Ethanol effects on the developing zebrafish: neurobehavior and skeletal morphogenesis

Carvan, Michael J.; Loucks, Evyn J.; Weber, Daniel N.; Williams, Frederick

doi:10.1016/j.ntt.2004.06.016

Cited by 238 publications

(195 citation statements)

References 80 publications

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“…Recently three laboratories independently documented measurable ethanol-induced phenotypic features in zebrafish that could be utilized further for analysis of ethanol-responsive genes (Bilotta et al, 2004;Carvan et al, 2004;Loucks and Carvan, 2004;Reimers et al, 2004a). As shown here and in previous studies, the Japanese medaka is an alternative non-mammalian model, in addition to zebrafish, to evaluate ethanol toxicity.…”

Section: Discussionmentioning

confidence: 62%

“…Embryos at 10 dpf were used for cartilage staining in 0.1% Alcian blue (Sigma-Aldrich) as described for zebrafish by Carvan et al (2004) with minor modifications. Hatchlings (5-10/group) were anesthetized in MS 222 (0.1% in hatching solution: Sigma-Aldrich) and fixed at 41C for 24 hr in 4% paraformaldehyde (Sigma-Aldrich) in phosphate-buffered saline (PBS) with 0.1% Tween 20 (PBT).…”

Section: Cartilage Stainingmentioning

confidence: 99%

“…Large-scale mutagenic screens in zebrafish and medaka have identified several orthologue genes that are related closely to those involved in human genetic diseases (Naruse et al, 2004). The response of zebrafish embryos to ethanol has been studied by many investigators and zebrafish have been recognized as a useful model for the study of alcohol-induced teratogenic effects (Bilotta et al, 2004;Carvan et al, 2004;Lockwood et al, 2004;Reimers et al, 2004a). In zebrafish, ethanol causes cyclopia, affects visual functions, induces pericardial edema and otolith defects, lowers heart rate, reduces eye diameter, induces axial malformations, delays development, and produces axial blistering (Bilotta et al, 2002(Bilotta et al, , 2004Reimers et al, 2004a).…”

Section: Introductionmentioning

confidence: 99%

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Japanese medaka (Oryzias latipes): developmental model for the study of alcohol teratology

Wang

Williams

Haasch

et al. 2006

Birth Defects Research Pt B

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BACKGROUND: Animal models are necessary to investigate the mechanism of alcohol-induced birth defects. We have used Japanese medaka (Oryzias latipes) as a non-mammalian model to elucidate the molecular mechanism(s) of ethanol teratogenesis. METHODS: Medaka eggs, within 1 hr post-fertilization (hpf) were exposed to waterborne ethanol (0-1000 mM) in hatching solution for 48 hr. Embryo development was observed daily until 10 days post-fertilization (dpf). The concentration of embryonic ethanol was determined enzymatically. Cartilage and bones were stained by Alcian blue and calcein, respectively and skeletal and cardiovascular defects were assessed microscopically. Genetic gender of the embryos was determined by PCR. Levels of two isoenzymes of alcohol dehydrogenase (Adh) mRNAs were determined by semi-quantitative and real-time RT-PCR. RESULTS: The concentration of ethanol required to cause 50% mortality (LC 50 ) in 10 dpf embryos was 568 mM, however, the embryo absorbed only 15-20% of the waterborne ethanol at all ethanol concentrations. The length of the lower jaw and calcification in tail fin cartilaginous structures were reduced by ethanol exposure. Active blood circulation was exhibited at 501 hpf in embryos treated with 0-100 mM ethanol; active circulation was delayed and blood clots developed in embryos treated with 200-400 mM ethanol. The deleterious effects of ethanol were not gender-specific. Moreover, ethanol treatment was unable to alter the constitutive expression of either Adh5 or Adh8 mRNA in the medaka embryo. CONCLUSIONS: Preliminary results suggested that embryogenesis in medaka was significantly affected by ethanol exposure. Phenotypic features normally associated with ethanol exposure were similar to that observed in mammalian models of fetal alcohol syndrome. The results further indicated that medaka embryogenesis might be used as an alternative non-mammalian model for investigating specific alterations in gene expression as a means to understand the molecular mechanism(s) of ethanol-induced birth defects.

