Optogenetic Inhibition of Rho1-Mediated Actomyosin Contractility Coupled with Measurement of Epithelial Tension in <em>Drosophila</em> Embryos

Hanqing Guo; Michael Swan; Bing He

doi:10.3791/65314

Inibizione optogenetica della contrattilità dell'actomiosina mediata da Rho1 accoppiata alla misu...

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Developmental Biology

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JoVE Journal Developmental Biology

Optogenetic Inhibition of Rho1-Mediated Actomyosin Contractility Coupled with Measurement of Epithelial Tension in Drosophila Embryos

Inibizione optogenetica della contrattilità dell'actomiosina mediata da Rho1 accoppiata alla misurazione della tensione epiteliale negli embrioni di Drosophila

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12:35 min

April 14, 2023

DOI: 10.3791/65314-v

Hanqing Guo^1,2, Michael Swan³, Bing He¹

¹Department of Biological Sciences,Dartmouth College, ²School of Life Sciences,Westlake University, ³Department of Molecular Biology,Princeton University

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Overview

This study investigates the role of actomyosin contractility in tissue morphogenesis, particularly focusing on Drosophila embryos. The research employs an optogenetic system to rapidly inhibit Rho1-mediated actomyosin contractility, allowing for the observation of immediate changes in epithelial tension.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Developmental Biology

Background

Actomyosin contractility is crucial for the formation of complex tissue structures.
Understanding the mechanical forces involved in morphogenesis is essential for developmental biology.
Conventional genetic approaches are limited in their ability to manipulate actomyosin contractility in vivo.
This study aims to provide a method for acute inactivation of actomyosin contractility.

Purpose of Study

To explore how actomyosin contractility influences epithelial folding.
To develop a rapid manipulation technique for studying tissue behavior.
To enhance understanding of the genetic processes regulating morphogenesis.

Methods Used

Optogenetic system for inactivation of Rho1-mediated contractility.
In vivo experiments using Drosophila embryos.
Measurement of epithelial tension changes post-inactivation.
Analysis of tissue behavior and properties following manipulation.

Main Results

Immediate loss of epithelial tension was observed upon actomyosin inactivation.
The optogenetic approach allowed for precise temporal control of contractility.
Findings contribute to understanding the mechanics of tissue morphogenesis.
This method can be applied to study other genetic processes in development.

Conclusions

Actomyosin contractility is a key regulator of epithelial tension and tissue structure.
The optogenetic system provides a valuable tool for developmental biology research.
Future studies can leverage this approach to further investigate morphogenetic mechanisms.

Frequently Asked Questions

What is actomyosin contractility?

Actomyosin contractility refers to the contractile forces generated by the interaction of actin filaments and nonmuscle myosin II, which are crucial for tissue morphogenesis.

How does the optogenetic system work?

The optogenetic system allows for the rapid and precise inactivation of specific proteins, such as Rho1, using light to control cellular processes in real-time.

Why is Drosophila used in this study?

Drosophila embryos are a well-established model for studying developmental processes and allow for genetic manipulation and observation of tissue behavior.

What are the implications of this research?

This research enhances our understanding of the mechanical forces driving tissue morphogenesis and provides a new tool for studying genetic processes in development.

Can this method be applied to other organisms?

While this study focuses on Drosophila, the optogenetic approach may be adapted for use in other model organisms to study similar processes.

La contrattilità dell'actomiosina svolge un ruolo importante nella morfogenesi cellulare e tissutale. Tuttavia, è difficile manipolare la contrattilità dell'actomiosina in vivo in modo acuto. Questo protocollo descrive un sistema optogenetico che inibisce rapidamente la contrattilità dell'actomiosina mediata da Rho1 negli embrioni di Drosophila , rivelando l'immediata perdita di tensione epiteliale dopo l'inattivazione dell'actomiosina in vivo.

La nostra ricerca studia la morfogenesi tissutale, la formazione di complesse strutture tissutali tridimensionali in fase di sviluppo. Siamo interessati ai geni e alle molecole che regolano la morfogenesi e cerchiamo di comprendere i principi fisici alla base della morfogenesi. Ad esempio, come vengono generate le forze meccaniche e come guidano la riabilitazione dei tessuti.

Le forze contrattili generate dall'actina filamentosa e dalla miosina II non muscolare, nota anche come contrattilità dell'actomiosina, sono una delle forze più importanti che guidano la morfogenesi dei tessuti. La nostra ricerca attuale affronta il modo in cui la contrattilità dell'actomiosina media il ripiegamento dei fogli cellulari epiteliali del sangue, un meccanismo fondamentale di costruzione dei tessuti nello sviluppo. Una comprensione approfondita del ruolo della contrattilità dell'actomiosina nel ripiegamento epiteliale e in altri processi morfogenetici richiede approcci in grado di inattivare rapidamente l'actomiosina nel momento e nel luogo designati e registra l'impatto immediato del comportamento e delle proprietà dei tessuti.

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