modified intraperitoneal injection of doxorubicin into adult zebrafish

성인 제브라피쉬의 화합물 전달을 위해 최적화된 복강내 주사

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.

저자 스포트라이트: 제브라피시 심장 독성 모델을 위한 새로운 복강 내 주사 방법

성체 제브라피시의 복강 내 주사 기법으로 신체 손상 및 관련 폐사율을 최소화합니다.

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.

성인 제브라피쉬에 대한 독소루비신의 수정된 복강내 주사

먼저 독소루비신 가루 5mg을 증류수 1ml에 녹입니다. 스톡을 1.5밀리리터 튜브에 분취합니다. 다음으로, 체중 차이가 10% 미만인 물고기를 함께 그룹화합니다.트리카인으로 배아 물에 물고기를 마취 한 후 물고기를 물에서 꺼냅니다. 깨끗한 여과지로 물고기 몸의 양쪽을 두드려 과도한 물을 제거합니다. 다음으로, 마취된 물고기를 복부가 위를 향하도록 하여 내장된 스폰지의 홈에 놓습니다.34 게이지 바늘이 장착된 10마이크로리터 주사기에 5마이크로리터의 독소루비신 작업 용액을 채웁니다. 가슴 지느러미의 중간 지점에서 심강의 뒤쪽을 향해 거의 0도 각도로 바늘을 삽입합니다. 바늘을 꼬리 쪽으로 향하게 하고 은색 피부 아래로 이동합니다.dox 용액을 천천히 분배한 다음 누출을 방지하기 위해 원래 경로를 따라 바늘을 빼냅니다. 독스 용액은 복부를 붉게 물들입니다.

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

Research

JoVE Journal

Medicine

Modified Intraperitoneal Injection of Doxorubicin into Adult Zebrafish

114 조회수.  Mayo Clinic. 먼저 독소루비신 가루 5mg을 증류수 1ml에 녹입니다. 스톡을 1.5밀리리터 튜브에 분취합니다. 다음으로, 체중 차이가 10% 미만인 물고기를 함께 그룹화합니다.트리카인으로 배아 물에 물고기를 마취 한 후 물고기를 물에서 꺼냅니다. 깨끗한 여과지로 물고기 몸의 양쪽을 두드려 과도한 물을 제거합니다. 다음으로, 마취된 물고기를 복부가 위를 향하도록 하여 내장된 스폰지...

비디오: 성인 제브라피쉬에 대한 독소루비신의 수정된 복강내 주사

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 ...