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

Optogenetic Inhibition of Rho1-Mediated Actomyosin Contractility Coupled with Measurement of Epit...

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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

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

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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

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.

Actomyosin contractility plays an important role in cell and tissue morphogenesis. However, it is challenging to manipulate actomyosin contractility in vivo acutely. This protocol describes an optogenetic system that rapidly inhibits Rho1-mediated actomyosin contractility in Drosophila embryos, revealing the immediate loss of epithelial tension after the inactivation of actomyosin in vivo.

Our research studies tissue morphogenesis, the formation of complex three-dimensional tissue structures in development. We are interested in the genes and the molecules that regulate morphogenesis and seek to understand the physical principles underlying morphogenesis, for example, how mechanical forces are generated and how they drive tissue revitalization. Contractile forces generated by filamentous actin and nonmuscle myosin II, also known as actomyosin contractivity, is one of the most important forces that drive tissue morphogenesis.

Our current research addresses how actomyosin contractivity mediates the folding of blood epithelial cell sheets, a fundamental tissue construction mechanism in development. An in-depth understanding of the role of actomyosin contractivity in epithelial folding and answerable for genetic processes requires approaches that can quickly inactivate actomyosin at mass limited time and location and record the immediate impact of tissue behavior and properties. However, this is difficult to achieve using conventional genetic approaches.

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