Establishment of culture and microinjection methods for false clownfish embryos without parental care
| Autor: | Yosuke Okada, Sakuto Yamanaka, Masato Kinoshita, Takeshi Furuta |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
animal structures
Microinjections Hatching ved/biology Water flow ved/biology.organism_classification_rank.species Embryo Cell Biology Biology biology.organism_classification Perciformes Cell biology medicine.anatomical_structure embryonic structures medicine Animals Yolk sac Model organism Paternal care Microinjection Zebrafish Developmental Biology |
| Zdroj: | Development, Growth & Differentiation. 63:459-466 |
| ISSN: | 1440-169X 0012-1592 |
| DOI: | 10.1111/dgd.12759 |
| Popis: | Anemonefish, including the false clownfish Amphiprion ocellaris, are attractive model organisms because of their unique features, such as sex change and brilliant color patterns in mutants. However, anemonefish are not widely used to study gene function using reverse genetic approaches owing to microinjection difficulties and subsequent rearing and hatching of embryos without parental care. A. ocellaris embryos are spawned on a hard substrate and cared for by their parents until hatching. However, the eggs need to be detached from the substrate and raised without their parents to perform successful microinjection. We established a method to culture and hatch A. ocellaris embryos without spawning substrates or parental care. We found that changing water and generating water flow are critical for culturing the embryos, and that water flow (as physical stimulation) and complete darkness in the dark period are necessary for successful hatching. We further investigated the effectiveness of microinjection into the yolk sac of fertilized eggs rather than into the cytoplasm, which makes microinjection easier. A reporter RNA injected into the yolk sac was transferred to the cytoplasm and translated, indicating that yolk sac microinjection is an efficient alternative as has been used for zebrafish. These findings highlight the potential of A. ocellaris as an experimental model organism for reverse genetics, and our methods could be applied to other anemonefish species. |
| Databáze: | OpenAIRE |
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