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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 are being 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.
Other specific aims are:
Public Health Relevance
Cleft lip and palate are 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 treat these birth defects.
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Normal mouse embryo (left) and Sp8 mutant (right) with the face missing, but tongue and mandible present and normal
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