The objective is to create a global gene expression atlas of craniofacial development. The central thesis is that a combination of laser capture microdissection, and FACS, combined with microarrays can be used to efficiently achieve this goal. Microarrays with essentially complete gene representation can be used to rapidly determine the expression levels of every gene in laser capture microdissected elements of craniofacial development. A single experiment, therefore, provides a comprehensive analysis of the gene expression status of one component and a limited number of experiments examining each structure and cell type can create an atlas. A combination of structure, lectin staining, and transgenic GFP expression will be used to precisely identify specific compartments and lineages, including cells driving neural crest induction, nasal placodes and pits, lateral and medial facial eminences, neural crest and paraxial mesoderm cells, maxilla and mandibular recesses, signaling centers, and the structures of palatogenesis.
Specific Aim 2, more limited in scope, is to make Sp8-GFP-Cre and Fgf8-GFP-Cre transgenic mouse tools, which will serve a dual purpose, to allow identification of additional discrete craniofacial components for Specific Aims 1 and 3, and to aid future domain specific gene knockout studies by the craniofacial research community.
Specific Aim 3 is to characterize the Sp8 mutant mouse, which shows a massive disruption of craniofacial development. We hypothesize that Sp8 is upstream of FGF signaling first in the medial neuroepithelium of the forebrain, important for driving neural crest proliferation and migration, and later in the nasal placodes and pits, important for signaling the forming facial eminences. We propose a combined molecular marker analysis;a Cre based genetic dissection of Sp8 function, and a microarray based gene expression profile comparison of signaling centers in wild type and Sp8 mutants.

Public Health Relevance

Cleft lip and palate are very common birth defects, and there are many other facial malformations that are more severe, but less common. These problems have both genetic and environmental causes. We propose to use the latest technologies to study all of the genes involved in making the face, to gain a better understanding of this complicated process. It is hoped this work will lead to new methods to prevent and/or treat these birth defects.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01DE020049-03
Application #
8055994
Study Section
Special Emphasis Panel (ZDE1-JH (24))
Program Officer
Scholnick, Steven
Project Start
2009-09-21
Project End
2014-04-30
Budget Start
2011-05-01
Budget End
2012-04-30
Support Year
3
Fiscal Year
2011
Total Cost
$277,481
Indirect Cost
Name
Cincinnati Children's Hospital Medical Center
Department
Type
DUNS #
071284913
City
Cincinnati
State
OH
Country
United States
Zip Code
45229
Kousa, Youssef A; Mansour, Tamer A; Seada, Haitham et al. (2017) Shared molecular networks in orofacial and neural tube development. Birth Defects Res 109:169-179
Potter, Andrew S; Potter, S Steven (2015) Molecular Anatomy of Palate Development. PLoS One 10:e0132662
Wu, Chao; Bardes, Eric E; Jegga, Anil G et al. (2014) ToppMiR: ranking microRNAs and their mRNA targets based on biological functions and context. Nucleic Acids Res 42:W107-13
Brunskill, Eric W; Potter, Andrew S; Distasio, Andrew et al. (2014) A gene expression atlas of early craniofacial development. Dev Biol 391:133-46
Kasberg, Abigail D; Brunskill, Eric W; Steven Potter, S (2013) SP8 regulates signaling centers during craniofacial development. Dev Biol 381:312-23
Potter, S Steven; Brunskill, Eric W (2012) Laser capture. Methods Mol Biol 886:211-21
Hochheiser, Harry; Aronow, Bruce J; Artinger, Kristin et al. (2011) The FaceBase Consortium: a comprehensive program to facilitate craniofacial research. Dev Biol 355:175-82
Sardana, Divya; Vasa, Suresh; Vepachedu, Nishanth et al. (2010) PhenoHM: human-mouse comparative phenome-genome server. Nucleic Acids Res 38:W165-74
Kaimal, Vivek; Bardes, Eric E; Tabar, Scott C et al. (2010) ToppCluster: a multiple gene list feature analyzer for comparative enrichment clustering and network-based dissection of biological systems. Nucleic Acids Res 38:W96-102