Whole-mount in situ TUNEL method revealed ectopic pattern of apoptosis in cadmium treated naupliar larvae of barnacle (Balanus amphitrite Darwin)

The effect of cadmium on stage II naupliar larvae of barnacle (Balanus amphitrite Darwin) was investigated. Barnacle larvae were exposed to 0-15 μM CdCl₂ for 24 h. Apoptotic cells were stained by the terminal deoxyribonucleotidyl transferase mediated dUTP nick end labelling (TUNEL) method. Incidence of apoptosis, as measured by numbers of animals with ectopic pattern of apoptosis as well as numbers of apoptotic cells per animal, was assessed using confocal microscopy. An increase in incidence of apoptosis was observed in the experimental animals with an increase in cadmium concentration. Mortality increased, and motility decreased, when barnacle larvae were exposed to an increasing concentration of cadmium. The relationship between the occurrence of apoptosis and swimming behaviour was investigated in larvae exposed to 10 μM CdCl₂. Significant increases in apoptosis were detected in the non-motile and dead nauplii. This study suggested that whole-mount in situ TUNEL method may be used to study increased occurrence of apoptotic cells in crustacean larvae.