Muscular contractions in the zebrafish embryo are necessary to reveal thiuram-induced notochord distortions

Teraoka, H., Urakawa, S., Nanba, S., Nagai, Y., Dong, W., Imagawa, T., Tanguay, R.L., Svoboda, K., Handley-Goldstone, H.M., Stegeman, J.J., and Hiraga, T.
Toxicology and applied pharmacology   212(1): 24-34 (Journal)
Registered Authors
Dong, Wu, Goldstone, Heather M. H., Stegeman, John J., Svoboda, Kurt, Tanguay, Robyn L., Teraoka, Hiroki
Agricultural chemical; Pesticide; Thiram; TMTD; Vertebral malformation
MeSH Terms
  • Animals
  • Chelating Agents/pharmacology
  • Collagen/metabolism
  • Dose-Response Relationship, Drug
  • Embryo, Nonmammalian/drug effects
  • Embryo, Nonmammalian/physiology*
  • Fungicides, Industrial/toxicity*
  • In Situ Hybridization
  • Laminin/metabolism
  • Muscle Contraction/drug effects
  • Muscle Contraction/physiology
  • Muscle, Skeletal/abnormalities
  • Muscle, Skeletal/physiology*
  • Notochord/abnormalities*
  • Thiocarbamates/toxicity
  • Thiram/toxicity*
  • Zebrafish/physiology*
16051294 Full text @ Tox. App. Pharmacol.
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 mug/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 EC(50) 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.
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