Craniofacial abnormalities are characteristic of embryonic exposure to alcohol. In typical fetal alcohol syndrome poor development of a number of facial features, all of which are derived from the cranial neural crest, is observed. This grant seeks to establish a zebrafish model to study the craniofacial defects associated with embryonic alcohol exposure, and to establish that such environmental insults produce craniofacial defects by interfering with normal signals that control growth. We will first establish what the optimal dose and time schedule for alcohol application is, with respect to deficits in cranial neural crest cells, taking into consideration doses which would be relevant to fetal alcohol syndrome. The zebrafish is an excellent model for comparing teratogens, like alcohol, with genetic defects, to determine what candidate genes might be altered by environmental conditions. In addition, it is possible to overexpress genes of interest by directed injection or transgenesis. The defects in fetal alcohol syndrome partially overlap with some features of holoprosencephaly, which is typically a more dramatic malformation of the central nervous system with associated midline facial features, arising from both genetic and environmental factors. One such genetic cause is a heterozygous mutation in the human Sonic Hedgehog gene. Partial inhibition of Sonic hedgehog in the chick embryo using function-blocking antibodies results in a phenotype similar to a mild holoprosencephaly that is intriguingly similar to fetal alcohol syndrome. The morphological similarity between embryos exposed to alcohol and partial inhibition of Sonic hedgehog suggests a potential mechanistic link. Zebrafish have multiple hedgehog genes, so it is possible that alcohol interferes with a common signaling pathway. To test the hypothesis that ethanol leads to cranial neural crest cell death via a decrease in the availability of zebrafish hedgehog genes, the experiments described in this grant will examine the effect of ethanol on the message levels of genes in the hedgehog signaling pathway. We will also examine the fate of cranial neural crest cells, as well as the growth of the craniofacial structures. We will further attempt to rescue the cranial neural crest cells after ethanol treatment by application of exogenous Sonic hedgehog. These experiments will lay the groundwork to expand our understanding of the mechanisms of craniofacial defects following embryonic exposure to alcohol.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AA013596-01
Application #
6460354
Study Section
Special Emphasis Panel (ZAA1-CC (01))
Program Officer
Neuhold, Lisa
Project Start
2002-09-30
Project End
2003-08-31
Budget Start
2002-09-30
Budget End
2003-08-31
Support Year
1
Fiscal Year
2002
Total Cost
$160,000
Indirect Cost
Name
California Institute of Technology
Department
Type
Schools of Arts and Sciences
DUNS #
078731668
City
Pasadena
State
CA
Country
United States
Zip Code
91125
Loucks, Evyn; Ahlgren, Sara (2012) Assessing teratogenic changes in a zebrafish model of fetal alcohol exposure. J Vis Exp :
Schwend, Tyler; Loucks, Evyn J; Snyder, Diana et al. (2011) Requirement of Npc1 and availability of cholesterol for early embryonic cell movements in zebrafish. J Lipid Res 52:1328-44
Schwend, Tyler; Loucks, Evyn J; Ahlgren, Sara C (2010) Visualization of Gli activity in craniofacial tissues of hedgehog-pathway reporter transgenic zebrafish. PLoS One 5:e14396
Loucks, Evyn J; Ahlgren, Sara C (2009) Deciphering the role of Shh signaling in axial defects produced by ethanol exposure. Birth Defects Res A Clin Mol Teratol 85:556-67
Schwend, Tyler; Ahlgren, Sara C (2009) Zebrafish con/disp1 reveals multiple spatiotemporal requirements for Hedgehog-signaling in craniofacial development. BMC Dev Biol 9:59
Loucks, Evyn J; Schwend, Tyler; Ahlgren, Sara C (2007) Molecular changes associated with teratogen-induced cyclopia. Birth Defects Res A Clin Mol Teratol 79:642-51