NONYLPHENOL INDUCES EXPRESSION OF THE T-BOX6 GENE IN ZEBRAFISH EMBRYOS: RESULTS(1)

The development of zebrafish embryos after incubation with various isomers of NP is demonstrated in Figure 1. Normal development after 48 hr of incubation results in a long extended tail without any bending or tip broadening (Figure 1H). The malformations in development of the tail produced by treatment with different isomers of NP are shown in Figure 1A-G. In some cases the malformation were accompanied by necrosis of the tail tip (results not shown). All NP isomers produced spinal malformations: Zebrafish embryos treated with p22-NP showed short tails which appeared to be incomplete developed (Figure 1A). Exposure of zebrafish embryos to p252-NP, p262-NP, p33-NP and /-NP results in bended tails. These tails showed a sharp kink in the middle of the tail length combined with a turn in the tail orientation (Figure 1B, C, D and G). Since the concentration of p22-NP and p33-NP required to induce the malformation described was 50^mol/l, calculation of EC50 was not possible. p353-NP produced a tail tip broadening (Figure 1E) which was described previously 23. At a concentration of 16 ^mol/l 50% of the embryos showed that deformation (Table1).

Slight swelling of the tail tip was observed after exposure of embryos to p363-NP (Figure 1F). The phenotype of tail tip broadening is unique for p353-NP and was never seen with other isomers of NP. After 1 hr of treatment of 3-4 hr old embryos no marked differences in phenotypes could be detected. The first time when tail tip shape was visible was around 24 hr after fertilization – independent of the treatment. No general alterations in the tail were observed from then until 48 hr after fertilization, when the tail tip was visible best. This time frame was selected because in preliminary experiments one could demonstrate that a change in expression of the selected genes is observable after 1 hr exposure, demonstrating that gene activation of transcription factors is rapid.

Fig1Nonylphenol Induces Expression_decrypted

Figure 1: 48h old zebrafish embryos after exposition with different isomers of nonylphenols (A-F), technical mixture of nonylphenol (G) and a control embryo without any treatment (H). The arrows indicate the tail malformations: White: bended tail; black: broadened tail tip; grey: very short tail.

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