Toxification of puffer and tetrodotoxin-associated proteins in liver

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The liver tissue slices of pufferfish accumulate in vitro tetrodotoxin (TTX), when incubated with minimum essential medium containing TTX. In the case of Takifugu rubripes liver slices incubated at a concentration of 25μg TTT/ml, TTX of 3.9μg/g was first detected at 2h and increased to 15.0μg/g at 48h. The TTX content accumulated was first detected at 2h and increased to 15.0μg/g at 48h. The TTX content accumulated was not decreased, even when the slices were further incubated without TTX for additional 48h. On the contrary, in the cases of the liver slices from parrot-bass Oplegnathus fasciatus, green ling Hexagrammos otakii and filefish Thamnaconus modestus incubated at a concentration of 25μg TTX/ml, TTX of 3-4μg/g was detected even at 0.5h. However, no significant change in TTX contents was recognized during the incubation for 48h. Further incubation of the filefish liver slices without TTX for additional 48h did not decrease the TTX content. It is unlikely that the liver slices of slices of filefish as well as pufferfish rapidly excrete TTX.Effect of inhibitors of transport system on in vitro accumulation of TTX in T.rubripes liver tissue slices was examined. Addition of p-aminohippuric acid and carnitine to the culture media significantly decreased in TTX content accumulated in the puffer liver tissue slices while that of tetraethylammonium did not, suggesting the involvement of an organic ion transporter in TTX accumulation in puffer liver.