PUBLICATION
Exposure to ethanol leads to midfacial hypoplasia in a zebrafish model of FASD via indirect interactions with the Shh pathway
- Authors
- Sidik, A., Dixon, G., Buckley, D.M., Kirby, H.G., Sun, S., Eberhart, J.K.
- ID
- ZDB-PUB-210703-37
- Date
- 2021
- Source
- BMC Biology 19: 134 (Journal)
- Registered Authors
- Eberhart, Johann
- Keywords
- Convergent extension, Cyclopia, Ethanol, Fetal alcohol spectrum disorders (FASD), Gastrulation, Polarity, Sonic Hedgehog (Shh), Vangl2, Wnt/planar cell polarity (PCP) pathway
- MeSH Terms
-
- Animals
- Cell Polarity
- Ethanol/toxicity
- Female
- Fetal Alcohol Spectrum Disorders*/genetics
- Hedgehog Proteins/genetics
- Humans
- Pregnancy
- Prenatal Exposure Delayed Effects
- Wnt Signaling Pathway
- Zebrafish*/genetics
- Zebrafish Proteins/genetics
- PubMed
- 34210294 Full text @ BMC Biol.
Citation
Sidik, A., Dixon, G., Buckley, D.M., Kirby, H.G., Sun, S., Eberhart, J.K. (2021) Exposure to ethanol leads to midfacial hypoplasia in a zebrafish model of FASD via indirect interactions with the Shh pathway. BMC Biology. 19:134.
Abstract
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 susceptibility to FASD, but we lack a mechanistic understanding of these interactions. Here, we leverage the genetic tractability of zebrafish to address this problem.
Results We first show that vangl2, a member of the Wnt/planar cell polarity (Wnt/PCP) pathway that mediates convergent extension movements, strongly interacts with ethanol during late blastula and early gastrula stages. Embryos mutant or heterozygous for vangl2 are sensitized to ethanol-induced midfacial hypoplasia. We performed single-embryo RNA-seq during early embryonic stages to assess individual variation in the transcriptional response to ethanol and determine the mechanism of the vangl2-ethanol interaction. To identify the pathway(s) that are disrupted by ethanol, we used these global changes in gene expression to identify small molecules that mimic the effects of ethanol via the Library of Integrated Network-based Cellular Signatures (LINCS L1000) dataset. Surprisingly, this dataset predicted that the Sonic Hedgehog (Shh) pathway inhibitor, cyclopamine, would mimic the effects of ethanol, despite ethanol not altering the expression levels of direct targets of Shh signaling. Indeed, we found that ethanol and cyclopamine strongly, but indirectly, interact to disrupt midfacial development. Ethanol also interacts with another Wnt/PCP pathway member, gpc4, and a chemical inhibitor of the Wnt/PCP pathway, blebbistatin, phenocopies the effect of ethanol. By characterizing membrane protrusions, we demonstrate that ethanol synergistically interacts with the loss of vangl2 to disrupt cell polarity required for convergent extension movements.
Conclusions Our results show that the midfacial defects in ethanol-exposed vangl2 mutants are likely due to an indirect interaction between ethanol and the Shh pathway. Vangl2 functions as part of a signaling pathway that regulates coordinated cell movements during midfacial development. Ethanol exposure alters the position of a critical source of Shh signaling that separates the developing eye field into bilateral eyes, allowing the expansion of the midface. Collectively, our results shed light on the mechanism by which the most common teratogen can disrupt development.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping