Epidemiological and laboratory evidence indicates that environmental contaminants such as TCDD affect vertebrate heart development. The receptor for TCDD is the Aryl Hydrocarbon Receptor (AHR), a ligand-activated transcriptional activator. We have used zebrafish, an ideal system for studying heart development in vivo to document the cardiotoxic response to TCDD. These results support a model in which TCDD disrupts normal gene regulation in the developing heart. Our microarray experiments have defined the pattern of gene expression changes produced by TCDD in zebrafish embryonic and adult hearts. We find that AHR activation regulates a different set of genes in embryonic heart cells than in juvenile heart cells. Adult hearts are resistant to TCDD cardiotoxicity, except when regenerating lost tissue after surgical wounding. The sensitivity to TCDD in the embryonic and regenerating adult heart correlate with periods in which epicardial cell migration, division, and differentiation are critical to the heart. This suggests that TCDD may be affecting epicardial cell function. We have developed methods for transplantation of cells from embryonic zebrafish hearts into the wounded adult muscle. These embryonic cells become incorporated into the wounded heart muscle. This project is focused on the following questions: 7 Are the cardiotoxic effects of TCDD mediated by AHR activation in the heart? This will be addressed by using transgenic zebrafish to manipulate AHR activity specifically in the heart. 7 Does TCDD alter epicardial function in the embryonic and regenerating adult heart? This will be addressed by determining whether TCDD blocks epicardial cell formation and migration in the embryonic heart. In addition, we will determine whether TCDD alters the activation, migration, growth, and differentiation of embryonic-like epicardial cells in the adult heart in response to wounding. We will compare the genes affected by TCDD in adult wounded hearts to those affected by TCDD in embryonic hearts. Finally, we will determine whether TCDD prevents transplantation of embryonic heart cells into wounded zebrafish myocardium. 7 Why does TCDD activation of AHR affect a different set of genes in the embryonic heart than in the juvenile heart or the developing jaw? We will test the hypothesis that the AHR complex is different in different tissues, producing different DNA binding site specificity in different tissues. As an alternative, we will test the hypothesis that the chromatin structure at a given gene may allow AHR binding in the embryonic heart, but in other tissues the chromatin structure at this gene would be different, preventing AHR binding. This will be tested using ChIP assays coupled to microarrays (ChIP-chip assays).

Public Health Relevance

Heart defects are among the most common birth defects in humans, and recent epidemiological and laboratory research suggest that heart defects may be caused by exposure to chemicals such as 2,3,7,8-tetrachlorodibenzo-p-dioxin, and polychlorinated biphenyls, commonly known as dioxin and PCBs respectively. This project will be based on the zebrafish, Danio rerio, an important non-human model system widely used for studying heart development. The work will identify mechanisms by which toxic compounds disrupt heart development, and explores the possibility that these compounds alter not only epicardial cell function in the developing heart, but also block regeneration and transplantation of cells into a wounded adult heart.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES012716-08
Application #
8450225
Study Section
Cardiovascular Differentiation and Development Study Section (CDD)
Program Officer
Nadadur, Srikanth
Project Start
2003-12-01
Project End
2014-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
8
Fiscal Year
2013
Total Cost
$369,533
Indirect Cost
$112,088
Name
University of Wisconsin Madison
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Baker, Tracie R; Peterson, Richard E; Heideman, Warren (2014) Using zebrafish as a model system for studying the transgenerational effects of dioxin. Toxicol Sci 138:403-11
Baker, Tracie R; King-Heiden, Tisha C; Peterson, Richard E et al. (2014) Dioxin induction of transgenerational inheritance of disease in zebrafish. Mol Cell Endocrinol 398:36-41
Lanham, Kevin A; Plavicki, Jessica; Peterson, Richard E et al. (2014) Cardiac myocyte-specific AHR activation phenocopies TCDD-induced toxicity in zebrafish. Toxicol Sci 141:141-54
Hofsteen, Peter; Mehta, Vatsal; Kim, Min-Sik et al. (2013) TCDD inhibits heart regeneration in adult zebrafish. Toxicol Sci 132:211-21
Baker, Tracie R; Peterson, Richard E; Heideman, Warren (2013) Early dioxin exposure causes toxic effects in adult zebrafish. Toxicol Sci 135:241-50
Plavicki, Jessica; Hofsteen, Peter; Peterson, Richard E et al. (2013) Dioxin inhibits zebrafish epicardium and proepicardium development. Toxicol Sci 131:558-67
Hofsteen, Peter; Plavicki, Jessica; Johnson, Shaina D et al. (2013) Sox9b is required for epicardium formation and plays a role in TCDD-induced heart malformation in zebrafish. Mol Pharmacol 84:353-60
Lanham, Kevin A; Peterson, Richard E; Heideman, Warren (2012) Sensitivity to dioxin decreases as zebrafish mature. Toxicol Sci 127:360-70
King-Heiden, Tisha C; Mehta, Vatsal; Xiong, Kong M et al. (2012) Reproductive and developmental toxicity of dioxin in fish. Mol Cell Endocrinol 354:121-38
Lanham, Kevin A; Prasch, Amy L; Weina, Kasia M et al. (2011) A dominant negative zebrafish Ahr2 partially protects developing zebrafish from dioxin toxicity. PLoS One 6:e28020

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