Name: gavaging adult zebrafish
Uploaded: 2013-08-11T14:00:00
Description: Watch this Scientific Journal Video about Gavaging Adult Zebrafish at JoVE.com

洛克菲勒大学生物科学中心比较为这个项目提供了支持。我们感谢詹妮尔Monnas的技术援助。

1。准备麻醉解决方案<ol><li>制备溶液150 mg / L的MS-222系统中的水，无论是从储备溶液或粉末形式。 </li><li>验证的pH值，用pH计。 </li><li>用碳酸氢钠缓冲液根据需要，直到pH值在7-7.5之间。 </li><li>准备从MS-222为鱼恢复英寸恢复时间通常小于1分钟的系统水的罐。 </li></ol> 2。准备灌胃装置<ol><li>切清晰的22-G植入管从口腔到肠道灯泡的鱼的长度大约多一点。最初，我们建议每个新组正...

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B., Chapman, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Preparing Your Own Fish Feeds. , (2009).</li><li>Gomez-Gil, B., Cabanillas-Ramos, J., Paez-Brambila, A., Roque, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standardization+of+the+bioencapsulation+of+enrofloxacin+and+oxytetracycline+in+Artemia+fransciscana+Kellogg.">Standardization of the bioencapsulation of enrofloxacin and oxytetracycline in Artemia fransciscana Kellogg.</a> Aquaculture. 196, 1-12 (1906).</li><li>Langdon, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microparticle+types+for+delivering+nutrients+to+marine+fish+larvae.">Microparticle types for delivering nutrients to marine fish larvae.</a> Aquaculture. 227, 259-275 (2003).</li><li>Langdon, C., Nordgreen, A., Hawkyard, M., Hamre, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+wax+spray+beads+for+delivery+of+low-molecular+weight,+water+soluble+nutrients+and+antibiotics+to+Artemia.">Evaluation of wax spray beads for delivery of low-molecular weight, water soluble nutrients and antibiotics to Artemia.</a> Aquaculture. 284, 151-158 (2008).</li><li>Zang, L., Morikane, D., Shimada, Y., Tanaka, T., Nishimura, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Novel+Protocol+for+the+Oral+Administration+of+Test+Chemicals+to+Adult+Zebrafish.">A Novel Protocol for the Oral Administration of Test Chemicals to Adult Zebrafish.</a> Zebrafish. 8, 203-210 (2011).</li><li>Goldsmith, J. 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意义这种技术是一种改进，并提供了几个优点，在先前所描述的技术用于gavaging成年斑马鱼。首先，利用海绵抑制鱼，一个人可以执行的程序，而不是先前所描述的技术，需要两个人。7二，比静脉导管，导管植入管更灵活，它可以帮助减少二次创伤所致穿孔同样使用钝头的注射器。7,8三，通过使用透明管，该解决方案可以轻松地进行可视化进入鱼。最后，灌胃?...

目前管理化合物口服斑马鱼的方法包括管理化合物涂在饲料，混合成明胶的饮食1，盐水虾2 bioencapsulated，混合脂质的封装饮食3，喷蜡珠4，并通过无麸质饮食。限制这些方法包括口服高浸出率和个别鱼不完整或不可预知的化合物消费。这些变量是有问题的，因为在传染性疾病研究，知道鱼的感染剂量可能是学习成功的关键。另外，以前的工作已表明，某些化合物给药水浴引起中毒性病变肠道的影响，可能是所研究的发生之前，斑马鱼鳃。 灌胃是一个标准的方法，用于其他实验室动物物种的精确管理的产品数量AKnown集中研究在生物医学和制药研究。直到最近，在文献中被描述为gavaging斑马鱼的方法。描述的技术之一是两个人的方法，使用24-G连接至2-20微升的移液管，以提供溶液5μl的导管鞘7中所描述的方法的死亡率是8.7％和39％，在第一和第二试验，分别大部分死亡率归因于妊娠女性。第二灌胃描述的技术使用钝头灌胃注射器辖5微升解决medakas的青鳉 。8对死亡率的信息并没有提供，进行灌胃，没有描述的确切过程。根据动物护理和实验室斑马鱼，鱼的身体重量的1％到加拿大会可以通过灌胃给药9,10，我们的目标是开发一个可重复的，安全，高效的方法提供精确的体积COM磅的成年斑马鱼的男女双方口头和再生产的各个阶段。此过程将是适用于任何研究中，需要准确的化合物口服给药。

