modified intraperitoneal injection of doxorubicin into adult zebrafish

优化腹腔注射用于成年斑马鱼化合物递送

Zebrafish (Danio rerio) has gained attention as an experimental model for studying human diseases because this animal encompasses high gene and organ homology to humans, external&#160;fertilization, ease of genetic manipulation, and body transparency into early maturity, which facilitates a myriad of imaging applications1. Unlike the straightforward process of delivering drugs directly to the water for zebrafish embryos and larvae, administering drugs to adult zebrafish presents a more intricate and challenging endeavor2.
In adult fish, compounds can be delivered through passive drug delivery techniques, such as direct administration into the water, or through oral drug delivery methods like gavaging2. Other approaches include coating fish food with the compounds and subsequently feeding the fish3, and direct administration of water-insoluble medications at a predetermined concentration, including retro-orbital or intraperitoneal injections4,5. Intraperitoneal administration is preferred for in vivo studies of disease models due to its distinct pharmacokinetic advantages6. This method provides a high drug concentration and an extended half-life within the peritoneal cavity, offering an effective route for drug delivery7,8. The approach is commonly utilized in research settings to ensure optimal drug absorption and distribution 9. While injection-based methods prove efficient for single delivery, prolonged and repeated injections often lead to body damage and chronic infection2.
Currently, there are two methods of IP injection in adult zerbafish4,10. However, both methods have limitations when delivering toxic compounds like doxorubicin, leading to severe damage to the body shape and fish mortality. The side effects can significantly complicate data interpretation. Although these challenges may be addressed with extensive training10, there is a clear need for a new IP injection method that minimizes side effects.
Here, our goal is to develop a new method of IP injection optimized for the effective delivery of doxorubicin into adult zebrafish, facilitating the generation of reliable doxorubicin-induced cardiomyopathy (DIC) models with minimized body damage and associated mortality.

作者聚焦： 斑马鱼心脏毒性模型的新型腹腔内注射方法

一种成年斑马鱼腹腔注射技术，可最大限度地减少身体损伤和相关死亡率

We aim to generate anthracycline induced cardiotoxicity models in adult zebrafish so that we can discover new therapeutic genes and compounds. A reliable IP injection method is essential to ensure the success of this project. So previously developed IP injection methods are the classic and alternative techniques.While the classic method penetrates the fish from the abdominal site through midline between the pelvic fins, the alternative method penetrates the fish from the dorsal site located between the pelvic and the anal fins. Owing to the toxicity of doxorubicin, significant body damage and mortality of the fish have been noted using the current two IP methods. New investigators need to be extensively trained before disease models can be reliably established in their hands.We designed a new IP injection method by adjusting the needle penetration location and the needle penetration angle. We inject through a natural hole located in the ventral region between pectoral fins and penetrate through a unique anterior to posterior direction.

成年斑马鱼体内改良腹腔注射阿霉素

首先，将 5 毫克阿霉素粉末溶解在 1 毫升蒸馏水中。将原液分装到 1.5 毫升试管中。接下来，将体重差异小于 10% 的鱼归为一组。用三卡因在胚胎水中麻醉鱼后，将鱼从水中取出。用干净的滤纸轻拍鱼体的两侧，以去除多余的水分。接下来，将麻醉的鱼放入嵌入海绵的凹槽中，腹部朝上。在配备 10 号针头的 34 微升注射器中加入 5 微升阿霉素工作溶液。从胸鳍中点向心脏腔后侧以近零度的角度插入针头。将针头对准尾巴，进入银色皮肤下方。缓慢分配 dox 溶液，然后沿原始路径抽出针头以防止泄漏。dox 溶液会将腹部染成红色。

modified-intraperitoneal-injection-of-doxorubicin-into-adult-zebrafish

Research

JoVE Journal

Medicine

Modified Intraperitoneal Injection of Doxorubicin into Adult Zebrafish

114 次观看.  Mayo Clinic. 首先，将 5 毫克阿霉素粉末溶解在 1 毫升蒸馏水中。将原液分装到 1.5 毫升试管中。接下来，将体重差异小于 10% 的鱼归为一组。用三卡因在胚胎水中麻醉鱼后，将鱼从水中取出。用干净的滤纸轻拍鱼体的两侧，以去除多余的水分。接下来，将麻醉的鱼放入嵌入海绵的凹槽中，腹部朝上。在配备 10 号针头的 34 微升注射器中加入 5 微升阿霉素工作溶液。从胸鳍中点向...

视频: 成年斑马鱼体内改良腹腔注射阿霉素

An Intraperitoneal Injection Technique in Adult Zebrafish that Minimizes Body Damage and Associated Mortality

The adult zebrafish (Danio rerio), which is genetically accessible, is being employed as a valuable vertebrate model to study human disorders such as cardiomyopathy. Intraperitoneal (IP) injection is an important method that delivers compounds to the body for either testing therapeutic effects or generating disease models such as doxorubicin-induced cardiomyopathy (DIC). Currently, there are two methods of IP injection. Both&#160;methods have limitations when handling toxic compounds such as doxorubicin, which result in side effects manifesting as severe damage to the body shape and fish death. While these shortcomings could be overcome by extensive investigator training, a new IP injection method that has minimal side effects is desirable. Here, a unique IP injection method that is able to handle toxic compounds is reported. Consistently reduced cardiac function can result without incurring significant fish death. The technique can be easily mastered by researchers who have minimal experience with adult zebrafish.

A new intraperitoneal (IP) injection method in adult zebrafish is described. When handling toxic compounds such as doxorubicin, this procedure is more effective than the two previously reported IP methods. The technique is designed to be easily adopted by researchers with limited experience in the zebrafish model.

The adult zebrafish (Danio rerio), which is genetically accessible, is being employed as a valuable vertebrate model to study human disorders such as ...