Analysis Of Gene Expression And Function In The Tongue Of Retinoic Acid-Induced Cleft Palate By Gene-Chip Technology

Background: Cleft palate (CP) is one of the most common birth defects in humanbeings. CP was still a great challenge to oral medicine，particularly for its pathogenesis.In general, CP was considered as a polygenic disease, and it can be caused by bothenvironmental and genetic factors. According to current report, tongue became flat anddecline, after that the lateral palate take a rotation and extend toward the horizontal line,contact and fuse at Embryo day14. Therefore, the difference or defect of the tonguedevelopment was one of the mechanisms of cleft palate. However, what cause thesedifferences was not clear. We studied the expression profile of whole genome fortongue tissue in CP, which will help better understand the regulation mechanisms oftongue development on molecular level, and biological function of significant genes inboth health and CP. Thus provide a new insight to both dig molecular target fordiagnosing CP and explore effective methods for treating CP.Gene chip technology, also known as DNA microarray. it can analysis thousandsof DNA fragments at the same time, a large number of probe molecules is fixed on thesupport material with the labeled samples molecular hybridization, the hybridizationsignal intensity of each detection probe molecule and then get the number and sequenceinformation of the sample molecules. At the same time, and has broad potentialapplication value in the detection, prevention of human disease and it play an importantrole in the life sciences.Objective: Screening differentially expressed genes of retinoic acid cleft palatetongue tissue in mice by using gene chips technology and analysis the function and itsassociated signaling pathway, observed changes and possible biological significance ofdifferences gene in normal development and cleft palate formation.Methods: We gave all-trans RA (100mg/kg) to E10ICR pregnant mice andinduce animal model with cleft palate.After that obtain embryos of13.5days in the experimental group and control group of mice tongue tissues to do Affymetrix Mouse4302.0expression profiling chip.And Preliminary analysis of expression genes thencombined with the GO classification to analyze function of differences genes. Screenfurther analysis of signal transduction signaling pathway from BioCarta and keggdatabase and further analysis of signal transduction related genes by literature inthe process of development of cleft palate.Results:1. In the basic of gene chips, Using the2-foldchange cut off, weidentified differentially expressed genes2146,1457up-regulated transcripts and689down-regulated ones, has not been annotated631,1515has been annotated.2. Go classification on the basis of the biological processes showed that certainfunctional categories of genes were over-represented such as glycosaminoglycanbiosynthetic process, cerebellum development, positive regulation of mesenchymal cellproliferation, actin polymerization or depolymerization. low-represented such as neuraltube closure, Wnt receptor signaling pathway, extracellular matrix organization, positiveregulation of cell proliferation.3. From the perspective of the signaling pathway, we have major differences genecorresponding signal transduction pathway such as Metabolic pathways,Wnt SignalingPathway, Hedgehog Pathway, MAPK signaling pathway and so on, they may play animportant role in the development of cleft palate.Conclusions: In this study, we found thousands of differentially expressed geneswhich related to multiple signaling pathways in the retinoic acid-induced cleft palatemice of tongue tissue in gene chip technology and we discuss preliminary their function,and prompted a tongue developmental abnormalities may be a direct result of cleftpalate.