<table border="1">
<tbody>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Equipment</td>
 </tr>
 <tr>
 <td>A sponge</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>1-cc luer lock syringe</td>
 <td>Becton Dickinson &amp; Co</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>22-G Micro-Renathane Implantation tubing (cut)</td>
 <td>Braintree Scientific, Inc.</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>1 22-G needle (cut, if needed)</td>
 <td>Becton Dickinson &amp; Co</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>pH meter</td>
 <td>Hanna probe</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>1 scalpel blade</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>[header]</td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Reagent/Material</td>
 </tr>
 <tr>
 <td>MS-222 (Finquel)</td>
 <td>Argent Laboratories</td>
 <td></td>
 <td></td>
 </tr>
 </tbody>
</table>

gavaging adult zebrafish

斑马鱼已成为一个重要的在体内模型在生物医学研究。有效方法，必须开发和利用提供解决方案，为科研的化合物或试剂。目前管理化合物口服成年斑马鱼的方法是不准确由于变异自愿消费的鱼。一个灌胃的程序开发传染性病原体对斑马鱼的研究在生物医学研究提供精确的数量。超过6个月的年龄的成年斑马鱼麻醉与150 mg / L的缓冲MS-222和，灌胃5微升的解决方案，采用柔性导管植入管连接到一个切割22-G针尖。下降到斑马鱼的口腔直到管子的前端延伸过去鳃（约1厘米）的软管。然后将溶液缓慢注入到肠道内。此方法是有效的88％的时间里，鱼没有事故恢复。这程序也是有效的，作为一个人就可以在一小时内灌胃给药20-30鱼。此方法可以用于精确地管理代理传染病的研究，或成年斑马鱼中的其它化合物的研究。

这两种鱼类，包括妊娠女性两性成功灌胃（ 图1）。一个成功的过程不到一分钟，没有看到退出鳃或口的液体。油管进入容易，没有力量，没有看到血后拔除尿管。这个程序是快速的，需要约10分钟，灌胃给药3〜4的鱼，鱼每30-45秒的平均。 斑马鱼不应该被灌胃超过5μl的任何解决方案。管饲法用10微升的解决方案导致只有50％的成功率（ 图2）。灌胃10?...

Watch this Scientific Journal Video about Gavaging Adult Zebrafish at JoVE.com

Gavaging Adult Zebrafish

Gavaging成年斑马鱼

越来越多地使用斑马鱼作为动物模型的发展需要有效的方法交付已知化合物和解决方案的数量。灌胃下述过程可用于口服给药的精确体积的溶液可靠，安全和有效的成年斑马鱼。

The zebrafish has become an important in vivo model in biomedical research. Effective methods must be developed and utilized to deliver compounds or ...

Research

JoVE Journal

Biology

Transplantation of Whole Kidney Marrow in Adult Zebrafish

在成年斑马鱼的全肾骨髓移植

Hematopoietic stem cells (HSC) are a rare population of pluripotent cells that maintain all the differentiated blood lineages throughout the life of an ...

科研

生物学

Retro-orbital Injection in Adult Zebrafish

在成年斑马鱼的眼窝射出

Drug treatment of whole animals is an essential tool in any model system for pharmacological and chemical genetic studies. Intravenous (IV) injection is ...

Dissection of Organs from the Adult Zebrafish

剖析器官的成年斑马鱼

Over the last 20 years, the zebrafish has become a powerful model organism for understanding vertebrate development and disease.  Although experimental ...

Intraperitoneal Injection into Adult Zebrafish

腹腔注射到成年斑马鱼

A convenient method for chemically treating zebrafish is to introduce the reagent into the tank water, where it will be taken up by the fish. However, ...

Dissection of the Adult Zebrafish Kidney

成年斑马鱼肾脏的解剖

Researchers working in the burgeoning field of adult stem cell biology seek to understand the signals that regulate the behavior and function of stem ...

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

幼虫，少年和成年斑马鱼的心脏解剖，斑马鱼</em

Zebrafish have become a beneficial and practical model organism for the study of embryonic heart development (see recent reviews1-6), however, work ...

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

在体内电穿孔Morpholinos到成年斑马鱼视网膜

Many devastating inherited eye diseases result in progressive and irreversible blindness because humans cannot regenerate dying or diseased retinal ...

In vivo Electroporation of Morpholinos into the Regenerating Adult Zebrafish Tail Fin

In vivo Electroporation of Morpholinos into the Regenerating Adult Zebrafish Tail Fin

再生的成年斑马鱼尾鳍在吗啉代入体内电穿孔

Certain species of urodeles and teleost fish can regenerate their tissues. Zebrafish have become a widely used model to study the spontaneous regeneration ...

Blood Collection for Biochemical Analysis in Adult Zebrafish

在成年斑马鱼的生化分析采血车

The zebrafish has been used as an animal model for studies of several human diseases. It can serve as a powerful preclinical platform for studies of ...