Neural Crest Methodologies in Zebrafish and Medaka.
Kleio Petratou1, Karen Camargo-Sosa2, Ruqaiya Al Jabri2
1Institute for Cardiovascular Organogenesis and Regeneration, Faculty of Medicine, Westfälische Wilhelms-Universität Münster, Münster, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|April 13, 2019
Summary
This study details methods for analyzing neural crest cell development in fish embryos. Protocols for whole-mount in situ hybridization and RNAscope enable precise identification of these migratory cells.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Genetics
Background:
- Neural crest cells are multipotent and migratory, crucial for homeostasis and regeneration.
- Fish embryos offer optical transparency for studying neural crest development.
- Cell dispersion necessitates robust identification methods using marker expression.
Purpose of the Study:
- To describe key protocols for analyzing neural crest marker expression in zebrafish and medaka.
- To provide methods for high-resolution analysis of neural crest development in fish embryos.
Main Methods:
- Whole-mount in situ hybridization for mRNA detection using chromogenic substrates.
- RNAscope for multiplexed fluorescent detection of mRNA.
- Immunofluorescent detection of antigens.
Main Results:
- Established protocols for analyzing neural crest marker expression in zebrafish and medaka.
- Demonstrated the utility of RNAscope for multiplexed fluorescent detection.
- Validated immunofluorescent detection of antigens.
Conclusions:
- The described protocols facilitate detailed analysis of neural crest cell development in fish.
- These methods aid in understanding the roles of neural crest cells in homeostasis and regeneration.
- Advanced techniques like RNAscope enhance the study of dispersive cell populations.
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