Nephrotoxin Microinjection in Zebrafish to Model Acute Kidney Injury

Robert A. McKee; Rebecca A. Wingert

doi:10.3791/54241

Nefrotoxina Microinyección en pez cebra para modelar la lesión renal aguda

JoVE Journal
Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

Nephrotoxin Microinjection in Zebrafish to Model Acute Kidney Injury

Nefrotoxina Microinyección en pez cebra para modelar la lesión renal aguda

Full Text

9,322 Views

07:58 min

July 17, 2016

DOI: 10.3791/54241-v

Robert A. McKee^1,2, Rebecca A. Wingert^1,2

¹Center for Zebrafish Research, Department of Biological Sciences,University of Notre Dame, ²Center for Stem Cells and Regenerative Medicine, Department of Biological Sciences,University of Notre Dame

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol demonstrates the use of zebrafish as a model for studying renal injuries caused by nephrotoxins, such as gentamicin and cisplatin. The zebrafish model allows for real-time imaging of cell damage and regeneration following acute kidney injury.

Key Study Components

Area of Science

Nephrology
Pharmacology
Model Organisms

Background

Renal injuries from nephrotoxins can lead to complex disorders.
The pathogenesis of these disorders is not fully understood.
Zebrafish provide a unique opportunity for disease modeling.
Real-time imaging capabilities enhance the study of kidney injury.

Purpose of Study

To model acute kidney injury using zebrafish.
To observe the effects of nephrotoxins on renal function.
To identify potential renoprotective measures.

Methods Used

Preparation of embryo manipulation tools for microinjection.
Creation of fine tapered microinjection needles.
Positioning of embryos for injection.
Real-time imaging of cell damage and regeneration.

Main Results

Demonstrated the feasibility of using zebrafish for nephrotoxin studies.
Provided insights into the regeneration process post-injury.
Highlighted the advantages of real-time imaging in research.
Facilitated the identification of potential therapeutic approaches.

Conclusions

Zebrafish are a valuable model for studying nephrotoxicity.
Real-time imaging can enhance understanding of kidney injury mechanisms.
This approach may lead to the development of renoprotective strategies.

Frequently Asked Questions

What are nephrotoxins?

Nephrotoxins are substances that can cause damage to the kidneys, including certain drugs and chemicals.

Why use zebrafish for kidney injury studies?

Zebrafish allow for real-time imaging and have similar kidney structures to humans, making them ideal for modeling kidney injuries.

What is the significance of real-time imaging?

Real-time imaging enables researchers to observe the dynamic processes of cell damage and regeneration as they occur.

How can this research impact clinical practices?

Findings may lead to new therapeutic strategies to protect against nephrotoxic damage in patients.

What are common nephrotoxins studied?

Common nephrotoxins include antibiotics like gentamicin and chemotherapeutic agents like cisplatin.

What are the next steps in this research?

Future studies may focus on identifying specific renoprotective measures and further understanding the mechanisms of injury and repair.

Las lesiones renales efectuados desde nefrotóxicos, que incluyen medicamentos que van desde antibióticos a los agentes quimioterapéuticos, pueden dar lugar a trastornos complejos cuya patogénesis se conoce por completo. Este protocolo se muestra cómo el pez cebra se puede utilizar para el modelado de la enfermedad de estas condiciones, que se pueden aplicar a la identificación de medidas renoprotective.

El objetivo general de este protocolo es utilizar el pez cebra para observar el efecto de las nefrotoxinas como la gentamicina o el cisplatino para modelar la lesión renal aguda. La principal ventaja de esta técnica es que el modelo de pez cebra permite obtener imágenes en tiempo real del daño celular y la regeneración después de una lesión renal aguda. Para comenzar este procedimiento, prepare una herramienta de manipulación de embriones para posicionar los embriones para la microinyección colocando una punta de carga de gel en el extremo de una aguja de disección recta de cinco pulgadas y asegúrela con cinta adhesiva o superpegamento.

A continuación, prepare unas agujas de microinyección cónicas finas tirando de los vidrios de borosilicato pulidos al fuego con filamento utilizando un extractor de pipetas. Luego, use una hoja de afeitar o un filo de pinza fina para cortar las puntas de la aguja y crear puntas de inyección angulares afiladas. Posteriormente, coloque las agujas en una tira de arcilla para modelar en una placa de Petri para proteger las puntas de las agujas.

View the full transcript and gain access to thousands of scientific videos