Long-term behavioral alterations following embryonic alcohol exposure in three zebrafish populations.

Fetal alcohol exposure may lead to a condition known as fetal alcohol spectrum disorder (FASD), which comprises a set of consequences, including cognitive and behavioral impairments. Although zebrafish has been applied as a reliable model for studying FASD, there is no approach to the disorder's ontogeny and population differences. Here, we evaluated the behavioral outcomes of AB, Outbred (OB), and Tübingen (TU) zebrafish populations embryonically exposed to alcohol throughout the development to the adult stage. We exposed 24hpf eggs to 0 %, 0.5 %, or 1.0 % alcohol for 2 h. Fish were let grow and locomotor and anxiety-like behaviors were tested in a novel tank at larval - 6dpf, juvenile - 45dpf, and adult- 90dpf stages. At 6dpf, both AB and OB treated with 1.0 % alcohol showed hyperactivity, while 0.5 % and 1.0 % TU fish exhibited hypolocomotion. At 45dpf, AB and TU fish maintained the larval pattern of locomotion. At the adult stage - 90dpf, both AB and TU populations showed increased locomotor activity and anxiogenic responses, while the OB population did not show altered behavior. Our results show for the first time that zebrafish populations exhibit behavioral differences in response to embryonic alcohol exposure and that it varies along animals' ontogeny. AB fish showed the most consistent behavioral pattern through developmental stages, TU fish showed behavioral changes only in adulthood, and OB population showed high interindividual variability. These data reinforce that different populations of zebrafish are better adapted to translational studies, offering reliable results in contrast to domesticated OB populations obtained from farms, which exhibit more variable genomes.

Variation in nest building, aggression, learning, and steroid hormone levels in Betta splendens.

Individuals within a population present behavioral responses that vary according to intrinsic and extrinsic factors such as ontogenetic phase, nutritional status, reproductive stage, and previous experiences. These differences can be explained by endogenous changes, such as hormone release, that can modulate reproductive behaviors, stress response, and cognitive processes. In order to investigate the relationship between behavior and hormonal levels in the fighting fish Betta splendens, the present study characterized nest building, aggressive behavior, learning of a task, and levels of cortisol and 11-ketotestosterone (11-KT) in 86 male Fighting fish. At the beginning of the experiment (days 1-4), fish were characterized as nest builders, intermediate builders, or non-builders. They were then sequentially tested for aggression (days 7-8), learning performance (days 11-21), and circulating hormone levels (day 23). Nest builders showed the lowest hormonal levels at the end of the experiment and low aggressiveness; Intermediate builders presented low cortisol, but high KT levels and best learned the task; Non-builders were the most aggressive animals with higher cortisol levels (at day 23). Our data suggest that in B. splendens, aggressive behavior and learning performance are related to the relative investment in reproduction and variation in circulating levels of corticosteroids and androgens.

Behavioral genetics of alcohol's effects in three zebrafish (Danio rerio) populations.

Alcohol abuse is one of the most dangerous and serious problems for patients and society. Interpopulation studies are important in understanding how genetic background contributes to the effects of alcohol. In this study, we applied a chronic alcohol exposure protocol in three zebrafish populations (Danio rerio; both sexes; AB, TU, and outbred fish - OB). We analyzed the behavioral responses and mRNA expression involved in neurotransmitter metabolism - th1, tph1, ache, ada1, gaba1, gad1b, and bdnf. Locomotion patterns were similar between populations (increased speed after acute alcohol and unaltered locomotion after chronic and withdrawal treatments). All populations exhibited increased expression of genes associated with locomotion (th1, gad1b, and gaba1) after acute alcohol exposure. Anxiety-like responses increased in AB and TU fish during withdrawal and decreased in AB fish after acute alcohol exposure. Genes related to anxiety-like behavior (tph1 and ada1) were overexpressed in AB and TU fish after acute and withdrawal treatments, while OB fish exhibited unaltered responses. Bdnf levels decreased during withdrawal in AB and OB fish, while TU showed upregulated levels in both chronic and withdrawal treatments. Our results suggest that zebrafish populations respond differently to alcohol exposure, which may contribute to understanding the mechanisms underlying alcohol use and dependence. Moreover, we found that a more diverse genetic background (OB) was related to higher variability in behavioral and mRNA expression, demonstrating that inbred populations (AB and TU) may be useful tools in identifying alcohol use and abuse mechanisms.

Embryonic Exposure to Ethanol Increases Anxiety-Like Behavior in Fry Zebrafish.

AIMS: Fetal alcohol spectrum disorder (FASD) is an umbrella term to describe the effects of ethanol (Eth) exposure during embryonic development, including several conditions from malformation to cognitive deficits. Zebrafish (Danio rerio) are a translational model popularly applied in brain disorders and drug screening studies due to its genetic and physiology homology to humans added to its transparent eggs and fast development. In this study, we investigated how early ethanol exposure affects zebrafish behavior during the initial growth phase. METHODS: Fish eggs were exposed to 0.0 (control), 0.25 and 0.5% ethanol at 24 h post-fertilization. Later, fry zebrafish (10 days old) were tested in a novel tank task and an inhibitory avoidance protocol to inquire about morphology and behavioral alterations. RESULTS: Analysis of variance showed that ethanol doses of 0.25 and 0.5% do not cause morphological malformations and did not impair associative learning but increased anxiety-like behavior responses and lower exploratory behavior when compared to the control. CONCLUSION: Our results demonstrate that one can detect behavioral abnormalities in the zebrafish induced by embryonic ethanol as early as 10 days post-fertilization and that alcohol increases anxious behavior during young development in zebrafish.