Following Endocardial Tissue Movements via Cell Photoconversion in the Zebrafish Embryo

Renee Wei-Yan Chow; Paola Lamperti; Emily Steed; Francesco Boselli; Julien Vermot

doi:10.3791/57290

Mouvements de tissu endocardique suivant via Photoconversion de cellules dans l’embryon de poisso...

JoVE Journal
Developmental Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Developmental Biology

Following Endocardial Tissue Movements via Cell Photoconversion in the Zebrafish Embryo

Mouvements de tissu endocardique suivant via Photoconversion de cellules dans l’embryon de poisson zèbre

Full Text

6,949 Views

09:38 min

February 20, 2018

DOI: 10.3791/57290-v

Renee Wei-Yan Chow*^1,2,3,4, Paola Lamperti*^1,2,3,4, Emily Steed^1,2,3,4, Francesco Boselli^1,2,3,4, Julien Vermot^1,2,3,4

¹Institut de Génétique et de Biologie Moléculaire et Cellulaire, ²UMR7104,Centre National de la Recherche Scientifique, ³U964,Institut National de la Santé et de la Recherche Médicale, ⁴Université de Strasbourg

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol describes a method for tracking endocardial cells during the development of the atrioventricular canal and heart valves in living zebrafish embryos. The technique overcomes challenges associated with traditional time-lapse microscopy due to the rapid heartbeat of the embryos.

Key Study Components

Area of Science

Cardiac development
Cell tracking
Fluorescent protein photoconversion

Background

Endocardial cells play a crucial role in heart development.
Tracking these cells is essential for understanding cardiac morphogenesis.
Traditional imaging methods are limited by the heart's rapid movement.
This method provides a solution to visualize cell behavior in real-time.

Purpose of Study

To follow endocardial cells during atrioventricular canal and valve formation.
To investigate cell behavior in various genetic mutants.
To enhance understanding of cardiac development mechanisms.

Methods Used

Photoconversion of Kaede fluorescent protein.
Live imaging of zebrafish embryos.
Tracking of endocardial cells during heart development.
Utilization of advanced microscopy techniques.

Main Results

Successful tracking of endocardial cells in live embryos.
Insights into cell behavior during heart valve formation.
Demonstration of the method's effectiveness in overcoming imaging challenges.
Potential applications in studying cardiac development in mutants.

Conclusions

The method allows for detailed observation of endocardial cells.
It provides a valuable tool for researchers studying cardiac development.
Future studies can leverage this technique to explore various cardiac conditions.

Frequently Asked Questions

What is the main advantage of this technique?

It allows for tracking endocardial cells despite the rapid heartbeat of zebrafish embryos, which is challenging with traditional methods.

Who demonstrates the procedure?

Renee, a postdoc in the lab, demonstrates the procedure.

What type of cells are being tracked?

Endocardial cells during atrioventricular canal and heart valve formation.

How does this method contribute to cardiac research?

It helps researchers understand cell behavior in different genetic backgrounds, enhancing knowledge of cardiac development.

Is this method applicable to other species?

While this study focuses on zebrafish, similar techniques may be adapted for other model organisms.

What is the significance of the Kaede fluorescent protein?

Kaede allows for photoconversion, enabling researchers to track specific cells over time in live embryos.

Ce protocole décrit une méthode pour la photoconversion de la protéine fluorescente de Kaede dans les cellules du vivant endocardiques embryon de poisson zèbre qui permet le suivi des cellules endocardiaques pendant le canal atrio-ventriculaire et le développement de valve auriculo-ventriculaire cardiaque .

L’objectif global de cette procédure est de suivre les cellules endocardiques pendant la formation du canal auriculo-ventriculaire et des valves cardiaques. Cette méthode peut aider à répondre à des questions clés dans le domaine du développement cardiaque, telles que comprendre comment les cellules endocardiques se comportent chez différents types de mutants. Le suivi des cellules pendant la formation de l’AVC et de la valve est difficile en raison du battement rapide du cœur.

Il est difficile de suivre les cellules via la microscopie time-lapse traditionnelle. Le principal avantage de cette technique est qu’elle contourne certaines de ces difficultés. La démonstration de la procédure sera assurée par Renee, postdoctorante dans mon laboratoire.

View the full transcript and gain access to thousands of scientific videos