Distinction of pufferfish species by gel electrophoresis

Abstract

Four species of the genus Lagocephalus pufferfish: L. gloveri (“kurosabafugu”), L. wheeleri (“shirosabafugu”), L. inermis (“kanafugu”) and L. lunaris (“dokusabafugu”) were discerned by the comparison of electrophoretic patterns of proteins and of L-Lactate dehydrogenase (LDH, EC 1.1.1.27) isozymes in skeletal muscle and vitreous humor. The proteins of skeletal muscle and vitreous humor were separated by polyacrylamide gel isoelectric focusing (PAG-IEF), and LDH isozymes were separated by agarose gel electrophoresis. The electrophoretograms of proteins of skeletal muscle and vitreous humor were significantly different from each other among the three species except for L. inermis. LDH zymograms in vitreous humor showed species specific patterns among the four species. Furthermore, a puncture of an eye ball of pufferfish enabled us to yield enough amount of sample to obtain the LDH zymograms not to give any damage to the fish. From these results, the comparison of LDH zymograms in vitreous humor is a useful method for distinction of these pufferfishes without making them unmarketable.