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Section: Discussionmentioning

confidence: 62%

Section: Cartilage Stainingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Japanese medaka (Oryzias latipes): developmental model for the study of alcohol teratology

Wang

Williams

Haasch

et al. 2006

Birth Defects Research Pt B

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“…These criteria become particularly demanding when it comes to modelling aspects of cognition. A number of recent studies have demonstrated the use of zebrafish as a valid subject for pharmacological modelling of behaviors and disorders seen in mammals (for review see Goldsmith, 2004), with a number of procedures using adult zebrafish having been developed for learning and memory (Williams et al, 2002;Carvan et al, 2004;Swain et al, 2004;Levin et al, 2006a). These have adapted rodent tests such as the T-maze and place preference for an aquatic environment.…”

Section: Discussionmentioning

confidence: 99%

Non-Associative Learning in Larval Zebrafish

Best¹,

Berghmans²,

Hunt³

et al. 2007

Neuropsychopharmacol

216

150

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Habituation, where a response is reduced when exposed to a continuous stimulus is one of the simplest forms of non-associative learning and has been shown in a number of organisms from sea slugs to rodents. However, very little has been reported in the zebrafish, a model that is gaining popularity for high-throughput compound screens. Furthermore, since most of the studies involving learning and memory in zebrafish have been conducted in adults, we sought to determine if zebrafish larvae could display non-associative learning and whether it could be modulated by compounds identified in previous rodent studies. We demonstrated that zebrafish larvae (7 days post fertilization) exhibit iterative reduction in a startle response to a series of acoustic stimuli. Furthermore, this reduction satisfied criteria for habituation: spontaneous recovery, more rapid reductions in startle to shorter intertrial intervals and dishabituation. We then investigated the pathways mediating this behavior using established compounds in learning and memory. Administration of rolipram (PDE4 inhibitor), donepezil (acetylcholinesterase inhibitor), and memantine (N-methyl-D-aspartic acid (NMDA) receptor antagonist) all increased the acoustic startle response and decreased habituation in the larvae, similar to previous rodent studies. Further studies demonstrated that NMDA blocked the memantine response and the effect of donepezil was blocked by mecamylamine but not atropine suggesting that the donepezil response was mediated by nicotinic rather than muscarinic receptors. Zebrafish larvae possess numerous advantages for medium to high-throughput screening; the model described herein therefore offers the potential to screen for additional compounds for further study on cognition function.

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“…These models have several advantages over mammalian models with regard to fertilization, development and transperancy of the egg chorion. The effects of ethanol on zebrafish embryos have been studied by a number of investigators (Bilota et al, 2004;Carvan et al, 2004;Lockwood et al, 2004;Reimers et al, 2004;Arenzana et al, 2006;Li et al, 2007;Tanguay and Reimers, 2008), establishing that the fish can be used as a model organism to study FASD. Compared to zebrafish, medaka are less utilized in studying ethanol-induced toxicity.…”

Section: Introductionmentioning

confidence: 99%

Disruption of circulation by ethanol promotes fetal alcohol spectrum disorder (FASD) in medaka (Oryzias latipes) embryogenesis

Khan

Dasmahapatra

2008

Comparative Biochemistry and Physiology Part C: Toxicology &

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Japanese medaka (Oryzias latipes) embryos exposed to ethanol have developed craniofacial, cardiovascular and skeletal defects which can be compared with the phenotypic features of fetal alcohol spectrum disorder (FASD) observed in human. The present experiment was designed to show that the disruption in circulation by ethanol during embryogenesis is a potential cause of FASD. Fertilized eggs were exposed to ethanol (0, 100 and/or 400 mM) for 24 or 48 h at various developmental stages (Iwamatsu stages 4-30) and were analyzed at 6 day post fertilization (dpf). It was observed that controls and the embryos exposed to 100 mM ethanol were in circulating state; however, a significant number of embryos of stages 4-24 exposed to 400 mM ethanol had disrupted circulation. Compared to controls, protein and RNA contents were significantly reduced in noncirculating embryos. Lipid peroxidation (LPO) analysis was made at 3, 6, 24, 48, 96 and 144 hour post fertilization (hpf). LPO was increased with the advancement of morphogenesis; however, ethanol or the circulation status had no effect. We further analyzed alcohol dehydrogenase (Adh 5 and adh8) and aldehyde dehydrogenase (Aldh9A and Aldh1A2) enzyme mRNAs in the embryos exposed to 400 mM ethanol for 24h. A developmental stage specific reduction in these enzyme mRNAs by ethanol was observed. We conclude that ethanol-induced disruption in circulation during embryogenesis is a potential cause of the development of FASD features in medaka.

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Ethanol effects on the developing zebrafish: neurobehavior and skeletal morphogenesis

Cited by 238 publications

References 80 publications

Japanese medaka (Oryzias latipes): developmental model for the study of alcohol teratology

Japanese medaka (Oryzias latipes): developmental model for the study of alcohol teratology

Non-Associative Learning in Larval Zebrafish

Disruption of circulation by ethanol promotes fetal alcohol spectrum disorder (FASD) in medaka (Oryzias latipes) embryogenesis

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