Maturation-associated changes in internal distribution and intra-ovarian microdistribution of tetrodotoxin in the pufferfish Takifugu pardalis

To clarify the maturation-associated accumulation profile of tetrodotoxin (TTX) in the female pufferfish Takifugu pardalis, specimens collected from Omura Bay, Nagasaki Prefecture, Japan, from November through December, 2014, were separated based on the histology of ovarian sections into four groups: yolk vesicle stage (Yv), yolk globule stage (Yg)-1, Yg-2, and Yg-3. TTX quantification in each tissue in each group by liquid chromatography–mass spectrometry revealed that the TTX concentration [mouse unit (MU)/g] in the ovary increased from Yv to Yg-1, and the TTX amount (MU/individual) in the ovary increased from Yg-1 through Yg-3. In Yv, TTX in the skin accounted for 86% of the total TTX in the body, whereas from Yg-1 through Yg-3, ovarian TTX accounted for 53–76% of the total. In female specimens collected from the same bay from February to May 2015, the TTX amount in the ovary markedly decreased after spawning, and the TTX amount in the liver increased. Immunohistochemical observation of the ovarian sections indicated that TTX localized mainly in the nucleus of immature oocytes, but then distributes to the yolk vesicles and yolk globules as maturation advances, and is partly transferred to the egg membrane as spawning approaches.