Murine Fecal Isolation and Microbiota Transplantation

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

doi:10.3791/64310

Isolamento Fecal Murino e Transplante de Microbiota

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Murine Fecal Isolation and Microbiota Transplantation

Isolamento Fecal Murino e Transplante de Microbiota

Full Text

5,786 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.

O objetivo aqui é delinear um protocolo para investigar os mecanismos de disbiose na doença cardiovascular. Este artigo discute como coletar e transplantar amostras fecais murinas assepticamente, isolar intestinos e usar o método "Swiss-roll", seguido por técnicas de imunomarcação para interrogar alterações no trato gastrointestinal.

Aqui descrevemos um protocolo para orientar outras pessoas na investigação dos mecanismos de disbiose na doença, desde a coleta de amostras fecais até o uso do método Swiss roll para estudar alterações no intestino. Para começar, borrife o peito e as laterais do camundongo eutanasiado com etanol a 70% e abra cuidadosamente a pele e a cavidade peritoneal para expor o trato gastrointestinal. Isole o ceco e use tesoura cirúrgica estéril para cortá-lo ao meio.

Expor brevemente o ceco, e cortar 0,5 centímetros proximalmente do íleo e 0,5 centímetros distalmente em sua junção com o cólon. Transfira o ceco isolado para uma placa de Petri estéril. Use uma espátula estéril para transferir o conteúdo cecal para tubos estéreis e armazene as alíquotas em um freezer de 80 graus Celsius.

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