Dynamic Expression Of Signaling Molecules In Mouse Cleft Palate Induced By Retinoic Acid

Backgroud:Cleft palate(CP)is one of the most common malformation among live births,occurring with an incidence of 1.82%o in China。CP is an affectability disease,it is caused by genetic factors and interaction with environmental factors.Retinoic Acid(RA)is the derivative of vitamin A,it is generally used at chemotherapeutics,dermatopathic medicine and additives of cosmetics.RA is one of the common environmental factor.The gene homology of human and mice is more than 99%,and the development of mouse cleft palate is similar with human,thus using mouse as the model of cleft palate to study its mechanisms is a trend.At the process of orofacial development,many signaling pathways,such as transformation growth factor(TGF),Wnt,fibroblast growth factor(FGF),control the interaction between embryonic palatal mesenchyme(EPM)and medial edge epithelium (MEE).In current study, high-flux gene chip and mouse model of RA-induced cleft palate were used to screen Wnt and FGF signal molecules which related with RA-induced cleft palate.RT-PCR technique was performed to evaluate the expression of signal molecules,such as Wnt3, Wnt8a,Fgf9 and Fgf10.Objective:In order to screen the signaling molecules involved in palatogenesis and cleft palate,to study the dynamic expression level in the different stages of palate development and cleft palate formation.Methods: Mouse model of retinoic acid(RA)-induced cleft palate was set up,the palate of control and RA treated group was taken at embryo day(ED)14.5 for gene chip analysis,and the expression level of Wnt3,Wnt8a,Fgf9 and Fgf10 were detected at ED 13.5-ED15.5 in control and RA treated group by using semi-quantitive RT-PCR. Results:1.In the result of gene chip, compared with control, in RA-induced cleft palate group the expression of Wnt8a and Fgf9 was down-regulated.In converse,Wnt3 and Fgf10 was up-regulated.2.During all different stage of palatogenesis of wild type,the intense expression of Wnt3,Wnt8a, Fgf9 and Fgf10 was showed with a continuous dynamic pattern.3.Compared with control,the expression level of Wnt3, Wnt8a, Fgf9 and Fgf10 in RA-induced cleft palate group showed valuable difference(P<0.05).Conclusions:Wnt and FGF signaling pathway participate in the development of palate,and in the process of RA-induced cleft palate,RA pathway may interact with Wnt and FGF signaling pathway.