 </li><li>用移液管取出E3缓冲器和加入20毫升4-％多聚甲醛（PFA）的固定幼虫。摇动的菜好几次。用移液管向幼虫转移到玻璃小瓶中有足够的PFA沿填充小瓶。存放在冰箱小瓶过夜（O / N）。 </li></ol></li><li>采取使用用数码相机一个立体镜固定幼虫的照片。使用30X的放大倍率，ISO 200和200毫秒的曝光时间。定向幼虫，使得前向乐英尺和背侧是对字段的顶部。 </li></ol>

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The authors have nothing to disclose.

这里介绍的方法证明了评估硝酸盐和亚硝酸盐的潜在致畸斑马鱼的效用。相比其他脊椎动物斑马鱼有优势，包括高繁殖力，体外受精，光学透明，快速发展。缺乏色素沉着（如斑马鱼的Casper 36）可用的突变体也有助于增强内脏器官的知名度。它也很容易产生与报告基因的转基因斑马鱼便于分析在活鱼37。因为斑马鱼基因组与人类保守的，从他们的研究中获得的信息可以导致在人中<...

畸变是由造成永久性的结构和功能异常，生长发育迟缓，或流产情况严重扰乱了1胚胎或胎儿的正常发育过程。它可以通过某些天然剂（致畸），其与胚胎发育以多种方式2干扰引起的。在人的胎儿发育，共同致畸如辐射，感染剂，有毒金属，和有机化工已报道引起epicanthic褶皱（弯曲手指或脚趾）通过形态发生错误的缺陷（皮肤褶皱中的上眼睑）和五指1。 理解致畸的分子机制是朝着显影治疗和预防的第一步。一些脊椎动物模型如非洲爪蛙（ 爪蟾 ）和斑马鱼（ 斑马鱼 ）已被用来确定受terat的分子途径ogens。以前的研究已经使用斑马鱼作为流行病学，毒理学和致畸3-7的典范。 Scholz等人 。考虑斑马鱼作为"金标准"环境毒性评估。这是由于在某种程度上，对斑马鱼胚胎，这允许它发生8研究者形象化发育缺陷的透明度。人类基因的大约70％的具有在斑马鱼直系同源物，使得斑马鱼的理想脊椎动物模型研究人类的缺陷9。 一些流行病学研究表明，硝酸盐和亚硝酸盐，通常存在于农场的食物和水，与出生缺陷或自然流产10,11相关联，而其它研究不支持此关联12。硝酸盐（NO 3 - ）和亚硝酸根（NO 2 - ）是在土壤和水自然存在。它们是氮植物的来源，并且是n的一部分itrogen周期13。食物，如绿豆，胡萝卜，南瓜，菠菜，甜菜，并从使用化肥高硝酸盐农场显著增强硝酸盐和亚硝酸盐7的水平。从高硝酸盐的食物和鱼高硝酸盐水（主要是从土壤中流失30）喂牛奶会导致人体消耗大量的硝酸盐和亚硝酸盐14。硝酸盐和亚硝酸盐也常用于食品保鲜，这大大增加了由人类12摄取的量使用。 硝酸盐和亚硝酸盐的最佳水平像血管稳态和功能，神经传递及免疫宿主防御机制13-15生理过程中发挥根本性的作用。然而，暴露于高水平的硝酸盐和亚硝酸盐的可能导致不利的影响，特别是在婴儿和儿童16。摄入硝酸盐是由微生物，并在第进一步转化为亚硝酸盐在口腔中由肠道菌群16 E胃肠道。 硝酸盐使婴儿在蓝色婴儿综合症高风险的血红蛋白氧化成高铁血红蛋白，其携氧能力18损害血红蛋白。这导致皮肤的蓝色延伸到在更严重的情况下外周组织。在其他症状组织结果抑制氧化，最严重导致昏迷和死亡19,20。类似症状中的婴儿和成人在较高浓度的硝酸21的观察。由于紫绀，头痛亚硝酸盐中毒导致高铁血红蛋白成人水平升高，呼吸障碍31，死如不及时治疗，由于涉及重要组织缺氧32,33并发症。 硝酸盐在较高水平的摄入也可以导致各种健康并发症。儿童糖尿病，反复腹泻和反复呼吸道感染患儿已与硝酸盐摄入11,17,22相连。长期暴露于高水平硝酸盐与排尿和脾出血有关。急性高剂量照射硝酸盐可导致像腹痛，肌肉无力，便血和尿，昏厥，甚至死亡11医疗条件范围广泛。在较高水平产前暴露于硝酸一直与神经管缺陷的肌肉骨骼相关的11。 最近的一份报告显示，治疗斑马鱼胚胎与亚导致卵黄囊水肿，颅面和轴向畸形和鱼鳔noninflation 5。在这项研究中，我们证明了治疗斑马鱼的胚胎与硝酸盐和亚硝酸盐，以确定他们的致畸潜力的方法。胚胎在不同浓度和不同的时间长度暴露于亚硝酸盐。乙醇用作阳性对照，因为它是一个既定的致畸23。 Ø乌尔法表明，两种高浓度，长的曝光时间为亚硝酸盐不利于生存和导致各种表型，从轻度（水肿），重度（毛发育缺陷）。因此，斑马鱼是直接在探索胚胎硝酸盐和亚硝酸盐的潜在致畸作用，以补充流行病学研究的有用模型。

