Long-term behavioral impairment following acute embryonic ethanol exposure in zebrafish

• Zebrafish were exposed to 1 or 3% ethanol 8–10 or 24–27 hpf to evaluate long-term behavioral effects.
• Ethanol exposure during the 24–27 hpf window caused locomotor hyperactivity during adolescence.
• Behavioral effects were detected at doses that did not cause physical malformation.
• Behavioral phenotypes of embryonic ethanol exposure are determined by window of exposure.

BackgroundDevelopmental exposure to ethanol has long been known to cause persisting neurobehavioral impairment. However, the neural and behavioral mechanisms underlying these deficits and the importance of exposure timing are not well-characterized. Given the importance of timing and sequence in neurodevelopment it would be expected that alcohol intoxication at different developmental periods would result in distinct neurobehavioral consequences.MethodsZebrafish embryos were exposed to ethanol (0%, 1%, 3%) at either 8–10 or 24–27 h post-fertilization (hpf) then reared to adolescence and evaluated on several behavioral endpoints. Habituation to a repeated environmental stimulus and overall sensorimotor function were assessed using a tap startle test; measurements of anxiety and exploration behavior were made following introduction to a novel tank; and spatial discrimination learning was assessed using aversive control in a three-chambered apparatus. Overt signs of dysmorphogenesis were also scored (i.e. craniofacial malformations, including eye diameter and midbrain-hindbrain boundary morphology).ResultsEthanol treated fish were more active both at baseline and following a tap stimulus compared to the control fish and were hyperactive when placed in a novel tank. These effects were more prominent following exposure at 24–27 hpf than with the earlier exposure window, for both dose groups. Increases in physical malformation were only present in the 3% ethanol group; all malformed fish were excluded from behavioral testing.DiscussionThese results suggest specific domains of behavior are affected following ethanol exposure, with some but not all of the tests revealing significant impairment. The behavioral phenotypes following distinct exposure windows described here can be used to help link cellular and molecular mechanisms of developmental ethanol exposure to functional neurobehavioral effects.