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![Ethanol interacts with vangl2 during early embryogenesis. a Quantification of the distance between the lenses in embryos treated with 1% ethanol for 24 h, initiating at four different stages [3.3, 4, 4.5, and 6 h post fertilization (hpf)]. Statistical significance of differences between groups is indicated with compact letter display. Groups with different letters are significantly different from one another (p < 0.05; Tukey’s honest test). Sample size and p values provided in Additional file 9: Table S2. b Percentage of embryos with complete cyclopia following each treatment. Heterozygous embryos were most sensitive to ethanol when exposed from 3.3 to 27.3 hpf. Mutants were most sensitive from 6 to 30 hpf](publication/352900549/figure/fig1/AS:1040991377190913@1625203093716/Ethanol-interacts-with-vangl2-during-early-embryogenesis-a-Quantification-of-the.png)
Ethanol interacts with vangl2 during early embryogenesis. a Quantification of the distance between the lenses in embryos treated with 1% ethanol for 24 h, initiating at four different stages [3.3, 4, 4.5, and 6 h post fertilization (hpf)]. Statistical significance of differences between groups is indicated with compact letter display. Groups with different letters are significantly different from one another (p < 0.05; Tukey’s honest test). Sample size and p values provided in Additional file 9: Table S2. b Percentage of embryos with complete cyclopia following each treatment. Heterozygous embryos were most sensitive to ethanol when exposed from 3.3 to 27.3 hpf. Mutants were most sensitive from 6 to 30 hpf
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Background
Gene-environment interactions are likely to underlie most human birth defects. The most common known environmental contributor to birth defects is prenatal alcohol exposure. Fetal alcohol spectrum disorders (FASD) describe the full range of defects that result from prenatal alcohol exposure. Gene-ethanol interactions underlie susceptibil...
Citations
... The Hh signaling pathway is causally implicated downstream from EtOH in FASD in vertebrate embryos (Abramyan, 2019;Eberhart and Parnell, 2016;Hong and Krauss, 2017;Li et al., 2007;Sidik et al., 2021). Since Hh signaling is also implicated in sea urchin skeletal patterning (Walton et al., 2009), we next asked whether perturbed Hh signaling can explain the EtOH-mediated defects by testing whether the Hh pathway inhibitor cyclopamine or agonist SAG (Chen et al., 2002a(Chen et al., , 2002bFrank-Kamenetsky et al., 2002) can rescue EtOH-treated embryos. ...
... Previous studies have linked FASD abnormalities, both neural and facial, to perturbations of the Hh and RA signaling pathways (Ahlgren et al., 2002;Cayuso et al., 2006;Cohen and Sulik, 1992;Petrelli et al., 2019;Reimers et al., 2004;Smith et al., 2014), each of which plays important roles in the development of both the CNS (Chatzi et al., 2013;Janesick et al., 2015;Li et al., 2021) and the facial skeleton (Abramyan, 2019;Dubey et al., 2018;Gur et al., 2022;Sun et al., 2020). Experiments in zebrafish (Marrs et al., 2010) and frogs (Johnson et al., 2007a;Yelin et al., 2005) demonstrated that exogenous RA suffices to rescue EtOH-mediated neural and skeletal defects; similarly, addition of the Hh pathway agonist rescues development of EtOH-exposed zebrafish (Sidik et al., 2021). While the precise mechanism of these effects remains unclear, embryonic exposure to EtOH can induce holoprosencephaly, strongly implicating an inhibitory effect on Shh signaling (Hong and Krauss, 2017); in keeping with this, an ethanol-mediated blockade to Shh protein processing has been reported (Li et al., 2007). ...
