Brain Ventricular Microinjections of Lipopolysaccharide into Larval Zebrafish to Assess Neuroinflammation and Neurotoxicity

Yulin He; Simon Ming Yuen Lee

doi:10.3791/64313

Microinjections ventriculaires cérébrales de lipopolysaccharide dans des larves de poisson-zèbre ...

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Neuroscience

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JoVE Journal Neuroscience

Brain Ventricular Microinjections of Lipopolysaccharide into Larval Zebrafish to Assess Neuroinflammation and Neurotoxicity

Microinjections ventriculaires cérébrales de lipopolysaccharide dans des larves de poisson-zèbre pour évaluer la neuroinflammation et la neurotoxicité

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07:31 min

August 23, 2022

DOI: 10.3791/64313-v

Yulin He¹, Simon Ming Yuen Lee^1,2

¹State Key Laboratory of Quality Research in Chinese Medicine and Institute of Chinese Medical Sciences,University of Macau, Avenida da Universidade, ²Department of Pharmaceutical Sciences, Faculty of Health Sciences,University of Macau, Avenida da Universidade

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Overview

This study presents a protocol for microinjecting lipopolysaccharide into the brain ventricular region of zebrafish larvae to investigate neuroinflammatory responses and associated neurotoxicity. The model is aimed at understanding the pathological changes in the brain and evaluating potential anti-neuroinflammatory drugs using in vivo imaging techniques.

Key Study Components

Area of Science

Neuroscience
Neuroinflammation
Zebrafish models

Background

Neuroinflammation plays a pivotal role in various neurological disorders.
Research aims to develop alternative in vivo models to study disease progression.
The zebrafish model offers advantages for real-time imaging of the brain.
This technique facilitates the evaluation of neuroinflammatory drugs quickly and effectively.

Purpose of Study

To assess the neuroinflammatory response following lipopolysaccharide injection.
To investigate neurotoxic effects resulting from inflammatory processes.
To provide insights into the efficacy of potential therapeutic compounds.

Methods Used

The study utilizes microinjection of solutions into zebrafish larval brains.
Zebrafish larvae are subjected to lipopolysaccharide injection to assess neuroinflammation.
Behavioral assays and real-time PCR are employed to monitor responses.
Critical protocols include needle preparation, larval positioning, and injection techniques.
Microscopy and image analysis are used to observe outcomes post-injection.

Main Results

The injection of lipopolysaccharide resulted in significant neurotoxic effects, including loss of specific neurons.
Increased expression of pro-inflammatory cytokines and signs of neuroinflammation were observed.
Behavioral changes indicated locomotion deficiencies, correlating with the level of lipopolysaccharide administered.
Neutrophil recruitment to the brain region was markedly increased in response to treatment.

Conclusions

This study establishes a reliable method for investigating neuroinflammation in zebrafish.
It highlights the potential for this approach in testing new anti-inflammatory drugs.
The findings contribute to the understanding of mechanisms underlying neuroinflammation and related disorders.

Frequently Asked Questions

What are the advantages of using zebrafish larvae for this study?

Zebrafish larvae offer transparency for real-time imaging and a rapid developmental timeline, making them ideal for in vivo studies of neuroinflammation.

How is the lipopolysaccharide injection administered?

Lipopolysaccharide is injected into the brain ventricular region using a microinjection apparatus, following a precise setup and technique to ensure accurate dosing.

What types of outcomes are assessed post-injection?

Outcomes include neuronal loss, behavioral changes, and gene expression levels associated with neuroinflammation.

What are the key limitations of this model?

One limitation is that the responses in zebrafish may not fully replicate those in mammals, potentially affecting the translational value of the findings.

Can this method be adapted for other types of injections?

Yes, this microinjection technique can be adapted for various substances or experimental conditions in zebrafish larvae to study different biological processes.

How are behavioral assays conducted in this study?

Behavioral assays are performed using an automated video tracking system to monitor movement patterns of the larvae after treatment.

Ce protocole démontre la micro-injection de lipopolysaccharide dans la région ventriculaire du cerveau dans un modèle larvaire de poisson zèbre pour étudier la réponse neuroinflammatoire et la neurotoxicité qui en résultent.

La neuroinflammation est un acteur clé dans divers troubles neurologiques. Par conséquent, il est d’un grand intérêt de rechercher et de développer des modèles alternatifs de neuroinflammation in vivo pour faciliter les études du développement pathologique. La technique décrite ici est un excellent outil pour mieux comprendre les changements pathologiques dans le cerveau via l’imagerie in vivo, et pour évaluer rapidement et efficacement les anti-neuro-inflammatoires possibles.

Préparez-vous à l’injection en tirant les capillaires en verre à l’aide d’un extracteur de micropipettes en suivant le protocole en cinq étapes pour le tirage du tube capillaire en verre. Ouvrez la pointe de l’aiguille à la taille appropriée à un angle à l’aide d’une pince. Remplissez l’aiguille avec 0,1 millilitre d’huile minérale pour vous assurer qu’il n’y a pas de bulles.

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Neuroscience numéro 186 Cerveau micro-injection ventriculaire poisson zèbre lipopolysaccharide neuroinflammation neurotoxicité