<table><tbody><tr><td>DREL/2010 instrument</td><td>Hach</td><td>26700-03</td><td></td></tr><tr><td>Ethanol</td><td>Sigma-Aldrich</td><td>E7023</td><td></td></tr><tr><td>KIMAX glass Petri Dish</td><td>VWR</td><td>89001-244</td><td></td></tr><tr><td>MS-222</td><td>Sigma-Aldrich</td><td>E10521</td><td></td></tr><tr><td>NitraVer 5 Nitrate Reagent</td><td>Hach</td><td>14034-46</td><td></td></tr><tr><td>NitriVer 3 Nitrite Reagent</td><td>Hach</td><td>14065-99</td><td></td></tr><tr><td>Parafilm</td><td>Fisher Scientific</td><td>3-374-10</td><td></td></tr><tr><td>Paraformaldehyde</td><td>Sigma-Aldrich</td><td>158127</td><td></td></tr><tr><td>S6E stereomicroscope</td><td>Leica</td><td>10446294</td><td></td></tr><tr><td>Sodium nitrate</td><td>Fisher Scientific</td><td>S343</td><td></td></tr><tr><td>Sodium nitrite</td><td>Fisher Scientific</td><td>S347</td><td></td></tr><tr><td>Transfer pipets</td><td>Laboratory Products Sales</td><td>L320072</td><td></td></tr><tr><td>Glass vials</td><td>Fisher Scientific</td><td>03-338B</td><td></td></tr></tbody></table>

zebrafish as a model to assess the teratogenic potential of nitrite

在环境中的高硝酸盐水平可能导致在人体中的先天性缺陷或流产。据推测，这是由于硝酸根的由肠道和唾液细菌转化为亚硝酸盐。然而，在其他哺乳动物的研究中，高亚硝酸盐的含量不导致出生缺陷，尽管它们可能会导致差的生殖结局。因此，亚硝酸盐的致畸潜在不明确。这将是有脊椎动物模型系统轻松评估亚硝酸盐或任何其他感兴趣的化学物质的致畸作用非常有用。这里，我们证明斑马鱼（ 斑马鱼 ）的工具来筛选毒性和胚胎缺陷化合物。斑马鱼胚胎受精外，并有快速发展，使他们致畸研究的一个很好的模式。我们表明，增加曝光时间以负亚硝酸盐影响生存。增加亚硝酸盐的浓度也有不利影响的生存，而硝酸盐没有。对于胚胎塔ŧ生存亚硝酸盐暴露，可能会出现各种缺陷，包括心包和卵黄囊水肿，鱼鳔noninflation和颅面畸形。我们的结果表明，该斑马鱼是研究亚硝酸盐的致畸电位的方便的系统。这种方法可以容易地适用于测试其他化学品的早期脊椎动物发育的影响。

