Molecular Biological Investigation of the mechanism of ciliogenesis

(1)对纤毛细根的分子组成知之甚少。我们从人的输卵管中分离出一种粗制的横纹细根，我们提出了一种克隆(R4109)，该克隆可以特异性地标记纤毛细根。与分泌细胞和成纤维细胞中的孤立纤毛相关的根也被标记。免疫印迹法鉴定R4109蛋白大小为195 kDa。免疫荧光显微镜下R4109标记幼年小鼠输卵管上皮细胞呈球形和/或颗粒状。免疫电子显微镜显示，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的蛋白质是横纹小根和致密颗粒的共同成分，提示致密颗粒参与了小根的形成。(2)我们报道了一例13岁的女性患者，以咳嗽、痰和发热为主诉。患者同时患有慢性鼻窦炎和支气管炎。胸部…胸部X线和计算机断层扫描显示双肺下叶、右中叶和左上叶的支气管粘液充盈和支气管扩张。在这些发现的基础上，提出了原发性睫状体运动障碍的建议。透射电子显微镜下鼻部活检的结果证实了这一点，所有的微管在纤毛的基底体附近呈节段性缺陷。根据这些发现，我们诊断为原发性睫状体运动障碍，这可能至少部分归因于纤毛微管的节段性缺陷。(3)人输卵管上皮为单柱状，由纤毛细胞和分泌细胞组成。单生纤毛通常从分泌细胞的顶面延伸。通过向大鼠注射纤毛细胞的粗品，我们成功地获得了6株以相同染色模式标记睫状根的单抗(R38、R67、R95、R149、R155、R213)。抗体经免疫印迹鉴定为205-215 kDa的蛋白。用激光共聚焦扫描显微镜重建连续图像后，与孤立纤毛相关的条纹根的形状多种多样；直的、弯曲的、分支的、辐射的和环状的。本研究首次对小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纹状根的发育情况。除成纤维细胞、内皮细胞和肌上皮细胞等某些细胞类型外，各种组织中的各种上皮细胞在双体中心粒周围发育有条纹的小根，而不形成孤立的纤毛。较少

(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