title: Muscular contractions in the zebrafish embryo are necessary to reveal thiuram-induced notochord distortions.
abstract: 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.
authors.affiliation: Department of Toxicology, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, Japan. hteraoka@rakuno.ac.jp
authors.forename: Hiroki
authors.initials: H
authors.lastname: Teraoka
authors.forename: Satsuki
authors.initials: S
authors.lastname: Urakawa
authors.forename: Satomi
authors.initials: S
authors.lastname: Nanba
authors.forename: Yuhki
authors.initials: Y
authors.lastname: Nagai
authors.forename: Wu
authors.initials: W
authors.lastname: Dong
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Imagawa
authors.forename: Robert L
authors.initials: RL
authors.lastname: Tanguay
authors.forename: Kurt
authors.initials: K
authors.lastname: Svoboda
authors.forename: Heather M
authors.initials: HM
authors.lastname: Handley-Goldstone
authors.forename: John J
authors.initials: JJ
authors.lastname: Stegeman
authors.forename: Takeo
authors.initials: T
authors.lastname: Hiraga
mesh_terms: D000818:Animals; D002614:Chelating Agents / Q000494:pharmacology; D003094:Collagen / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000502:physiology*; D005659:Fungicides, Industrial / Q000633:toxicity*; D017403:In Situ Hybridization; D007797:Laminin / Q000378:metabolism; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology; D018482:Muscle, Skeletal / Q000002:abnormalities / Q000502:physiology*; D009672:Notochord / Q000002:abnormalities*; D013859:Thiocarbamates / Q000633:toxicity; D013893:Thiram / Q000633:toxicity*; D015027:Zebrafish / Q000502:physiology*
chemical_list: D002614:Chelating Agents; D005659:Fungicides, Industrial; D007797:Laminin; D013859:Thiocarbamates; D013893:Thiram; D003094:Collagen