| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Research Abstract |
(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
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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
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