Dithiocarbamates form a large group of chemicals that have numerous uses in agriculture and medicine. It has been reported that dithiocarbamates, including thiuram (tetramethylthiuram disulfide), cause wavy distortions of the notochord in zebrafish and other fish embryos. In the present study, we investigated the mechanism underlying the toxicity of thiuram in zebrafish embryos. When embryos were exposed to thiuram (2-1000 nM: 0.48-240 microg/L) from 3 h post fertilization (hpf) (30% epiboly) until 24 hpf (Prim-5), all embryos develop wavy notochords, disorganized somites, and have shortened yolk sac extensions. The thiuram response was specific and did not cause growth retardation or mortality at 24 hpf. The thiuram-dependent responses showed the same concentration dependence with a waterborne EC50 values of approximately 7 nM. Morphometric measurements revealed that thiuram does not affect the rate of notochord lengthening. However, the rate of overall body lengthening was significantly reduced in thiuram-exposed animals. Other dithiocarbamates, such as ziram, caused similar malformations to thiuram. While expression of genes involved in somitogenesis was not affected, the levels of notochord-specific transcripts were altered after the onset of malformations. Distortion of the notochord started precisely at 18 hpf, which is concomitant with onset of spontaneous rhythmic trunk contractions. Abolishment of spontaneous contractions using tricaine, alpha-bungarotoxin, and a paralytic mutant sofa potato, resulted in normal notochord morphology in the presence of thiuram. These results indicate that muscle activity is necessary to reveal the underlying functional deficit and suggest that the developmental target of dithiocarbamates impairs trunk plasticity through an unknown mechanism.

ABSTRACT. Organotins are among the most common marine pollutants in the world, as they are widely used as antifouling paint on ships and fishing nets. It has been reported that organotin preferentially accumulates in the central nervous system, especially in the retinal neurons of marine organisms including fish. In this study, we investigated the effects of waterborne tributyltin (TBT) on early-stage developing zebrafish (Danio rerio). Below the lethal concentrations, TBT specifically increased the number of apoptotic cells in the retina as well as some cells near trigeminal neurons, detected by terminal transferase-mediated nick-end-labeling staining. Apoptosis peaked at 60 hpf and decreased by 72 hpf, which was associated with macrophage accumulation. Furthermore, the effect of TBT was markedly inhibited by antioxidants, ascorbic acid or trolox. These results suggest that TBT preferentially induces apoptosis in the retinal neuron of developing zebrafish. Oxidative stress may be involved in this toxicological response.