Elucidation of Cellular and Molecular Pathogenesis of Maxillo-Mandibular and Facial Malformation and Disorders : Use of Gene-Modified Mice

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Our long-term strategy is to elucidate pathogenesis of varieties of malformations taking place in the maxillo-facial region. To this objective, the present project aimed to inspect phenotypic expression (and alterations) and relevant genetic network operating temporo-spatially during mouse embryonic development. We specifically focused attention on: (1) palategenesis and cleft formation, which is most frequently encountered among various malformations in humans, (2) development and anomalies of Meckel's cartilage and mandible, (3) early development and hypoplasia of temporomandibular joint (TMJ), and (4) cellular events involved in tongue development and its hypoplasia in Endothelin1 (-I-) mice. Cultures using the Trowell organ culture system were conducted to monitor in vitro cellular events in palatogenesis and tongue morphogenesis. Specimens in culture were maintained in a humidified incubator at 37℃ under 5% C0_2 using DMEM/Fl2 (1:1) medium. In 2004, gene expression patterns in secondary palate and tongue in vivo and in vitro were surveyed using 80 probes for RT-PCR and real-time PCR. We also continued analysis of gene expression in specific cell types, namely, snail and Rho family in medial edge epithelial cells during palategenesis and myogenic mater genes (Pax 3, MyoD, myogenin) in tongue primordium, which were isolated by laser-capture microdissection. Transgenic mice used were homozygous (-I-) for TGF-β, δEF1 or endothelin1, as well as Klotho (-I-) used for investigation of TMJ aging. Phenotypic features of these transgenic mice were scrutinized by means of histology and x-ray μCT in combination of 3D-reconstruction techniques. Imaging data collected during the funded period have been accumulated and tabulated in our database with the aid of a computer server, which was purchased in the last year. We at present are in the process to display them on the Web-site (http://www.ndu.ac.jp/~pathhome).