Long-term zebrafish survival assay. (A) Experimental design of the long-term survival assay. Cohorts of sapje or sapje-like offspring were screened as affected or unaffected on 4 dpf, at which time drug treatment was initiated and continued through 30 dpf. The water was changed and surviving fish were counted every other day. (B–G) Treatment with 33 μM serotonin, 66 μM 5-HTP, 16.5 μM tryptophan, 33 μM melatonin, 8.25 μM cisapride or 33 μM moclobemine did not significantly improve the survival of affected fish. 8.25 μM cisapride was toxic to both affected and unaffected fish beginning on 14 dpf. For each condition, 30–40 fish were tested in three replicate experiments. Data represent means±s.e.m. †P<0.05 affected versus respective unaffected, *P<0.05 drug-treated versus respective control by two-way ANOVA and Bonferroni post-hoc test. AF, affected; UA, unaffected. (H) Affected fish treated with 2.5 μg/ml aminophylline significantly increased survival versus affected controls. †P<0.05 affected versus respective unaffected, *P-values are for the closed blue circles and indicate significant difference between drug-treated AF versus control AF by two-way ANOVA and Bonferroni post-hoc test.
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