title: Molecular analysis of coronal perisutural tissues in a craniosynostotic rabbit model using polymerase chain reaction suppression subtractive hybridization.
abstract: BACKGROUND
  In the United States, the incidence of craniosynostosis (premature fusion of the sutures of the cranial vault) is one in 2000 to 3000 live births. The condition can cause increased intracranial pressure, severely altered head shape, and mental retardation. The authors have previously described a colony of rabbits with heritable coronal suture synostosis. This model has been instrumental in describing the postsurgical craniofacial growth associated with craniosynostosis. The molecular analysis of this model has been limited by the lack of molecular tools for use in rabbits. To understand the pathogenesis of craniosynostosis, the authors compared gene expression in perisutural tissues between wild-type and craniosynostotic rabbits using polymerase chain reaction suppression subtractive hybridization.
  
  
  METHODS
  Suppression subtractive hybridization polymerase chain reaction was performed on RNA derived from pooled samples of calvariae from 10-day-old wild-type (n = 3) and craniosynostotic (n = 3) rabbits to obtain cDNA clones enriched in either wild-type tissues (underexpressed in craniosynostotic tissue) or craniosynostotic tissues (overexpressed in craniosynostotic compared with wild-type).
  
  
  RESULTS
  Differential expression was identified for approximately 140 recovered cDNA clones up-regulated in craniosynostotic tissues and 130 recovered clones for wild-type tissues. Of these, four genes were confirmed by quantitative reverse-transcriptase polymerase chain reaction as being overexpressed in craniosynostotic sutural tissue: β-globin (HBB), osteopontin (SPP1), osteonectin (SPARC), and cathepsin K (CTSK). Two genes were confirmed to be underexpressed in the craniosynostotic samples: collagen 3, alpha 1 (COL3A1) and ring finger protein 12 (RNF12).
  
  
  CONCLUSIONS
  The differential expression of these gene products in our naturally occurring craniosynostotic model appears to be the result of differences in the normal bone formation/resorption pathway.
authors.affiliation: Pittsburgh and Philadelphia, Pa. From the Department of Surgery, Division of Plastic and Reconstructive Surgery, University of Pittsburgh and Pediatric Craniofacial Biology Laboratory, Children's Hospital of Pittsburgh; the Center for Genomic Sciences, Allegheny-Singer Research Institute, West Penn Allegheny Health Systems; the Departments of Anthropology, Orthodontics, Oral Biology, and Bioengineering, University of Pittsburgh; the Division of Plastic Surgery, Allegheny General Hospital of Pittsburgh; and the Department of Microbiology and Immunology, Drexel University College of Medicine.
authors.forename: James J
authors.initials: JJ
authors.lastname: Cray
authors.forename: Phillip H
authors.initials: PH
authors.lastname: Gallo
authors.forename: Emily L
authors.initials: EL
authors.lastname: Durham
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Losee
authors.forename: Mark P
authors.initials: MP
authors.lastname: Mooney
authors.forename: Sandeep
authors.initials: S
authors.lastname: Kathju
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Cooper
mesh_terms: D000818:Animals; D003398:Craniosynostoses / Q000235:genetics*; D004195:Disease Models, Animal; D018507:Gene Expression Regulation, Developmental; D016133:Polymerase Chain Reaction / Q000379:methods*; D011817:Rabbits