曝光至300mM乙醇22小时对存活率（数据未示出），与以前的报告5,23,26一致没有影响。这是意料之中的，因为乙醇是一种已知的致畸，并担任阳性对照。观测到的表型包括心包水肿，鱼鳔noninflation（图1），颅面畸形和发育迟缓（数据未示出）。 用亚硝酸盐处理导致轻度至对生存造成严重影响，这取决于曝光?...

Watch this Scientific Journal Video about Zebrafish as a Model to Assess the Teratogenic Potential of Nitrite at JoVE.com

Zebrafish as a Model to Assess the Teratogenic Potential of Nitrite

接触致畸可能导致出生缺陷。斑马鱼是确定的化学物质的致畸潜在有用的。我们通过暴露胚胎各级亚硝酸盐，并且也在曝光不同的时间证明斑马鱼的效用。我们表明，亚硝酸盐可能是有毒的，并导致严重的发育缺陷。

斑马鱼作为一种模型来评估亚硝酸盐的潜在致畸

High nitrate levels in the environment may&#160;result in congenital defects or miscarriages in humans. Presumably, this is due to the conversion of ...

zebrafish-as-a-model-to-assess-the-teratogenic-potential-of-nitrite

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Adverse Drug Effects and Chemical Toxicity

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9.4K 次观看.  Indiana University of Pennsylvania. 接触致畸可能导致出生缺陷。斑马鱼是确定的化学物质的致畸潜在有用的。我们通过暴露胚胎各级亚硝酸盐，并且也在曝光不同的时间证明斑马鱼的效用。我们表明，亚硝酸盐可能是有毒的，并导致严重的发育缺陷。 

视频: Zebrafish as a Model to Assess the Teratogenic Potential of Nitrite

Assessing Teratogenic Changes in a Zebrafish Model of Fetal Alcohol Exposure

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, including mental retardation due to disordered and damaged brain development. Fetal alcohol spectrum disorder (FASD) is a term used to cover a continuum of birth defects that occur due to maternal alcohol consumption, and occurs in approximately 4% of children born in the United States. With 50% of child-bearing age women reporting consumption of alcohol, and half of all pregnancies being unplanned, unintentional exposure is a continuing issue2. In order to best understand the damage produced by ethanol, plus produce a model with which to test potential interventions, we developed a model of developmental ethanol exposure using the zebrafish embryo. Zebrafish are ideal for this kind of teratogen study3-8. Each pair lays hundreds of eggs, which can then be collected without harming the adult fish. The zebrafish embryo is transparent and can be readily imaged with any number of stains. Analysis of these embryos after exposure to ethanol at different doses and times of duration and application shows that the gross developmental defects produced by ethanol are consistent with the human birth defect. Described here are the basic techniques used to study and manipulate the zebrafish FAS model.

In order to understand the molecular mechanisms of the ethanol-induced developmental damage, we have developed a zebrafish model of ethanol exposure and are exploring the physical, cellular, and genetic alterations that occur after ethanol exposure1. We then seek to find potential interventions and rapidly test them in this animal model.

在斑马鱼模型的胎儿酒精暴露评估致畸的变化

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, ...

科研

医学

Studying Neurobehavioral Effects of Environmental Pollutants on Zebrafish Larvae

Recent years more and more environmental pollutants have been proved neurotoxic, especially at the early development stages of organisms. Zebrafish larvae are a preeminent model for the neurobehavioral study of environmental pollutants. Here, a detailed experimental protocol is provided for the evaluation of the neurotoxicity of environmental pollutants using zebrafish larvae, including the collection of the embryos, the exposure process, neurobehavioral indicators, the test process, and data analysis. Also, the culture environment, exposure process, and experimental conditions are discussed to ensure the success of the assay. The protocol has been used in the development of psychopathic drugs, research on environmental neurotoxic pollutants, and can be optimized to make corresponding studies or be helpful for mechanistic studies. The protocol demonstrates a clear operation process for studying neurobehavioral effects on zebrafish larvae and can reveal the effects of various neurotoxic substances or pollutants.

