Expression And Localization Of VEGFs And Its Receptors In Morphogenesis Of Tooth Germs In Mouse

OBJECTIVE: The aim of this study is to preliminary analyze the gene expression as well as protein localization of VEGFs and VEGFRs in in morphogenesis of tooth germs and explore the possible role that VEGFs and VEGFRs played during tooth development.METHODS:(1) Mice at different gestational stages(E11.5, E13.5, E15.5, E16.5,E18.5 and PN1)were selected. The m RNA of the mandible(E11.5, E13.5) and tooth germ(E15.5, E16.5, E18.5, and PN1) was extracted and trancripted into c DNA. Expressions of VEGFs and VEGFRs m RNA in the mandible(E11.5, E13.5) and tooth germ(E15.5, E16.5, E18.5, and PN1) were measured by RT-PCR.(2) Localization of VEGFs and VEGFRs protein expression in the morphogenesis of tooth germ at E12,E14,E15,E16,E18 and PN1) were observed by immunefluorescence staining.RESULTS:(1) The c DNA library of tooth germ derived from different stages has been established through this research. It will provide a resource for accessing the expression of other genes in BALB/c mouse tooth germ. VEGFs and VEGFRs m RNA level in mandible seemed to increase while in tooth germ gradually decrease during tooth germ morphogenesis.(2) VEGFR-2 protein expression showed weak fluorescence in tooth germ. Strong fluorescence of VEGF-B, D, VEGFR-2,-3 was found in mandible cartilage, etc. VEGF-A, C and VEGFR-1protein expression barely showed fluorescence in tooth germ or in other maxillofacial organs.CONCLUSIONS: In this study, reverse transcription-polymerase chain reaction and immunofluorescence techniques have been combined to preliminary analyze the expression of VEGFs and VEGFRs m RNA in morphogenesis of in mouse molar germs. The expression of VEGF-B, VEGF-D, VEGFR-2(Flk-1) and VEGFR-3(Flt-4) protein has been localized in maxillofacial organs other than tooth germ. The result of this study is not adequate explain clearly the role of VEGFs and VEGFRs protein have played during development of tooth germ and other maxillofacial organs. It still needs to add both quantitative and qualitative analysis of proteins and nucleic acids, organ culture if it's necessary, into the study method to continue the study.