Dithiocarbamates induce craniofacial abnormalities and downregulate sox9a during zebrafish development.
Dithiocarbamates (DTCs) have a wide variety of applications in diverse fields ranging from agriculture to medicine. DTCs are teratogenic to vertebrates but the mechanisms by which they exert these effects are poorly understood. Here, we show that low nanomolar exposure to three DTCs, tetraethylthiuram (thiram), tetramethylthiuram (disulfiram), and sodium metam (metam), leads to craniofacial abnormalities in developing zebrafish embryos that are reminiscent of DTC-induced abnormalities found in higher vertebrates. In order to better understand the molecular events underlying DTC teratogenesis, we exposed embryonic zebrafish (PAC2) cells to thiram and disulfiram and measured changes in gene expression with microarrays. We found differential expression of 166 genes that were specific for exposure to DTCs and identified a network of genes related to connective tissue development and function. Additionally, we found eight downregulated genes related to transforming growth factor beta-1 (TGF-beta1) signaling, including an essential transcription factor for zebrafish craniofacial development, SRY-box-containing gene 9a (sox9a). Finally, we show that sox9a expression is perturbed in the ceratobranchial arches of DTC-exposed zebrafish, suggesting that this is an important event in the development of DTC-induced craniofacial abnormalities. Together, we provide evidence for a novel teratogenic endpoint and a molecular basis for a better understanding of DTC-induced teratogenesis in vertebrates.
Institute for Environmental Studies (IVM), VU University, Amsterdam, The Netherlands.
Antonius Leonardus
AL
van Boxtel
Bart
B
Pieterse
Peter
P
Cenijn
Jorke Harmen
JH
Kamstra
Abraham
A
Brouwer
Wessel
W
van Wieringen
Jacob
J
de Boer
Juliette
J
Legler
D000818:Animals; D001483:Base Sequence; D019465:Craniofacial Abnormalities / Q000139:chemically induced*; D017931:DNA Primers; D015536:Down-Regulation / Q000187:drug effects*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D017403:In Situ Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics*; D013723:Teratogens / Q000633:toxicity*; D013859:Thiocarbamates / Q000633:toxicity*; D015027:Zebrafish / Q000196:embryology*; D029961:Zebrafish Proteins / Q000235:genetics*
D017931:DNA Primers; D055758:SOX9 Transcription Factor; C504918:Sox9a protein, zebrafish; D013723:Teratogens; D013859:Thiocarbamates; D029961:Zebrafish Proteins