Screening Differentially Expressed Genes In Mouse Head Development By Whole Genome Microarray And Verification

ObjectiveTo detect the gene expression profile of mouse head at the stage of neural crest migration(embryonic day 9.5(E9.5))and derivation(E13.5),screen differentially expressed genes,and discuss on the molecular mechanism of mouse cranial neural crest migration and derivation and mouse craniofacial development.MethodsWe use whole mouse genome microarray to detect the gene expression profile in head tissue of mouse E9.5 and E13.5 and then screen differentially expressed genes,carry out GO analysis and Pathway analysis.We use real-time quantitative PCR to detected five significant differences in gene expression,and validate the result of the gene microarray,and we also detect the five genes expression in the mandibular of 1d and 8 week mouse.Results5469 genes were found differentially expressed in the head tissue of mouse E9.5 and E13.5,of which there are 3118 genes up-regulation more than 2 times and 2351 genes down-regulated more than 2 times.These genes were involved in 2960 GO terms and 82 pathways.The pathways in relation to bone development involved in Calcium signaling pathway,MAPK signaling pathway,Wnt signaling pathway,Regulation of actin cytoskeleton,Cell cycle,p53 signaling pathway,etc.The pathways in relation to cell migration involved in Focal adhesion,Cell adhesion molecules(CAMs),ECM-receptor interaction,Gap junction,Tight junction,PI3K-Akt signaling pathway,Rap1 signaling pathway,Erb B signaling pathway,Wnt signaling pathway,p53 signaling pathway,Ras signaling pathway,Cell cycle etc.Through q PCR we found the relative expression of Bmp4,Cdh6,Shh,Sox9,Runx2 were 6.59、0.127、58.03、4.69、97.65.The expression trend of these genes coincides with the genome microarray.ConclusionA number of differently expressed genes were detected by whole mouse genome microarray.The results of GO analysis and Pathway analysis suggest that early embryonic development of mouse head is a complex process regulated by multiple genes and molecule signals.Gene expression of Bmp4,Cdh6,Shh,Sox9,Runx2 verified the reliability of microarray result and suggest Bmp4,Shh,Sox9,Runx2 play an important role in lower jaw development and Cdh6 is necessary in neural crest migration.