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

Toxicidad del amiloide β42 de modelado y la neurodegeneración en cerebro de pez cebra adulto

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Neuroscience

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JoVE Journal Neuroscience

Modeling Amyloid-β42 Toxicity and Neurodegeneration in Adult Zebrafish Brain

Toxicidad del amiloide β42 de modelado y la neurodegeneración en cerebro de pez cebra adulto

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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 describe la síntesis, caracterización y la inyección de péptidos monomérico amiloide-β42 para generar toxicidad amiloide en pez cebra adulto para establecer un modelo de la enfermedad de Alzheimer, seguido por análisis histológicos y la detección de agregaciones.

El objetivo general de este método es sintetizar péptidos de beta amiloide 42 e inyectarlos en el cerebro del pez cebra adulto, para generar un modelo particular de dosis de amiloide como se observa en la enfermedad de Alzheimer. Este método puede ayudarnos a abordar cuestiones importantes en el campo de la neurociencia, como el efecto de la toxicidad del amiloide en las células madre neurales, y cómo podemos provocar la respuesta de rejuvenecimiento en condiciones tan perjudiciales. Las principales ventajas de esta técnica son su reporte, su reproducibilidad y los fenotipos que obtenemos recuerdan mucho a la enfermedad de Alzheimer en el cerebro humano.

Las implicaciones de esta técnica se extienden hacia la terapia de la enfermedad de Alzheimer ya que proporciona un modelo natural de vertebrado, que es capaz de activar las células madre neurales en la enfermedad, por lo que el pez cebra puede enseñarnos cómo recuperar la neurodegeneración. Para preparar el péptido beta amiloide 42, cargue un sintetizador de péptidos automatizado con 500 miligramos de resina protegida contra FMOC como fase sólida. A continuación, cargue 0,5 aminoácidos molares disueltos protegidos contra FMOC en el volumen requerido, según lo calculado para el sintetizador respectivo.

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