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Related Experiment Video

Updated: Jun 18, 2025

In Vivo Visualization of Calcium Transients during Fertilization and Early Development in C. elegans
05:11

In Vivo Visualization of Calcium Transients during Fertilization and Early Development in C. elegans

Published on: July 12, 2024

350

In Vivo Visualization of Calcium Transients during Fertilization and Early Development in C. elegans.

Michaela Foley1, Katie Toperzer1, Sara Sornat1

  • 1Florida Institute of Technology.

Journal of Visualized Experiments : Jove
|July 29, 2024
PubMed
Summary

Researchers developed CaFE, a calcium reporter in C. elegans, to visualize fertilization and early embryogenesis. This tool aids in studying oocyte-to-embryo transition and reveals new insights into calcium signaling during development.

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Last Updated: Jun 18, 2025

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Area of Science:

  • Developmental Biology
  • Cell Biology
  • Genetics

Background:

  • Calcium ions are crucial signaling molecules in the oocyte-to-embryo transition (OET) and early embryogenesis.
  • The nematode Caenorhabditis elegans offers unique advantages for OET studies due to its transparency and ordered gonad structure.

Purpose of the Study:

  • To develop a genetically encoded calcium indicator (CaFE) for visualizing fertilization in real-time within C. elegans.
  • To provide protocols for utilizing the CaFE reporter to dissect the OET and early embryogenesis.

Main Methods:

  • Modification of jGCaMP7s to create the CaFE reporter, engineered into a safe harbor locus.
  • Development of protocols for worm synchronization, RNAi knockdown, mounting for imaging, and calcium visualization.
  • Generation of additional C. elegans strains to support OET and embryogenesis research.

Main Results:

  • The CaFE reporter shows a robust signal upon fertilization with no significant impact on worm physiology or fecundity.
  • Demonstrated double ovulation with ipp-5 RNAi, where only the first oocyte was immediately fertilized.
  • Discovered single-cell calcium transients during early embryogenesis, indicating CaFE's persistence into development.

Conclusions:

  • The CaFE reporter is a valuable in vivo tool for studying C. elegans fertilization and embryogenesis.
  • CaFE facilitates research into cell-cell dynamics regulating internal fertilization and early development.
  • The CaFE reporter's simplicity makes it suitable for undergraduate research courses (CUREs).