Molecular Biological Investigation of the mechanism of ciliogenesis

(1)关于纤毛根的分子组成知之甚少。我们提出了单克隆抗体的粗制备的纹状根从人输卵管分离，并获得了克隆（R4109），特异性标记的睫状根。与分泌细胞和成纤维细胞中的孤纤毛相关的小根也被标记。R4109通过免疫印迹鉴定了一个195 kDa的蛋白。免疫荧光显微镜下，R4109可标记幼鼠输卵管上皮中的纤毛细胞呈球状和/或颗粒状。免疫电镜显示它们分别对应于纤维颗粒复合体和致密颗粒。结果表明，195-kDa蛋白质是纹状根和致密颗粒共同的组分，从而表明致密颗粒参与了纹状根的形成。(2)我们报告一位13岁的女性，有咳嗽、痰和发烧。患者同时患有慢性鼻窦炎和支气管炎。胸部 ...更多信息 胸部X线和计算机断层扫描显示双侧下叶、右中叶和左上叶支气管粘液性支气管充盈和支气管扩张。根据这些发现，建议原发性睫状体运动障碍。这一点通过鼻活检和透射电子显微镜检查的结果得到证实，在鼻活检中，所有的微管在纤毛基体附近都有节段性缺陷。根据这些发现，我们诊断为原发性睫状体运动障碍，这可能是由于，至少部分，睫状体微管节段性缺陷的患者。(3)人输卵管上皮为单层柱状，由纤毛细胞和分泌细胞组成。孤立纤毛通常从分泌细胞的顶面伸出。将大鼠纤毛细胞纹状根粗品注射到大鼠体内，成功地获得了6株单克隆抗体（R38，R67，R95，R149，R155，R213），它们以相同的染色模式标记纤毛根。抗体通过免疫印迹鉴定205-215 kDa蛋白质。与孤纤毛的条纹状小根形状各异;直，弯曲，分支，辐射，和循环后重建的序列图像，使用共聚焦激光扫描显微镜。本研究首次对细根的整体形态进行了鉴定。利用这些抗体，我们研究了内脏器官中横纹小根的发育。除了某些细胞类型，如成纤维细胞，内皮细胞和肌上皮细胞，各种组织中的各种上皮细胞在二倍体中心粒周围发育条纹状小根，而不形成孤立的纤毛。少

(1) Little is known about the molecular composition of the ciliary rootlet. We raised monoclonal antibodies to a crude preparation of striated rootlets isolated from the human oviduct, and obtained a clone (R4109) that specifically labeled the ciliary rootlets. Rootlets associated with the solitary cilium in secretory cells and fibroblasts were also labeled. R4109 identified a 195 kDa protein by immunoblotting. Ciliogenic cells in the oviduct epithelium of young mice were labeled in the globular and/or granular pattern by R4109 by immunofluorescence microscopy. Immunoelectron microscopy showed that they corresponded to fibrogranular complex and dense granule, respectively. The result demonstrated that the 195-kDa protein is a component common to the striated rootlet and dense granule, and thus suggested that dense granules are involved in the rootlet formation.(2) We reported a 13-year-old woman with cough, sputum and fever. The patient had both chronic sinusitis and bronchitis. Chest … More X-ray and computed tomographic scan of the chest revealed mucous bronchial filling and bronchiectasia in bronchi of bilateral lower lobes, right middle lobe and left upper lobe. On the basis of these findings, primary ciliary dyskinesia was suggested. This was confirmed by the findings from nasal biopsy with transmission electron microscopy where all of the microtubules were segmentally defected near the basal body in the cilia. On the basis of these findings, we diagnosed the patient with primary ciliary dyskinesia which may be due, at least in part, to segmental defect of ciliary microtubules.(3) The human oviduct epithelium is simple columnar and consists of ciliated and secretory cells. Solitary cilia usually extend from the apical surface of secretory cells. By injecting a crude preparation of striated rootlets from ciliated cells to rats, we successfully obtained 6 monoclonal antibodies (R38, R67, R95, R149, R155, R213) which labeled ciliary rootlets in the same staining pattern. The antibodies identified 205-215 kDa proteins by immunoblotting. The striated rootlets associated with solitary cilia were variously shaped ; straight, curved, branched, radiated, and looped after reconstruction of serial images using a confocal laser scanning microscope. The whole figures of the rootlets were firstly identified in the present study. Using the antibodies, we investigated the development of striated rootlets in internal organs. Except certain cell types such as fibroblasts, endothelial cells, and myoepithelial cells, various epithelial cells in a variety of tissues develop striated rootlets around diplosomal centrioles without formation of the solitary cilium. Less