Zebrafish have become a powerful new model organism for studies of vertebrate development. This is largely due to the optical clarity of zebrafish embryos and the advent of green fluorescent protein (GFP) transgenic technology. Combined, these features allow the characterization of organogenesis at the level of individual cells. Additionally, developmental genetic regulatory pathways have been found to be highly conserved between zebrafish and mammals. We first propose to exploit the advantages of this system to characterize the detailed normal morphogenesis of zebrafish teeth by the timelapse confocal microscopy of recently-generated GFP transgenic lines. We will use a Fli1:GFP line to specifically follow cells in the dental mesenchyme, a Shh:GFP line to trace the movements of the dental epithelium, and the histone H2A:GFP line to follow the nuclei of each cell in the tooth germ. We also propose to generate new transgenic zebrafish lines expressing GFP and DsRed driven by Fgf4 regulatory elements. We will use these lines to investigate the presence of a zebrafish homolog of the mammalian enamel knot, an organizing center within the mammalian tooth epithelium that expresses Fgf4. Lastly we propose to perturb the normal development of teeth with the FGF receptor inhibitor Su5402 and observe the effects on morphogenesis with the GFP lines mentioned. These studies will reveal cell behavior during normal and experimentally perturbed tooth morphogenesis at a level of detail never before described. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
1F32DE015029-01A1
Application #
6692774
Study Section
NIDCR Special Grants Review Committee (DSR)
Program Officer
Hardwick, Kevin S
Project Start
2003-06-01
Project End
2006-05-31
Budget Start
2003-06-01
Budget End
2004-05-31
Support Year
1
Fiscal Year
2003
Total Cost
$46,420
Indirect Cost
Name
University of Colorado at Boulder
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
007431505
City
Boulder
State
CO
Country
United States
Zip Code
80309
Jackman, William R; Davies, Shelby H; Lyons, David B et al. (2013) Manipulation of Fgf and Bmp signaling in teleost fishes suggests potential pathways for the evolutionary origin of multicuspid teeth. Evol Dev 15:107-18
Gibert, Yann; Bernard, Laure; Debiais-Thibaud, Melanie et al. (2010) Formation of oral and pharyngeal dentition in teleosts depends on differential recruitment of retinoic acid signaling. FASEB J 24:3298-309
Jackman, William R; Yoo, James J; Stock, David W (2010) Hedgehog signaling is required at multiple stages of zebrafish tooth development. BMC Dev Biol 10:119
Stock, David W; Jackman, William R; Trapani, Josh (2006) Developmental genetic mechanisms of evolutionary tooth loss in cypriniform fishes. Development 133:3127-37
Jackman, William R; Stock, David W (2006) Transgenic analysis of Dlx regulation in fish tooth development reveals evolutionary retention of enhancer function despite organ loss. Proc Natl Acad Sci U S A 103:19390-5
Jackman, William R; Draper, Bruce W; Stock, David W (2004) Fgf signaling is required for zebrafish tooth development. Dev Biol 274:139-57