modified intraperitoneal injection of doxorubicin into adult zebrafish

Inyección intraperitoneal optimizada para la administración de compuestos en pez cebra adulto

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.

Autor destacado: Nuevo método de inyección intraperitoneal para modelos de cardiotoxicidad del pez cebra

Una técnica de inyección intraperitoneal en peces cebra adultos que minimiza el daño corporal y la mortalidad asociada

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.

Inyección intraperitoneal modificada de doxorrubicina en pez cebra adulto

Para empezar, disuelve cinco miligramos de doxorrubicina en polvo en un mililitro de agua destilada. Alícuota el caldo en tubos de 1,5 mililitros. A continuación, agrupe juntos los peces con una diferencia de peso corporal de menos del 10%.Después de anestesiar a los peces en agua embrionaria con tricaína, saque a los peces del agua. Frota ambos lados del cuerpo del pez con papel de filtro limpio para eliminar el exceso de agua. A continuación, coloque el pez anestesiado en la ranura de una esponja incrustada con el abdomen hacia arriba.Llene una jeringa de 10 microlitros equipada con una aguja de calibre 34 con cinco microlitros de solución de trabajo de doxorrubicina. Inserte la aguja en un ángulo de casi cero grados desde el punto medio de la aleta pectoral hacia el lado posterior de la cavidad cardíaca. Dirige la aguja hacia la cola y pasa por debajo de la piel plateada.Dispense lentamente la solución de dox, luego retire la aguja a lo largo de la trayectoria original para evitar fugas. La solución dox coloreará el abdomen de rojo.

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

Research

JoVE Journal

Medicine

Modified Intraperitoneal Injection of Doxorubicin into Adult Zebrafish

114 vistas.  Mayo Clinic. Para empezar, disuelve cinco miligramos de doxorrubicina en polvo en un mililitro de agua destilada. Alícuota el caldo en tubos de 1,5 mililitros. A continuación, agrupe juntos los peces con una diferencia de peso corporal de menos del 10%.Después de anestesiar a los peces en agua embrionaria con tricaína, saque a los peces del agua. Frota ambos lados del cuerpo del pez con papel de filtro limpio para eliminar el exceso de agua. A continuación, coloque el pez anestesiado en la ran...

Video: Inyección intraperitoneal modificada de doxorrubicina en pez cebra adulto

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 ...