Deciphering the role of Shh signaling in axial defects produced by ethanol exposure.
BACKGROUND
The phenotype of embryos exposed to ethanol is complex and likely due to multiple alterations in developmental pathways. We have previously demonstrated that Sonic hedgehog signaling (Shh-s) was reduced in both chicken and zebrafish embryos when exposed to ethanol.


METHODS
There are many tissues affected by embryonic ethanol exposure, and in this article we explore the development of axial tissues, using zebrafish embryos. We then compare these effects to the phenotypes produced by exposure to two drugs that also inhibit Shh-s: cyclopamine and forskolin.


RESULTS
We found alterations in the development of the notochord and somites produced by all three compounds, although only ethanol produced developmental delay of epiboly. Upon observation of early developing embryos, muscle pioneer cells were completely lost in cyclopamine-treated embryos, and reduced, but less so, in embryos treated with forskolin and ethanol. Ethanol treatment produced a dose-dependent reduction in total body length that may be linked to epiboly delay seen earlier during development. Despite the differences between cyclopamine and forskolin, we found that shh mRNA injection rescued the short body length, the alteration in somite shape, and the cyclopia produced by ethanol exposure.


CONCLUSIONS
Taken together, each teratogen produced a unique set of phenotypic changes in the body axis, suggesting that each compound affects Shh-s and also produces a distinctive set of molecular alterations. However, addition of exogenous Shh to ethanol treated zebrafish prevented many of the gross physical phenotypes, suggesting that the suppression of Shh-s is one of the major effects of ethanol exposure.
Loucks
Evyn J
EJ
Children's Memorial Research Center Program in Developmental Biology, Chicago, Illinois 60640, USA.
Ahlgren
Sara C
SC
D000818:Animals; D002492:Central Nervous System Depressants / Q000494:pharmacology / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects*; D000431:Ethanol / Q000494:pharmacology / Q000633:toxicity*; D053823:Hedgehog Proteins / Q000378:metabolism*; D009672:Notochord / Q000196:embryology*; D015398:Signal Transduction / Q000187:drug effects*; D013723:Teratogens / Q000494:pharmacology / Q000633:toxicity*; D015027:Zebrafish / Q000196:embryology*; D029961:Zebrafish Proteins / Q000378:metabolism*
D002492:Central Nervous System Depressants; D053823:Hedgehog Proteins; D013723:Teratogens; D029961:Zebrafish Proteins; D000431:Ethanol