A detailed experimental protocol is presented in this paper for the evaluation of neurobehavioral toxicity of environmental pollutants using a zebrafish larvae model, including the exposure process and tests for neurobehavioral indicators.

环境污染物对斑马鱼幼虫的神经行为影响研究

Recent years more and more environmental pollutants have been proved neurotoxic, especially at the early development stages of organisms. Zebrafish larvae ...

环境科学

Zebrafish Larvae as a Model to Evaluate Potential Radiosensitizers or Protectors

Zebrafish are extensively used in several kinds of research because they are one of the easily maintained vertebrate models and exhibit several features of a unique and convenient model system. As highly proliferative cells are more susceptible to radiation-induced DNA damage, zebrafish embryos are a front-line in vivo model in radiation research. In addition, this model projects the effect of radiation and different drugs within a short time, along with major biological events and associated responses. Several cancer studies have used zebrafish, and this protocol is based on the use of radiation modifiers in the context of radiotherapy and cancer. This method can be readily used to validate the effects of different drugs on irradiated and control (non-irradiated) embryos, thus identifying drugs as radio sensitizing or protective drugs. Although this methodology is used in most drug screening experiments, the details of the experiment and the toxicity assessment with the background of X-ray radiation exposure are limited or only briefly addressed, making it difficult to perform. This protocol addresses this issue and discusses the procedure and toxicity evaluation with a detailed illustration. The procedure describes a simple approach for using zebrafish embryos for radiation studies and radiation-based drug screening with much reliability and reproducibility.

The zebrafish has recently been exploited as a model to validate potential radiation modifiers. The present protocol describes the detailed steps to use zebrafish embryos for radiation-based screening experiments and some observational approaches to evaluate the effect of different treatments and radiation.

斑马鱼幼虫作为评估潜在放射增敏剂或保护剂的模型

Zebrafish are extensively used in several kinds of research because they are one of the easily maintained vertebrate models and exhibit several features ...

癌症研究

Rapid Evaluation of Toxicity of Chemical Compounds Using Zebrafish Embryos

The zebrafish is a widely used vertebrate model organism for the disease and phenotype-based drug discovery. The zebrafish generates many offspring, has transparent embryos and rapid external development. Zebrafish embryos can, therefore, also be used for the rapid evaluation of toxicity of the drugs that are precious and available in small quantities. In the present article, a method for the efficient screening of the toxicity of chemical compounds using 1-5-day post fertilization embryos is described. The embryos are monitored by stereomicroscope to investigate the phenotypic defects caused by the exposure to different concentrations of compounds. Half-maximal lethal concentrations (LC50) of the compounds are also determined. The present study required 3-6 mg of an inhibitor compound, and the whole experiment takes about 8-10 h to be completed by an individual in a laboratory having basic facilities. The current protocol is suitable for testing any compound to identify intolerable toxic or off-target effects of the compound in the early phase of drug discovery and to detect subtle toxic effects that may be missed in the cell culture or other animal models. The method reduces procedural delays and costs of drug development.

Zebrafish embryos are used for evaluating the toxicity of chemical compounds. They develop externally and are sensitive to chemicals, allowing detection of subtle phenotypic changes. The experiment only requires a small amount of compound, which is directly added to the plate containing embryos, making the testing system efficient and cost-effective.

利用斑马鱼胚胎对化合物毒性的快速评价

The zebrafish is a widely used vertebrate model organism for the disease and phenotype-based drug discovery. The zebrafish generates many offspring, has ...