Ethanol is a known vertebrate teratogen that causes craniofacial defects as a component of fetal alcohol syndrome (FAS). Our results show that sea urchin embryos treated with ethanol similarly show broad skeletal patterning defects, potentially analogous to the defects associated with FAS. The sea urchin larval skeleton is a simple patterning system that involves only two cell types: the primary mesenchymal cells (PMCs) that secrete the calcium carbonate skeleton and the ectodermal cells that provide migratory, positional, and differentiation cues for the PMCs. Perturbations in RA biosynthesis and Hh signaling pathways are thought to be causal for the FAS phenotype in vertebrates. Surprisingly, our results indicate that these pathways are not functionally relevant for the teratogenic effects of ethanol in developing sea urchins. We found that developmental morphology as well as the expression of some ectodermal and PMC genes was delayed by ethanol exposure. Temporal transcriptome analysis revealed significant impacts of ethanol on signaling and metabolic gene expression, and a disruption in the timing of GRN gene expression that includes both delayed and precocious gene expression throughout the specification network. We conclude that the skeletal patterning perturbations in ethanol-treated embryos likely arise from a loss of temporal synchrony within and between the instructive and responsive tissues.
... FASD include Fetal Alcohol Syndrome (FAS), partial Fetal Alcohol Syndrome (pFAS), alcohol-related neurodevelopmental disorders (ARND), and alcohol-related birth defects (ARBD) [1]. FAS is believed to be the most common teratogenically induced, nonhereditary form of mental deficiency throughout the Western world [2][3][4]. It is associated with reduced intelligence, attention disorders, neuropsychological deficits, physical abnormalities including facial dysmorphology, sleep disorders, and behavioral problems [5][6][7]. ...
Fetal alcohol spectrum disorders (FASD) in a course of high prenatal alcohol exposure (hPAE) are among the most common causes of developmental disorders. The main reason for pharmacological treatment of FASD children is attention deficit hyperactivity disorder (ADHD), and methylphenidate (MPH) is the drug of choice. The aim of the study was to assess whether children born of hPAE with ADHD, with or without morphological FASD, differ in terms of catechol-O-methyltransferase (COMT) and dopamine receptor D2 (DRD2) gene polymorphisms, and if genetic predisposition affects response and safety of MPH treatment. The polymorphisms of COMT (rs4680) and DRD2 (rs1076560, rs1800497) were analyzed in DNA samples. A borderline significance was found for the correlation between MPH side effects and the G allele of COMT (rs4680) (p = 0.04994) in all ADHD children. No effect of COMT (rs4680) and DRD2 (rs1076560, rs1800497) polymorphisms and the treatment efficacy was observed. The analyzed DRD2 and COMT gene polymorphisms seem to play no role in MPH efficacy in ADHD children with hPAE, while low-activity COMT (Met158) variant carriers may be more intolerant to MPH. The MPH treatment is effective in ADHD independent of FASD, although the ADHD-FASD variant requires higher doses to be successful. These results may help in optimization and individualization in child psychiatry.
... For example, RA regulates components of the Wnt pathway (Zhao and Duester, 2009;Carron and Shi, 2016) and RA target genes interact with components of this pathway (Harada et al., 2007;Zhang et al., 2013). Ethanol similarly interacts with the Wnt/PCP pathway (Sarmah et al., 2020;Sidik et al., 2021). These observations suggest that the RA effects on gastrulation and the rostral movement of the LEM/PCM involve an interaction with the Wnt/PCP pathway to regulate morphogenetic movements. ...
Retinoic acid (RA) is a central regulatory signal that controls numerous developmental processes in vertebrate embryos. Although activation of Hox expression is considered one of the earliest functions of RA signaling in the embryo, there is evidence that embryos are poised to initiate RA signaling just before gastrulation begins, and manipulations of the RA pathway have been reported to show gastrulation defects. However, which aspects of gastrulation are affected have not been explored in detail. We previously showed that partial inhibition of RA biosynthesis causes a delay in the rostral migration of some of the earliest involuting cells, the leading edge mesendoderm (LEM) and the prechordal mesoderm (PCM). Here we identify several detrimental gastrulation defects resulting from inhibiting RA biosynthesis by three different treatments. RA reduction causes a delay in the progression through gastrulation as well as the rostral migration of the goosecoid-positive PCM cells. RA inhibition also hampered the elongation of explanted dorsal marginal zones, the compaction of the blastocoel, and the length of Brachet’s cleft, all of which indicate an effect on LEM/PCM migration. The cellular mechanisms underlying this deficit were shown to include a reduced deposition of fibronectin along Brachet’s cleft, the substrate for their migration, as well as impaired separation of the blastocoel roof and involuting mesoderm, which is important for the formation of Brachet’s cleft and successful LEM/PCM migration. We further show reduced non-canonical Wnt signaling activity and altered expression of genes in the Ephrin and PDGF signaling pathways, both of which are required for the rostral migration of the LEM/PCM, following RA reduction. Together, these experiments demonstrate that RA signaling performs a very early function critical for the progression of gastrulation morphogenetic movements.
