Application of Flow Vermimetry for Quantification and Analysis of the <em>Caenorhabditis elegans</em> Gut Microbiome

Fan Zhang; Dana Blackburn; Ciara N. Hosea; Adrien Assi&#233;; Buck S. Samuel

doi:10.3791/64605

Caenorhabditis elegans Bağırsak Mikrobiyomunun Miktar Tayini ve Analizi için Akış Vermim...

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Application of Flow Vermimetry for Quantification and Analysis of the Caenorhabditis elegans Gut Microbiome

Caenorhabditis elegans Bağırsak Mikrobiyomunun Miktar Tayini ve Analizi için Akış Vermimetrisinin Uygulanması

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08:38 min

March 31, 2023

DOI: 10.3791/64605-v

Fan Zhang*¹, Dana Blackburn*¹, Ciara N. Hosea^1,2, Adrien Assié¹, Buck S. Samuel^1,2

¹Alkek Center for Metagenomics and Microbiome Research and Department of Molecular Virology and Microbiology,Baylor College of Medicine, ²Development, Disease Models and Therapeutics Program, Graduate School of Biomedical Sciences,Baylor College of Medicine

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Overview

This study investigates the molecular determinants driving host-microbiome interactions using the model organism Caenorhabditis elegans. A high-throughput pipeline was developed to assess gut microbiome colonization levels in individual worms, enabling the correlation of these levels with the worm's physiological states.

Key Study Components

Research Area

Host-microbiome interactions
Model organism use in microbiology
High-throughput experimental approaches

Background

The gut microbiome plays crucial roles in shaping host physiology.
Understanding these interactions can provide insights into health and disease.
C. elegans serves as an effective model for studying these dynamics.

Methods Used

High-throughput assessment of microbial colonization in live C. elegans individuals.
Real-time monitoring of interactions using various fluorescence detection techniques.
Procedures for isolating and processing worms for analysis.

Main Results

Demonstrated differences in colonization levels based on microbial conditions.
Observed correlation between microbiome colonization and host physiological states like developmental stages.
Identified specific microbial effects on the physiological traits of C. elegans.

Conclusions

This study highlights the utility of C. elegans as a model for exploring host-microbiome interactions.
The findings have implications for understanding microbial influence on host biology and potential applications in health research.

Frequently Asked Questions

What is the significance of using C. elegans in microbiome research?

C. elegans provides a simple yet effective platform to study complex biological interactions at a single organism level.

How does the high-throughput pipeline work?

It allows for the assessment of microbiome colonization in live worms, linking it to physiological parameters.

What methodologies are employed in this research?

The protocol employs washing, centrifugation, and real-time fluorescent monitoring to analyze microbial colonization.

What are the implications of the findings?

The study provides insights that may inform health-related microbiome research and its effects on development.

Can these methods be applied to other organisms?

While designed for C. elegans, similar methodologies could potentially be adapted for other model organisms.

What are the critical variables in the study?

Microbial type and concentration, as well as developmental stages of the worms, are critical to the outcomes observed.

How does this research contribute to our understanding of gut microbiomes?

It elucidates the interactions between specific microbes and host physiology, advancing our knowledge of gut health.

Caenorhabditis elegans , konakçı-mikrobiyom etkileşimlerini yönlendiren moleküler belirleyicileri incelemek için güçlü bir modeldir. C. elegans fizyolojisinin temel yönleriyle birlikte bağırsak mikrobiyom kolonizasyonunun tek hayvan seviyelerinin profilini çıkaran yüksek verimli bir boru hattı sunuyoruz.

Bağırsak mikrobiyomu, konakçının fizyolojisini şekillendirmede önemli rol oynar. Bu protokol, özellikle, tek, hücresel hayvanlarda floresan bağırsak mikrop kolonizasyon seviyelerinin büyük ölçekte yüksek verimli değerlendirmesine izin verir. Bu yöntemin en büyük faydalarından biri, canlı hayvanların bağırsak mikrobiyomunu gerçek zamanlı olarak izleme yeteneğidir, bu da herhangi bir değişikliğin konakçının fizyolojisi ile tanımlanmasına ve korelasyonuna izin verir.

Prosedürü göstermek, laboratuvarımdan bir araştırma görevlisi olan Dana Blackburn olacak. Solucanları bakteri çimlerinden bir ila iki mililitre M9-TX ile yıkayarak başlayın. Solucanları sterilize edilmiş iki mililitrelik 96 kuyu derinliğinde bir plakaya aktarın.

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