Spawning induced by cubifrin in the Japanese common sea cucumber Apostichopus japonicus

The neuropeptide cubifrin-I and its derivative cubifrin-L have recently been shown to be potent substances that induce in vitro oocyte maturation and spawning in the Japanese common sea cucumber Apostichopus japonicus. We have examined the reproductive behavior of A. japonicus in vivo following the injection of cubifrin-L into the body cavity with the aim of determining the practical value of using this peptide to induce spawning in the hatchery setting. The responsiveness of ovarian fragments with oocytes >155 μm in diameter to cubifrin-L in vitro was well correlated with the spawning success induced in vivo by a cubifrin-L injection. Mature sea cucumbers injected with cubifrin-L displayed sequential reproductive behaviors, which comprised climbing up the side wall of the tank toward the water surface, waving of the head, and shedding of gametes. Gamete shedding started about 60 and 80 min after the injection of cubifrin-L in males and females, respectively, and was completed almost simultaneously in the two sexes about 2 h post-injection. Repeated injections of cubifrin-L at intervals of about 10 days successfully induced multiple spawnings in males and females. These results demonstrate that cubifrin-L can be used as an effective inducer of spawning in Japanese sea cucumber cultivation.