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JoVE Journal
Developmental Biology
クロストリジオイド・ディフィシルの幼虫ゼブラフィッシュ感染モデルの開発
クロストリジオイド・ディフィシルの幼虫ゼブラフィッシュ感染モデルの開発
JoVE Journal
Developmental Biology
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JoVE Journal Developmental Biology
Development of a Larval Zebrafish Infection Model for Clostridioides difficile

クロストリジオイド・ディフィシルの幼虫ゼブラフィッシュ感染モデルの開発

Full Text
6,983 Views
09:13 min
February 14, 2020

DOI: 10.3791/60793-v

Junkai Li1, Can M. Ünal2, Kazuhiko Namikawa1, Michael Steinert3,4,5, Reinhard W. Köster1

1Division of Cellular and Molecular Neurobiology, Zoological Institute,Technische Universität Braunschweig, 2Department of Molecular Biotechnology,Turkish-German University, 3Institut für Mikrobiologie,Technische Universität Braunschweig, 4Braunschweig Integrated Centre of Systems Biology, 5Helmholtz Centre for Infection Research

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Overview

This article presents a safe and effective method for infecting zebrafish larvae with fluorescently labeled anaerobic C. difficile through microinjection and noninvasive microgavage. This technique allows researchers to investigate the innate immune response to Clostridium infection.

Key Study Components

Area of Science

  • Neuroscience
  • Microbiology
  • Immunology

Background

  • Understanding the interaction between the innate immune system and pathogens is crucial.
  • Macrophages play a key role in recognizing and phagocytosing pathogens.
  • Current methods for studying these interactions can be limited.
  • This method offers a more representative model of human infection.

Purpose of Study

  • To explore how the innate immune system responds to Clostridium difficile.
  • To determine the mechanisms of macrophage recognition and phagocytosis of this pathogen.
  • To develop a versatile method applicable to various anaerobic bacteria.

Methods Used

  • Microinjection of zebrafish larvae with fluorescently labeled C. difficile.
  • Noninvasive microgavage technique for oral administration.
  • Measurement of bacterial culture density using OD600.
  • Washing and resuspension of bacterial cultures in PBS.

Main Results

  • The method allows manipulation of infection time.
  • It closely mimics natural human infection routes.
  • Facilitates the study of immune responses in a live model.
  • Can be adapted for various anaerobic bacterial studies.

Conclusions

  • This method provides a novel approach to studying Clostridium infections.
  • It enhances understanding of immune system interactions with pathogens.
  • The versatility of the method opens new avenues for research.

Frequently Asked Questions

What is the significance of using zebrafish larvae in this study?
Zebrafish larvae provide a live model to study immune responses in real-time, closely mimicking human infection processes.
How does this method compare to traditional infection models?
This method is less invasive and allows for manipulation of infection timing, providing a more accurate representation of human infections.
What are the advantages of using fluorescently labeled bacteria?
Fluorescent labeling allows for easy visualization and tracking of bacterial interactions with host immune cells.
Can this method be used for other pathogens?
Yes, the method is versatile and can be adapted for various anaerobic bacteria.
What are the implications of this research?
The findings could lead to better understanding and treatment of Clostridium infections and similar diseases.

ここで提示された、ゼブラフィッシュ幼虫を蛍光標識された嫌気性C.ディフィシルでマイクロインジェクションおよび非侵襲的なマイクロガベージによって感染させる安全で効果的な方法です。

この方法は、マクロファージがこの病原体を認識または貪食する場合や食作用方法など、クロストリジウム感染における自然免疫系の役割に関する重要な質問に答えることができます。この方法の主な利点は、多くの異なる嫌気性細菌に適用できること、感染時間を操作できること、および経口投与が正常なヒト感染をはるかに近く表していることである。培養後、培養液を1ミリリットル分光光度計キュベットに移し、OD600を測定する。

最終体積の1ミリリットルで1〜1.2の最終ODに到達するために、必要な量を新鮮な1.5ミリリットルのチューブに移します。1回1ミリリットルの1回PBSで洗浄し、遠心分離機を5,000倍gで室温で3分間洗浄します。1X PBSの1ミリリットルで再中断します。

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発生生物学 問題156 ゼブラフィッシュ ダニオ レリオ クロストリジオイデス ディフィシル感染 マイクロインジェクション 生染色 マイクロガバデー 嫌気性 解剖 グノトビオティックゼブラフィッシュ

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