Developmental maxillofacial morphogenesis and relevant molecular regulatory mechanisms in mice

本研究旨在阐明小鼠下颌突融合和腭发育的分子调控。使用ICR小鼠胚胎。从E9.5-11.5的咽弓和下颌弓的内侧区域以及E13.5-14.5的腭突获得用于基因表达和组织形态学的样本。本研究从组织学和免疫组化的角度探讨了小鼠颅面形态发生的多方面事件和表型特征。基因表达谱分析芯片和实时荧光PCR相结合。我们还探讨了网站离散的基因表达谱，通过比较下颌突的中间和侧面的部分。使用激光捕获显微切割从冷冻切片中收集指定内侧区域的间充质细胞群。cDNA微阵列分析揭示了转录因子的不同时空表达模式（例如，Pax，Runx，Snaih，一束生长因子和受体（如 ...更多信息 例如，Shh、FGF、TGF β、BMR ET 1、）、Rho家族和细胞粘附分子（N-钙粘蛋白、CD 44）。感兴趣的是，大量的基因（即，Ptx 1、Dlx、dHand、eHand、Msx 1、Bapx 1、Barx 1、Fgf 2、Fgf 8、ET 1）被认为参与了上皮-间充质相互作用，被鉴定为在融合过程中在下颌弓内侧区域差异表达的候选者。与骨骼肌分化相关的基因也被验证为上调或下调，根据完成内侧融合的下颌突和随后的舌头的发展。在平行的研究中，我们开发了器官培养模型，以吸收在下颌骨融合和舌形态发生中涉及的分子和细胞动力学的体内事件。在基金期，我们验证了SHH和FGF信号对下颌突融合和肌源性细胞增殖、凋亡和分化的影响。包括运动中发育小鼠颅面结构的3D构造的结果在网站（http：//www.ndu.ac.jp/pathhome/）以及出版物和摘要的形式中呈现。少

The present study aimed at elucidating the molecular regulation of mandibular process-fusion and palatogenesis in mouse development. ICR mouse embryos were used. Specimens for gene expression and histomorphology were obtained from the medial region of the pharyngeal and mandibular arches at E9.5-11.5 and palatal processes at E13.5-14.5. Multifaceted events and phenotypic features of mouse craniofacial morphogenesis were investigated histologically and immunohistochemically. Gene expression profiles were analyzed in combination of microarray and real-time PCR. We also explored site-discrete gene expression profiles by comparing the median and lateral portions of the mandibular processes. Mesenchymal cell populations in the specified medial region were collected from cryosections using laser capture microdissection. cDNA microarray analysis disclosed distinct spatial-temporal expression patterns of transcription factors (e.g., Pax, Runx, Snaih, a bundle of growth factors and receptors (e … More .g., Shh, FGFs, TGF β, BMR ET1,), Rho families, and cell adhesion molecules (N-cadherin, CD44). Of interest, a large number of the genes (i.e., Ptx1, Dlx, dHand, eHand, Msx1, Bapx1, Barx1, Fgf2, Fgf8, ET1), which are supposed to be involved in epithelio-mesenchymal interaction, were identified as candidates that were differentially expressed at the medial region of mandibular arches during the fusion process. Genes relevant to skeletal muscle differentiation were also validated to be up- or down-regulated in accordance with the completion to medial fusion of mandibular processes and subsequent tongue development. In parallel studies, we developed organ culture models to assimilate in vivo events of molecular and cellular dynamics involved in mandibular fusion and tongue morphogenesis. In the funded period, we verified the effects of SHH and FGF signals on mandibular process fusions and myogenic cell proliferation, apoptosis and differentiation. The results including 3D constructs of developing mouse craniofacial structures in motion are presented in Web site (http://www.ndu.ac.jp/~pathhome/) as well as in forms of publications and abstracts. Less