A Manual Small Molecule Screen Approaching High-throughput Using Zebrafish Embryos

Zebrafish have become a widely used model organism to investigate the mechanisms that underlie developmental biology and to study human disease pathology due to their considerable degree of genetic conservation with humans. Chemical genetics entails testing the effect that small molecules have on a biological process and is becoming a popular translational research method to identify therapeutic compounds. Zebrafish are specifically appealing to use for chemical genetics because of their ability to produce large clutches of transparent embryos, which are externally fertilized. Furthermore, zebrafish embryos can be easily drug treated by the simple addition of a compound to the embryo media. Using whole-mount in situ hybridization (WISH), mRNA expression can be clearly visualized within zebrafish embryos. Together, using chemical genetics and WISH, the zebrafish becomes a potent whole organism context in which to determine the cellular and physiological effects of small molecules. Innovative advances have been made in technologies that utilize machine-based screening procedures, however for many labs such options are not accessible or remain cost-prohibitive. The protocol described here explains how to execute a manual high-throughput chemical genetic screen that requires basic resources and can be accomplished by a single individual or small team in an efficient period of time. Thus, this protocol provides a feasible strategy that can be implemented by research groups to perform chemical genetics in zebrafish, which can be useful for gaining fundamental insights into developmental processes, disease mechanisms, and to identify novel compounds and signaling pathways that have medically relevant applications.

The zebrafish is an excellent experimental organism to study vertebrate developmental processes and model human disease. Here, we describe a protocol on how to perform a manual high-throughput chemical screen in zebrafish embryos with a whole-mount in situ hybridization (WISH) read-out.

手册小分子屏幕接近高通量使用斑马鱼胚胎

Zebrafish have become a widely used model organism to investigate the mechanisms that underlie developmental biology and to study human disease pathology ...

生物学

Acute Kidney Injury Model Induced by Cisplatin in Adult Zebrafish

Cisplatin is commonly used as chemotherapy. Although it has positive effects in cancer-treated individuals, cisplatin can easily accumulate in the kidney due to its low molecular weight. Such accumulation causes the death of tubular cells and can induce the development of Acute Kidney Injury (AKI), which is characterized by a quick decrease in kidney function, tissue damage, and immune cells infiltration. If administered in specific doses cisplatin can be a useful tool as an AKI inducer in animal models. The zebrafish has appeared as an interesting model to study renal function, kidney regeneration, and injury, as renal structures conserve functional similarities with mammals. Adult zebrafish injected with cisplatin shows decreased survival, kidney cell death, and increased inflammation markers after 24 h post-injection (hpi). In this model, immune cells infiltration and cell death can be assessed by flow cytometry and TUNEL assay. This protocol describes the procedures to induce AKI in adult zebrafish by intraperitoneal cisplatin injection and subsequently demonstrates how to collect the renal tissue for flow cytometry processing and cell death TUNEL assay. These techniques will be useful to understand the effects of cisplatin as a nephrotoxic agent and will contribute to the expansion of AKI models in adult zebrafish. This model can also be used to study kidney regeneration, in the search for compounds that treat or prevent kidney damage and to study inflammation in AKI. Moreover, the methods used in this protocol will improve the characterization of tissue damage and inflammation, which are therapeutic targets in kidney-associated comorbidities.

This protocol describes the procedures to induce Acute Kidney Injury (AKI) in adult zebrafish using cisplatin as a nephrotoxic agent. We detailed the steps to evaluate the reproducibility of the technique and two techniques to analyze inflammation and cell death in the renal tissue, flow cytometry and TUNEL, respectively.

成人斑马鱼中西斯铂诱导的急性肾损伤模型

Cisplatin is commonly used as chemotherapy. Although it has positive effects in cancer-treated individuals, cisplatin can easily accumulate in the kidney ...

