Fig 1.
Assessment of embryonic tissue ethanol concentration compared to waterbath ethanol concentration following acute 24-hour ethanol exposure of zebrafish embryos.
A Alcohol was assayed in whole embryos developmentally exposed to an ethanol waterbath concentration of 100mM and 20mM for a period of 24 hours, using 1-24hpf, 24-48hpf and 48-72hpf periods. B Tissue ethanol content after exposure to 100mM ethanol during the whole period 1-9dpf. Values are means ± SE, 3 samples collected on different days were used for each time period, 25 animals per sample. Values for tissue ethanol in the absence of added ambient ethanol were less than 1% of those illustrated, and not significantly different from zero.
Fig 2.
Effects of ethanol exposure from 1-9dpf on larval development A.
size at 9dpf measured using eLaborant software. B dry weight at 9dpf. C body weight at 6 months. There were no differences between groups. Values are means ± SE, 3 samples were used per group, with 20 (A) or 25 (B) animals per sample.
Fig 3.
Stress-reactivity in 9dpf larvae measured by whole-body cortisol concentration.
Experimental animals were developmentally treated with ethanol from 1dpf-9dpf with 20mM and 50mM ethanol concentrations. At 9dpf animals were either flash frozen immediately or air exposed then frozen. Air stressed animals showed decreased tissue cortisol concentrations with increasing ethanol concentration exposure during development. Means ± SE, 6 batches of animals per group, *P< 0.05, **P<0.01, by ANOVA and “t” test.
Fig 4.
Stress-reactivity measured by whole-body cortisol concentration of zebrafish 6-month-old adults.
Animals were either flash frozen or stressed, either by air exposure at a specific time or acute exposure to ethanol (1%), followed by freezing 6 minutes later. Zebrafish adults showed increased cortisol concentration when air exposed for 30s, but not after 60s exposure. Means ± SE, 3 batches (on different days) of 3 samples of 10 animals were used. P<0.01, ANOVA <0.01).
Fig 5.
Stress-reactivity measured by whole-body cortisol concentration of zebrafish 6-month-old adults.
Treated animals were developmentally exposed to ethanol from 1dpf-9dpf with 20mM or 50mM ethanol concentrations. At 6 months animals were either flash frozen immediately or air exposed and frozen 6 minutes later. Zebrafish adults showed decreased cortisol content with increasing ethanol concentration exposure during development. Means ± SE, 9 batches of animals per group. *P<0.05 **P<0.01 “t” test and ANOVA.