... Mutation of vangl2 sensitizes the zebrafish to craniofacial anomalies induced by blebbistatin, an inhibitor of the Wnt/ PCP pathway (Sidik et al., 2021). Genes involved in the PCP pathway have also emerged as susceptibility-inducing genes in ASD and other DDs (Sans et al., 2016;Milgrom-Hoffman and Humbert, 2018). ...
Various genetic and environmental factors are associated with developmental disorders (DDs). It has been suggested that interaction between genetic and environmental factors (G × E) is involved in the etiology of DDs. There are two major approaches to analyze the interaction: genome-wide and candidate gene-based approaches. In this mini-review, we demonstrate how these approaches can be applied to reveal the G × E related to DDs focusing on zebrafish and mouse models. We also discuss novel approaches to analyze the G × E associated with DDs.
... Work in Xenopus and zebrafish show that the Wnt/PCP pathway plays a critical role in regulating early gastrulation cell movements, suggesting that ethanol is disrupting cell signaling during these cell movements (Hardy et al., 2008;Heisenberg et al., 2000;Ohkawara, 2003;Roszko et al., 2009;Ulrich, 2003). Recent work from zebrafish has shown that Wnt/PCP members vangl2 and gcp4 interact with ethanol during gastrulation disrupting convergence extension leading to cyclopia, an extreme form of holoprosencephaly (Sidik et al., 2021;Swartz et al., 2014). ...
... More recent work from the Eberhart lab shows that ethanol exposure in vangl2 mutants disrupts the number and polarity of cell protrusions. Strikingly, this ethanol exposure does not impact expression of PCP members, but acts indirectly through localization of shha expression (Sidik et al., 2021). This work demonstrates ethanol can interact with and disrupt multiple pathways underlying FASD, though human vangl2 has yet to be implicated in FASD. ...
Fetal alcohol spectrum disorder (FASD) describes a wide range of structural deficits and cognitive impairments. FASD impacts up to 5% of children born in the United States each year, making ethanol one of the most common teratogens. Due to limitations and ethical concerns, studies in humans are limited in their ability to study FASD. Animal models have proven critical in identifying and characterizing the mechanisms underlying FASD. In this review, we will focus on the attributes of zebrafish that make it a strong model in which to study ethanol-induced developmental defects. Zebrafish have several attributes that make it an ideal model in which to study FASD. Zebrafish produced large numbers of externally fertilized, translucent embryos. With a high degree of genetic amenability, zebrafish are at the forefront of identifying and characterizing the gene-ethanol interactions that underlie FASD. Work from multiple labs has shown that embryonic ethanol exposures result in defects in craniofacial, cardiac, ocular, and neural development. In addition to structural defects, ethanol-induced cognitive and behavioral impairments have been studied in zebrafish. Building upon these studies, work has identified ethanol-sensitive loci that underlie the developmental defects. However, analyses show there is still much to be learned of these gene-ethanol interactions. The zebrafish is ideally suited to expand our understanding of gene-ethanol interactions and their impact on FASD. Because of the conservation of gene function between zebrafish and humans, these studies will directly translate to studies of candidate genes in human populations and allow for better diagnosis and treatment of FASD.
As an important part in international disease, mental disorders seriously damage human health and social stability, which show the complex pathogenesis and increasing incidence year by year. In order to analyze the pathogenesis of mental disorders as soon as possible and to look for the targeted drug treatment for psychiatric diseases, a more reasonable animal model is imperious demands. Benefiting from its high homology with the human genome, its brain tissue is highly similar to that of humans, and it is easy to realize whole-body optical visualization and high-throughput screening; zebrafish stands out among many animal models of mental disorders. Here, valuable qualified zebrafish mental disorders models could be established through behavioral test and sociological analysis, which are simulated to humans, and combined with molecular analyses and other detection methods. This review focuses on the advances in the zebrafish model to simulate the human mental disorders; summarizes the various behavioral characterization means, the use of equipment, and operation principle; sums up the various mental disorder zebrafish model modeling methods; puts forward the current challenges and future development trend, which is to contribute the theoretical supports for the exploration of the mechanisms and treatment strategies of mental disorders.
