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Labeling and Imaging Cells in the Zebrafish Hindbrain
Published on: July 26, 2010
Imaging intercellular calcium waves during late epiboly in intact zebrafish embryos
Sarah E Webb1, Andrew L Miller
1Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, SAR, PRC.
Zygote (Cambridge, England)
|June 28, 2003
Summary
Rhythmic calcium (Ca2+) waves in zebrafish embryos occur even with an intact chorion, validating their role in embryonic development. These waves are not an artifact of dechorionation and are crucial for gastrulation patterning.
Area of Science:
- Developmental Biology
- Cellular Biology
- Biophysics
Background:
- Previously reported rhythmic intercellular Ca2+ waves during zebrafish gastrulation using aequorin imaging.
- Concerns were raised that dechorionation might cause these Ca2+ waves, questioning their biological relevance.
Purpose of the Study:
- To investigate if rhythmic intercellular Ca2+ waves occur in intact zebrafish embryos.
- To confirm the presence and characteristics of these waves in unmanipulated embryos.
Main Methods:
- Aequorin imaging in zebrafish embryos with intact chorions.
- Ca2+-green dextran loading at the single-cell stage followed by scanning confocal microscopy.
- Comparison of wave properties between dechorionated and intact embryos.
Main Results:
- Rhythmic intercellular Ca2+ waves were observed in zebrafish embryos with intact chorions.
- Wave characteristics (onset, timing, velocity, pathway) were consistent between intact and dechorionated embryos.
- Confocal microscopy confirmed circumferential Ca2+ waves in deep cells at the blastoderm margin.
Conclusions:
- The rhythmic Ca2+ waves are a genuine biological phenomenon in zebrafish development, not an artifact of dechorionation.
- These pan-embryonic Ca2+ waves likely play a fundamental role in embryonic patterning during gastrulation.
