ABSTRACT
To investigate seasonal changes in the whole body content and tissue distribution of tetrodotoxin (TTX) in the pufferfish Takifugu niphobles, wild individuals were collected from the coastal waters around Miura Peninsula from November 2010 to May 2012, and their tissues were subjected to LC-MS/MS analysis. Fish that were sexually mature were classified as being in the maturation period (April), the spawning period (May-July) or the "ordinary period" (i.e., other months). In both sexes, gonad somatic index rapidly increased during the maturation period and then decreased during the spawning period. Whole body TTX content was significantly higher during the maturation/spawning period than in the ordinary period. Through all seasons, TTX was localized in the skin or ovary in females and in the skin or liver in males: the difference in TTX localization between females and males was particularly evident during the spawning period.
Subject(s)
Seasons , Takifugu/metabolism , Tetrodotoxin/metabolism , Animals , Chromatography, Liquid , Female , Liver/metabolism , Male , Ovary/metabolism , Sexual Behavior, Animal , Sexual Maturation , Skin/metabolism , Tandem Mass Spectrometry , Tetrodotoxin/isolation & purificationABSTRACT
Marine pufferfish contain tetrodotoxin (TTX), an extremely potent neurotoxin. All species of the genus Takifugu accumulate TTX in the liver and ovaries, although the tissue(s) in which it is localized can differ among species. TTX is the major defense strategy the pufferfish appears to use against predators. TTX is also used as a male-attracting pheromone during spawning. Here we demonstrate an additional (and unexpected) use of maternal TTX in the early larval stages of the Takifugu pufferfish. Predation experiments demonstrated that juveniles of all the species of fish used as predators ingested pufferfish larvae, but spat them out promptly. Liquid Chromatography-Tandem Mass Spectrometry (LC-MSMS) analysis revealed that the pufferfish larvae contain a small quantity of TTX, which is not enough to be lethal to the predators. Immunohistochemical analysis with anti-TTX monoclonal antibody revealed that the TTX is primarily localized in the body surface of the larvae as a layer of protection. Our study showed the female parent of the Takifugu pufferfish vertically transfers TTX to the larvae through its accumulation in the ovaries, and subsequent localization on the body surface of the larvae.
