Capturing Cytoskeleton-Based Agitation of the Mouse Oocyte Nucleus Across Spatial Scales

Ga&#235;lle Letort; Philippe Mailly; Adel Al Jord; Maria Almonacid

doi:10.3791/65976

Capturing Cytoskeleton-Based Agitation of the Mouse Oocyte Nucleus Across Spatial Scales

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Biology

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JoVE Journal Biology

Capturing Cytoskeleton-Based Agitation of the Mouse Oocyte Nucleus Across Spatial Scales

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05:43 min

January 12, 2024

DOI: 10.3791/65976-v

Gaëlle Letort^1,2, Philippe Mailly¹, Adel Al Jord^1,3, Maria Almonacid¹

¹Center for Interdisciplinary Research in Biology (CIRB),Collège de France, CNRS, INSERM, Université PSL, ²Department of Developmental and Stem Cell Biology,Institut Pasteur, Université de Paris Cité, CNRS, ³Centre for Genomic Regulation (CRG),The Barcelona Institute of Science and Technology

Overview

This study explores the mechanical forces transmitted by the cytoskeleton to the nucleus in mouse oocytes, a key aspect of understanding female gamete biology. By analyzing cytoskeletal impacts using advanced imaging techniques, the research highlights the dynamics of nuclear shape and internal organelles in oocytes.

Key Study Components

Research Area

Cellular mechanics
Female gamete biology
Nuclear architecture

Background

Importance of understanding gamete quantity and quality
The role of the cytoskeleton in nuclear mechanics
Use of mouse oocyte as a model system

Methods Used

Imaging and image analysis techniques
Mouse oocyte model
Non-invasive methods for assessing nuclear mechanics

Main Results

Control oocytes showed significant fluctuations in nuclear shape, indicative of cytoskeletal activity.
Disruption of cytoskeletal forces led to increased stability in nuclear shape.
Findings suggest a link between cytoskeletal dynamics and RNA processing within the nucleus.

Conclusions

The study demonstrates how cytoskeletal forces influence nuclear shape and RNA dynamics.
These insights can inform reproductive biology and cellular mechanics research.

Frequently Asked Questions

What is the primary focus of this research?

The research focuses on understanding how cytoskeletal forces affect nuclear shape and internal organelles in mouse oocytes.

Why are mouse oocytes used as a model system?

Mouse oocytes provide a key model for studying female gamete mechanisms essential for reproduction.

What methods were employed in this study?

The study utilized advanced imaging techniques along with mechanical assessments of the nucleus.

What were the main findings regarding nuclear fluctuations?

Control oocytes exhibited significant peripheral fluctuations in their nuclei compared to those with disrupted cytoskeletal forces.

How does this research contribute to biology?

It provides insights into the mechanical properties of cells and their impact on reproductive biology.

What implications does this research have for understanding gametes?

The findings may help improve knowledge on female fertility and gamete development processes.

This protocol provides an experimental framework to document the physical impact of the cytoskeleton on nuclear shape and the internal membrane-less organelles in the mouse oocyte system. The framework can be adapted for use in other cell types and contexts.

The lab aims to better understand the mechanisms involved in the acquisition of female gamete quantity, which is essential for the reproduction of sexual species using the mouse oocyte model system. Recently we identified a novel mechanical transduction process that promotes agitation of the nucleus and its content leading to the fine regulation of maternal RNA in mouse oocytes. The imaging and image analysis pipeline presented in this protocol allows us to highlight the transmission of cytoskeletal forces as a continuum across scales from the cytoplasm to the nucleus and its components, including nuclear RNA processing bodies in mouse oocytes.

This protocol provides simple tools that allow us to address first transmission across multiple cellular scales in a single pipeline for the first time. Complemented with biophysical modeling, this protocol constitutes a non-invasive technique to address changes in nuclear mechanics and the dissipation of energy across cellular compartments. Begin by arranging all the materials required for the experiment on the working platform.

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