Axonemal localization of the dynein component DNAH5 is not altered in secondary ciliary dyskinesia
Axonemal localization of the dynein component DNAH5 is not altered in secondary ciliary dyskinesia
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DOI：
10.1203/01.pdr.0000200809.21364.e2
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发表时间：
2006-03-01
影响因子：
3.6
通讯作者：
Omran, H
中科院分区：
文献类型：
--
作者：
Olbrich, H;Horváth, J;Omran, H
原发性纤毛运动障碍（PCD）是一种异质性遗传性疾病，其特征是半数受影响个体的反复呼吸道感染和内脏逆位。目前的诊断依赖于纤毛超微结构异常或纤毛搏动改变。炎症引起的继发性纤毛运动障碍（SCD）的改变往往使诊断工作复杂化。我们最近发现，在呼吸道上皮细胞的PCD患者与外部动力蛋白臂缺陷的动力蛋白DNAH 5是错误定位和完全或部分缺席的纤毛轴丝。在这项研究中，我们解决了SCD是否可能影响轴丝DNAH 5定位在呼吸细胞的问题。为了在体外诱导SCD，我们用白细胞介素-13（IL-13）处理原代人呼吸道上皮细胞培养物。分别用高速摄像显微镜和透射电镜观察纤毛功能和超微结构。对于DNAH 5的体内定位，我们对有SCD证据的患者进行了鼻腔刷检活检。通过免疫荧光显微镜分析DNAH 5的表达。IL-13处理的细胞显示SCD的证据。纤毛搏动频率显着降低，超微结构分析显示轴丝解体与对照细胞相比。高分辨率免疫荧光研究呼吸道上皮细胞与SCD确定在体外和体内正常轴丝DNAH 5定位。DNAH 5定位不改变SCD，表明免疫荧光分析作为一种新的诊断工具，在PCD的高潜力。
Primary ciliary dyskinesia (PCD) is a heterogeneous genetic disorder characterized by recurrent airway infections and situs inversus in half of affected individuals. Diagnosis currently relies on demonstration of abnormal ciliary ultrastructure or altered ciliary beat. Alterations encountered in secondary ciliary dyskinesia (SCD) caused by inflammation often complicate the diagnostic workup. We have recently shown that in respiratory epithelial cells from PCD patients with outer dynein arm defects the dynein protein DNAH5 is mislocalized and either completely or partially absent from the ciliary axoneme. In this study, we addressed the question whether SCD might affect axonemal DNAH5 localization in respiratory cells. To induce SCD in vitro, we treated primary human respiratory epithelial cell cultures with interleukin-13 (IL-13). Ciliary function and ultrastructure were assessed by high-speed videomicroscopy and transmission electron microscopy, respectively. For in vivo localization of DNAH5, we performed nasal brushing biopsies in patients with evidence of SCD. Expression of DNAH5 was analyzed by immunofluorescence microscopy. IL-13-treated cells showed evidence of SCD. Ciliary beat frequency was significantly reduced and ultrastructural analyses showed axonemal disorganization compared with control cells. High-resolution immunofluorescence studies of respiratory epithelial cells with SCD identified in vitro and in vivo normal axonemal DNAH5 localization. DNAH5 localization is not altered by SCD, indicating a high potential for immunofluorescence analysis as a novel diagnostic tool in PCD.