The zebrafish ( Danio rerio ), is an important biomedical model organism used in many disciplines, including development, disease modeling and toxicology, to better understand vertebrate biology. The phenomenon of developmental delay in zebrafish embryos has been widely reported as part of a mutant or treatment-induced phenotype, and accurate characterization of such delays is imperative. Despite this, the only way at present to identify and quantify these delays is through manual observation, which is both time-consuming and subjective. Machine learning approaches in biology are rapidly becoming part of the toolkit used by researchers to address complex questions. In this work, we introduce a machine learning-based classifier that has been trained to detect temporal developmental differences across groups of zebrafish embryos. Our classifier is capable of rapidly analyzing thousands of images, allowing comparisons of developmental temporal rates to be assessed across and between experimental groups of embryos. Finally, as our classifier uses images obtained from a standard live-imaging widefield microscope and camera set-up, we envisage it will be readily accessible to the zebrafish community, and prove to be a valuable resource.
The zebrafish ( Danio rerio ), is an important biomedical model organism used in many disciplines, including development, disease modeling and toxicology, to better understand vertebrate biology. The phenomenon of developmental delay in zebrafish embryos has been widely reported as part of a mutant or treatment-induced phenotype, and accurate characterization of such delays is imperative. Despite this, the only way at present to identify and quantify these delays is through manual observation, which is both time-consuming and subjective. Machine learning approaches in biology are rapidly becoming part of the toolkit used by researchers to address complex questions. In this work, we introduce a machine learning-based classifier that has been trained to detect temporal developmental differences across groups of zebrafish embryos. Our classifier is capable of rapidly analyzing thousands of images, allowing comparisons of developmental temporal rates to be assessed across and between experimental groups of embryos. Finally, as our classifier uses images obtained from a standard live-imaging widefield microscope and camera set-up, we envisage it will be readily accessible to the zebrafish community, and prove to be a valuable resource.
Birth defects are relatively common congenital outcomes that significantly impact affected individuals, their families, and communities. Effective development and deployment of prevention and therapeutic strategies for these conditions requires sufficient understanding of etiology, including underlying genetic and environmental causes. Tremendous progress has been made in defining the genetic basis of familial and syndromic forms of birth defects. However, the majority of birth defect cases are considered nonsyndromic and thought to result from multifactorial gene-environment interactions. While substantial advances have been made in elucidating the genetic landscape of these etiologically complex conditions, significant biological and technical constraints have stymied progress toward a refined knowledge of environmental risk factors. Defining specific gene-environment interactions in birth defect etiology is even more challenging. However, progress has been made, including demonstration of critical proofs of concept and development of new conceptual and technical approaches for resolving complex gene-environment interactions. In this review, we discuss current views of multifactorial birth defect etiology, comparing them with other diseases that also involve gene-environment interactions, including primary immunodeficiency and cancer. We describe how various model systems have illuminated mechanisms of multifactorial etiology and these models’ individual strengths and weaknesses. Finally, suggestions for areas of future emphasis are proposed.
Most human birth defects are thought to result from complex interactions between combinations of genetic and environmental factors. This is true even for conditions that, at face value, may appear simple and straightforward, like fetal alcohol spectrum disorders (FASD). FASD describe the full range of structural and neurological disruptions that result from prenatal alcohol exposure. While FASD require alcohol exposure, evidence from human and animal model studies demonstrate that additional genetic and/or environmental factors can influence the embryo's susceptibility to alcohol. Only a limited number of alcohol interactions in birth defects have been identified, with many sensitizing genetic and environmental factors likely yet to be identified. Because of this, while unsatisfying, there is no definitively “safe” dose of alcohol for all pregnancies. Determining these other factors, as well as mechanistically characterizing known interactions, is critical for better understanding and preventing FASD and requires combined scrutiny of human and model organism studies.
