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Related Experiment Video

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Basic Caenorhabditis elegans Methods: Synchronization and Observation
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Using fast-acting temperature-sensitive mutants to study cell division in Caenorhabditis elegans.

T Davies1, S Sundaramoorthy1, S N Jordan1

  • 1Columbia University, New York, NY, United States.

Methods in Cell Biology
|January 10, 2017
PubMed
Summary

Fast-acting temperature-sensitive (ts) mutations enable precise study of cell division (cytokinesis) in C. elegans. These mutants allow rapid, reversible protein inactivation by temperature shifts, offering functional tunability for transient cellular processes.

Keywords:
C. elegansCell divisionCherryTempCytokinesisPermissive temperatureRapid thermal upshiftRestrictive temperatureTemperature control deviceTemperature-sensitive mutantTherminatorTs mutant

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Area of Science:

  • Cell Biology
  • Developmental Biology
  • Genetics

Background:

  • Fast-acting temperature-sensitive (ts) mutations are crucial for studying dynamic cellular processes like cytokinesis.
  • These mutants function normally at permissive temperatures but exhibit complete loss-of-function at restrictive temperatures.
  • The ability to rapidly and reversibly inactivate proteins via temperature shifts provides significant experimental control.

Purpose of the Study:

  • To describe techniques for utilizing fast-acting ts mutants to investigate cytokinesis in Caenorhabditis elegans.
  • To present methods for precise temporal control over protein function during cell division.
  • To offer protocols for embryo preparation and imaging in C. elegans.

Main Methods:

  • Utilizing fast-acting ts cytokinesis-defective mutants in C. elegans.
  • Employing passive heat transfer and advanced fluidic-based thermal control systems for temperature shifts.
  • Standard dissection, mounting, and imaging of early worm embryos.

Main Results:

  • Demonstration of functional tunability and rapid protein inactivation using ts mutants.
  • Successful application of thermal control systems to induce cytokinesis failure.
  • Detailed protocols provided for experimental procedures.

Conclusions:

  • Fast-acting ts mutants are powerful tools for dissecting the temporal regulation of cytokinesis.
  • Temperature-shift protocols allow precise manipulation of protein function during cell division.
  • The described methods facilitate detailed analysis of cytokinesis in C. elegans embryos.