Embryos of zebrafish (Danio rerio) have often been used in toxicity studies of environmentally relevant substances 20-22 An advantage of zebrafish as test organism in toxicological research is the transparency of the eggs. Spinal malformation observed after exposure of zebrafish embryos to p353-NP 23 is comparable to those reported form several groups working on developmental processes: Zebrafish expressing a mutant (non-functional) of the no tail (ntl) gene or a mutant of the spade tail gene (spt, tbxl 6) lack the notochord and tail 2’ 24-26 Studies with knock out mutants showed that homozygous spt— (double knock out embryos) lack trunk somites and trunk muscle later in development 2728. Ntl and spt proteins belong to the T-box6/16 protein subfamily of T-box transcription factors which contain a DNA binding and protein dimerization domain: The T-box 29 The T-box6/16 subfamily has important functions in the development of the mesoderm in all mammals investigated 30. The damages of ntl gene knock out and spt gene knock out phenotypes in zebrafish are less severe than mutations in or depletion of the homologous genes in other vertebrates indicating a functional redundancy among zebrafish T-box genes.

Understanding the molecular mechanisms involved in misdevelopment produced by NP is a prerequisite for further investigations in limiting the effects of NP release to the environment. Aim of the present study was to correlate the observed tail deformation produced by p353-NP with expression data of genes of the T-box6/16 subfamily. Zebrafish embryos were exposed to sublethal concentrations of different isomers of NP and real time PCR analysis for genes belonging to the T-box6/16 family was performed.

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