Modeling Amyloid-&#946;42 Toxicity and Neurodegeneration in Adult Zebrafish Brain

Prabesh Bhattarai; Alvin Kuriakose Thomas; Mehmet Ilyas Cosacak; Christos Papadimitriou; Violeta Mashkaryan; Yixin Zhang; Caghan Kizil

doi:10.3791/56014

Toxicidade do Amyloid-β42 de modelagem e neurodegeneração no cérebro adulto de Zebrafish

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Neuroscience

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JoVE Journal Neuroscience

Modeling Amyloid-β42 Toxicity and Neurodegeneration in Adult Zebrafish Brain

Toxicidade do Amyloid-β42 de modelagem e neurodegeneração no cérebro adulto de Zebrafish

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10:01 min

October 25, 2017

DOI: 10.3791/56014-v

Prabesh Bhattarai¹, Alvin Kuriakose Thomas², Mehmet Ilyas Cosacak¹, Christos Papadimitriou¹, Violeta Mashkaryan³, Yixin Zhang², Caghan Kizil^1,3

¹German Centre for Neurodegenerative Diseases (DZNE) Dresden within Helmholtz Association, ²B CUBE, Center for Molecular Bioengineering,Technische Universität Dresden, ³Center for Regenerative Therapies Dresden (CRTD),TU Dresden

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Overview

This protocol outlines the synthesis and injection of amyloid-Î²42 peptides into adult zebrafish to create an Alzheimer's disease model. The study aims to explore amyloid toxicity effects on neural stem cells and potential rejuvenation responses.

Key Study Components

Area of Science

Neuroscience
Alzheimer's Disease Research
Animal Models

Background

Amyloid-Î²42 peptides are implicated in Alzheimer's disease pathology.
Zebrafish serve as a natural vertebrate model for studying neurodegeneration.
Understanding amyloid toxicity can inform therapeutic strategies.
This method allows for the investigation of neural stem cell responses in disease conditions.

Purpose of Study

To synthesize amyloid-Î²42 peptides for injection into zebrafish.
To establish a model that mimics Alzheimer's disease.
To investigate the effects of amyloid toxicity on neural stem cells.

Methods Used

Synthesis of amyloid-Î²42 peptides using an automated peptide synthesizer.
Injection of synthesized peptides into the adult zebrafish brain.
Histological analyses to detect amyloid aggregations.
Assessment of neural stem cell activation in response to amyloid toxicity.

Main Results

Successful synthesis and injection of amyloid-Î²42 peptides.
Observation of amyloid toxicity effects on zebrafish neural stem cells.
Phenotypes observed are reminiscent of human Alzheimer's disease.
Potential for zebrafish to inform therapeutic approaches for neurodegeneration.

Conclusions

The method provides a reproducible model for studying Alzheimer's disease.
Zebrafish can activate neural stem cells under amyloid toxicity conditions.
This research may lead to insights into neurodegeneration recovery strategies.

Frequently Asked Questions

What is the significance of using zebrafish in this study?

Zebrafish are a natural vertebrate model that allows for the observation of neurodegeneration and the activation of neural stem cells in response to amyloid toxicity.

How are amyloid-Î²42 peptides synthesized?

Amyloid-Î²42 peptides are synthesized using an automated peptide synthesizer with FMOC protected amino acids.

What are the main advantages of this technique?

The technique is reproducible, provides relevant phenotypes, and helps in understanding the effects of amyloid toxicity on neural stem cells.

What implications does this research have for Alzheimer's therapy?

This research may inform therapeutic strategies by revealing how to activate neural stem cells in conditions of amyloid toxicity.

What histological analyses are performed in this study?

Histological analyses are conducted to detect amyloid aggregations in the zebrafish brain post-injection.

Este protocolo descreve a síntese, caracterização e injeção de peptídeos monomérico amiloide-β42 para gerar toxicidade amiloide no zebrafish adulto para estabelecer um modelo de doença de Alzheimer, seguido de análise histológica e detecção de agregações.

O objetivo geral deste método é sintetizar peptídeos beta 42 amilóides e injetá-los no cérebro adulto do peixe-zebra, para gerar um modelo particular de doses de amilóide, como visto na doença de Alzheimer. Este método pode nos ajudar a abordar questões importantes no campo da neurociência, como o efeito da toxicidade amilóide nas células-tronco neurais e como podemos provocar a resposta de rejuvenescimento em tais condições prejudiciais. As principais vantagens desta técnica são o seu relatório, é reprodutível e os fenótipos que obtemos são altamente reminiscentes da doença de Alzheimer em cérebros humanos.

As implicações desta técnica se estendem à terapia da doença de Alzheimer porque fornece um modelo natural de vertebrado, que é capaz de ativar células-tronco neurais em condições de doença, portanto, o peixe-zebra pode nos ensinar como recuperar a neurodegeneração. Para preparar o peptídeo beta amilóide 42, carregue um sintetizador de peptídeos automatizado com 500 miligramas de resina protegida por FMOC como fase sólida. Em seguida, carregue 0,5 aminoácidos protegidos por FMOC molar dissolvidos no volume necessário, conforme calculado para o respectivo sintetizador.

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