Nephrotoxin Microinjection in Zebrafish to Model Acute Kidney Injury

The kidneys are susceptible to harm from exposure to chemicals they filter from the bloodstream. This can lead to organ injury associated with a rapid decline in renal function and development of the clinical syndrome known as acute kidney injury (AKI). Pharmacological agents used to treat medical circumstances ranging from bacterial infection to cancer, when administered individually or in combination with other drugs, can initiate AKI. Zebrafish are a useful animal model to study the chemical effects on renal function in vivo, as they form an embryonic kidney comprised of nephron functional units that are conserved with higher vertebrates, including humans. Further, zebrafish can be utilized to perform genetic and chemical screens, which provide opportunities to elucidate the cellular and molecular facets of AKI and develop therapeutic strategies such as the identification of nephroprotective molecules. Here, we demonstrate how microinjection into the zebrafish embryo can be utilized as a paradigm for nephrotoxin studies.

Renal injuries incurred from nephrotoxins, which include drugs ranging from antibiotics to chemotherapeutics, can result in complex disorders whose pathogenesis remains incompletely understood. This protocol demonstrates how zebrafish can be used for disease modeling of these conditions, which can be applied to the identification of renoprotective measures.

肾毒素显微注射斑马鱼到模型急性肾损伤

The kidneys are susceptible to harm from exposure to chemicals they filter from the bloodstream. This can lead to organ injury associated with a rapid ...

行为的：评估成年斑马鱼神经毒性作用的测试电池

Neurotoxicity Assessment in Adult Danio rerio using a Battery of Behavioral Tests in a Single Tank

The presence of neuropathological effects proved to be, for many years, the main endpoint for assessing the neurotoxicity of a chemical substance. However, in the last 50 years, the effects of chemicals on the behavior of model species have been actively investigated. Progressively, behavioral endpoints were incorporated into neurotoxicological screening protocols, and these functional outcomes are now routinely used to identify and determine the potential neurotoxicity of chemicals. Behavioral assays in adult zebrafish provide a standardized and reliable means to study a wide range of behaviors, including anxiety, social interaction, learning, memory, and addiction. Behavioral assays in adult zebrafish typically involve placing the fish in an experimental arena and recording and analyzing their behavior using video tracking software. Fish can be exposed to various stimuli, and their behavior can be quantified using a variety of metrics. The novel tank test is one of the most accepted and widely used tests to study anxiety-like behavior in fish. The shoaling and social preference tests are useful in studying the social behavior of zebrafish. This assay is particularly interesting since the behavior of the entire shoal is studied. These assays have proven to be highly reproducible and sensitive to pharmacological and genetic manipulations, making them valuable tools for studying the neural circuits and molecular mechanisms underlying behavior. Additionally, these assays can be used in drug screening to identify compounds that may be potential modulators of behavior.
We will show in this work how to apply behavioral tools in fish neurotoxicology, analyzing the effect of methamphetamine, a recreational drug, and glyphosate, an environmental pollutant. The results demonstrate the significant contribution of behavioral assays in adult zebrafish to the understanding of the neurotoxicological effects of environmental pollutants and drugs, in addition to providing insights into the molecular mechanisms that may alter neuronal function.

作者聚焦：成年斑马鱼环境和药物诱导的行为变化评估 

Here, we present a comprehensive behavioral test battery, including the novel tank, Shoaling, and social preference tests, to effectively determine the potential neurotoxic effects of chemicals (e.g., methamphetamine and glyphosate) on adult zebrafish using a single tank. This method is relevant to neurotoxicity and environmental research.

斑马鱼作为一种模型来评估亚硝酸盐的潜在致畸

摘要

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此视频中的章节

在斑马鱼模型的胎儿酒精暴露评估致畸的变化

环境污染物对斑马鱼幼虫的神经行为影响研究

斑马鱼幼虫作为评估潜在放射增敏剂或保护剂的模型

利用斑马鱼胚胎对化合物毒性的快速评价

手册小分子屏幕接近高通量使用斑马鱼胚胎

成人斑马鱼中西斯铂诱导的急性肾损伤模型

肾毒素显微注射斑马鱼到模型急性肾损伤

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估