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Published on: February 24, 2023
Live-cell Imaging and Quantitative Analysis of Meiotic Divisions in Caenorhabditis elegans Males
Gunar Fabig1, Falko Löffler2, Christian Götze2
1Experimental Center, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany.
Abstract:
Live-imaging of meiotic cell division has been performed in extracted spermatocytes of a number of species using phase-contrast microscopy. For the nematode Caenorhabditis elegans, removal of spermatocytes from gonads has damaging effects, as most of the extracted spermatocytes show a high variability in the timing of meiotic divisions or simply arrest during the experiment. Therefore, we developed a live-cell imaging approach for in situ filming of spermatocyte meiosis in whole immobilized C. elegans males, thus allowing an observation of male germ cells within an unperturbed environment. For this, we make use of strains with fluorescently labeled chromosomes and centrosomes. Here we describe how to immobilize male worms for live-imaging. Further, we describe the workflow for the acquisition and processing of data to obtain quantitative information about the dynamics of chromosome segregation in spermatocyte meiosis I and II. In addition, our newly developed approach allows us to re-orient filmed spindles in silico, regardless of the initial 3D orientation in the worm, and analyze spindle dynamics in living worms in a statistically robust manner. Our live-imaging approach is also applicable to C. elegans hermaphrodites and should be expandable to other fluorescently labelled nematodes or other fully transparent small model organisms.
Insights
We developed a new live-imaging method to study meiosis in Caenorhabditis elegans (C. elegans) spermatocytes within unperturbed worms. This approach enables robust analysis of chromosome segregation dynamics during male germ cell division.
Area of Science:
- Cell Biology
- Developmental Biology
- Genetics
Background:
- Live-imaging of meiosis in Caenorhabditis elegans (C. elegans) spermatocytes is challenging due to cell damage upon extraction.
- Existing methods show high variability or arrest of meiotic divisions in extracted C. elegans spermatocytes.
Purpose of the Study:
- To develop a novel in situ live-cell imaging approach for observing spermatocyte meiosis in whole, immobilized C. elegans males.
- To enable quantitative analysis of chromosome segregation dynamics in an unperturbed environment.
Main Methods:
- Immobilization of male C. elegans worms for live-imaging.
- Utilizing fluorescently labeled chromosomes and centrosomes in C. elegans strains.
- Developing a workflow for data acquisition, processing, and in silico re-orientation of spindles.
Main Results:
- Successfully filmed spermatocyte meiosis I and II in situ within immobilized male C. elegans.
- Enabled statistically robust analysis of chromosome segregation and spindle dynamics.
- Demonstrated the applicability to C. elegans hermaphrodites and potential for other model organisms.
Conclusions:
- The developed in situ live-imaging approach overcomes limitations of previous methods for studying C. elegans male meiosis.
- This technique provides a powerful tool for quantitative analysis of meiotic dynamics in a native cellular environment.
- The method is adaptable for broader applications in C. elegans research and other transparent model organisms.
