Induction of Spawning in the Starlet Sea Anemone Nematostella vectensis, In Vitro Fertilization of Gametes, and Dejellying of Zygotes

Abstract

INTRODUCTIONThis protocol describes a reliable method for inducing spawning in the starlet sea anemone Nematostella vectensis under laboratory conditions. Spawning occurs at ~13±3 h after the onset of the inductive cues, which are a particular combination of light and temperature. This method has been used successfully on a daily basis with rotating cultures, yielding thousands of embryos every day. Each given female or male can be induced once per week. For long-term health, an induction once every 10-14 d is less exhausting to the animals. The protocol also describes the dejellying of the zygotes for further manipulation of the embryos.