Zebrafish Neuroblastoma Drug Treatment

Zebrafish experiments were approved by the UCD Animal Research Ethics Committee and the Healthcare Products Regulatory Authority (AE18982/P038). All methods were performed in accordance with the relevant guidelines and regulations. The transgenic Tg[dβh:MYCN-EGFP] zebrafish line was a generous gift from Prof. A. Thomas Look (Dana-Farber Cancer Institute, Harvard Medical School)^{33 (link)}. Adult Tg[dβh:MYCN-EGFP] zebrafish (Danio rerio) were maintained on a 14 h light/10 h dark cycle at 28 °C; and monitored for fluorescent tumor masses every two weeks. Approximately 20% of fish develop the equivalent of human neuroblastoma by about 8 months of age^{33 (link)}. Drug treatments of zebrafish were performed as described earlier^{40 (link)}. Zebrafish developing tumors (age: 21 months; n = 3) were treated with YM155 and lapatinib every 72 hours for a total of seven days. Fish water was changed every 72 hours, and drugs were dissolved in the fish water. Zebrafish were anesthetized by 0.016% Tricaine prior to microscopic analysis. An Olympus SZX10 fluorescent stereo microscope equipped with an Olympus DP71 camera was used to capture images. The fluorescent area posterior to the gills, which corresponds to the region of neuroblastoma growth^{33 (link)}, was analysed using the ImageJ software (v1.44p).