Quantitative transcriptional analysis of fusing and nonfusing cranial suture complexes in mice.
Previous studies have documented the differences in expression of various genes associated with the process of osteogenesis in fusing and nonfusing cranial sutures, including growth factors, growth factor receptors, and extracellular matrix molecules. Most of these studies were performed in rats, and although the biology regulating cranial suture fusion in mice and rats is presumed to be similar, studies are needed to verify these expression patterns as mice become increasingly utilized for scientific inquiry into the molecular biology of suture fusion and patency. The purpose of this study was to determine the differences in expression of several genes known to be critical to osteoblast biology. Posterior frontal and sagittal suture complexes (including the associated dura mater, suture mesenchyme, and osteogenic fronts) were isolated from 5-, 15-, 25-, 35-, and 45-day-old male CD-1 mice (n = 8 per age; n = 40 total). Total cellular RNA was extracted and converted to cDNA. Quantitative real-time reverse transcriptase polymerase chain reaction was then performed for the following genes: transforming growth factor beta1 and beta3, fibroblast growth factor receptor 1, Runx2,Osteopontin, and Osteocalcin. Expression of all genes examined was increased significantly in the posterior frontal suture as compared with the sagittal suture. Peak expression for all genes was observed on day 25. These data demonstrate that the expression of osteogenic growth factors, growth factor receptors, transcription factors, and extracellular matrix molecules is increased in the fusing posterior frontal suture in mice.
Nacamuli
Randall P
RP
Department of Surgery, Stanford University School of Medicine, Stanford, Calif 94305-5148, USA.
Song
Hanjoon M
HM
Fang
Tony D
TD
Fong
Kenton D
KD
Mathy
Jonathan A
JA
Shi
Yun-Ying
YY
Salim
Ali
A
Longaker
Michael T
MT
D000818:Animals; D050659:Core Binding Factor Alpha 1 Subunit; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000235:genetics*; D015870:Gene Expression; D006133:Growth Substances / Q000235:genetics*; D051379:Mice; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D010012:Osteogenesis / Q000235:genetics*; D053495:Osteopontin; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000235:genetics / Q000378:metabolism; D012886:Skull / Q000378:metabolism*; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D053773:Transforming Growth Factor beta1; D053782:Transforming Growth Factor beta3
D050659:Core Binding Factor Alpha 1 Subunit; D004268:DNA-Binding Proteins; D016326:Extracellular Matrix Proteins; D006133:Growth Substances; D012333:RNA, Messenger; C493716:Runx2 protein, mouse; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; C508961:Tgfb1 protein, mouse; C509045:Tgfb3 protein, mouse; D050656:Transcription Factor AP-2; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D053782:Transforming Growth Factor beta3; D015675:Osteocalcin; D053495:Osteopontin