Murine Fecal Isolation and Microbiota Transplantation

Jeanne A. Ishimwe; Jianyong Zhong; Valentina Kon; Annet Kirabo

doi:10.3791/64310

Isolement fécal murin et transplantation de microbiote

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Murine Fecal Isolation and Microbiota Transplantation

Isolement fécal murin et transplantation de microbiote

Full Text

5,780 Views

07:32 min

May 26, 2023

DOI: 10.3791/64310-v

Jeanne A. Ishimwe¹, Jianyong Zhong^2,3, Valentina Kon², Annet Kirabo¹

¹Division of Clinical Pharmacology, Department of Medicine Vanderbilt University Medical Center and Department of Molecular Physiology and Biophysics,Vanderbilt University, ²Division of Pediatric Nephrology Department of Pediatrics,Vanderbilt University Medical Center, ³Department of Pathology, Microbiology, and Immunology,Vanderbilt University Medical Center

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Overview

This article outlines a protocol for investigating dysbiosis mechanisms in cardiovascular disease. It details the aseptic collection of murine fecal samples and the use of the Swiss-roll method for studying gastrointestinal changes.

Key Study Components

Area of Science

Neuroscience
Microbiology
Cardiovascular Research

Background

Dysbiosis refers to microbial imbalance in the gut.
It is linked to various diseases, including cardiovascular conditions.
Understanding gut microbiota's role can inform therapeutic strategies.
Protocols for studying dysbiosis are essential for advancing research.

Purpose of Study

To provide a detailed protocol for fecal sample collection.
To demonstrate the Swiss-roll method for gut analysis.
To facilitate research on dysbiosis in cardiovascular disease.

Methods Used

Aseptically collect murine fecal samples.
Isolate the gastrointestinal tract, focusing on the cecum.
Employ the Swiss-roll method for tissue analysis.
Utilize immunostaining techniques to observe changes.

Main Results

Successful collection and preservation of fecal samples.
Effective isolation of the cecum for analysis.
Implementation of the Swiss-roll method yielded insightful data.
Immunostaining revealed significant changes in gut morphology.

Conclusions

The outlined protocol is a valuable tool for studying dysbiosis.
Findings contribute to understanding gut health in cardiovascular disease.
Future research can build on these methods for deeper insights.

Frequently Asked Questions

What is dysbiosis?

Dysbiosis is an imbalance in the microbial communities in the gut, which can affect health.

Why is the Swiss-roll method used?

The Swiss-roll method allows for detailed examination of gut tissue architecture and microbiota interactions.

How are fecal samples collected?

Fecal samples are collected aseptically from euthanized mice to prevent contamination.

What techniques are used for analysis?

Immunostaining techniques are employed to visualize changes in gut tissue.

What are the implications of this research?

Understanding dysbiosis can lead to better therapeutic strategies for cardiovascular diseases.

Can this protocol be applied to other diseases?

Yes, the protocol can be adapted to study dysbiosis in various diseases beyond cardiovascular conditions.

L’objectif ici est de définir un protocole pour étudier les mécanismes de la dysbiose dans les maladies cardiovasculaires. Cet article explique comment collecter et transplanter de manière aseptique des échantillons fécaux murins, isoler les intestins et utiliser la méthode « Swiss-roll », suivie de techniques d’immunomarquage pour interroger les changements dans le tractus gastro-intestinal.

Ici, nous avons décrit un protocole pour guider les autres dans l’étude des mécanismes de la dysbiose dans la maladie, de la collecte d’échantillons fécaux à l’utilisation de la méthode suisse du rouleau pour étudier les changements dans l’intestin. Pour commencer, vaporisez la poitrine et les côtés de la souris euthanasiée avec de l’éthanol à 70% et ouvrez soigneusement la peau et la cavité péritonéale pour exposer le tractus gastro-intestinal. Isolez le caecum et utilisez des ciseaux chirurgicaux stériles pour le couper en deux.

Exposez brièvement le caecum et coupez 0,5 centimètre à proximité de l’iléon et 0,5 centimètre distalement à sa jonction avec le côlon. Transférer le caecum isolé sur une boîte de Petri stérile. Utilisez une spatule stérile pour transférer le contenu de la cæcale dans des tubes stériles et conservez les aliquotes dans un congélateur à 80 degrés Celsius.

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