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首页> 外文期刊>American Journal of Physiology >Isolation and characterization of cholangiocyte primary cilia.
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Isolation and characterization of cholangiocyte primary cilia.

机译：胆管原发性纤毛的分离和鉴定。

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摘要

Primary cilia are distinct organelles expressed by many vertebrate cells, including cholangiocytes; however, their functions remain obscure. To begin to explore the physiological role of these organelles in the liver, we described the morphology and structure of cholangiocyte cilia and developed new approaches for their isolation. Primary cilia were present only in bile ducts and were not observed in hepatocytes or in hepatic arterial or portal venous endothelial cells. Each cholangiocyte possesses a single cilium that extends from the apical membrane into the bile duct lumen. In addition, the length of the cilia was proportional to the bile duct diameter. We reproducibly isolated enriched fractions of cilia from normal rat and mouse cholangiocytes by two different approaches as assessed by scanning electron, transmission electron, and confocal microscopy. The purity of isolated ciliary fractions was further analyzed by Western blot analysis using acetylated tubulin as a ciliary marker and P2Y(2) as a nonciliary cell membrane marker. These novel techniques produced enriched ciliary fractions of sufficient purity and quantity for light and electron microscopy and for biochemical analyses. They will permit further assessment of the role of primary cilia in normal and pathological conditions.
机译：原发纤毛是由许多脊椎动物细胞（包括胆管细胞）表达的截然不同的细胞器。但是，它们的功能仍然不清楚。为了开始探索这些细胞器在肝脏中的生理作用，我们描述了胆管细胞纤毛的形态和结构，并开发了分离它们的新方法。原发纤毛仅存在于胆管中，在肝细胞或肝动脉或门静脉内皮细胞中未观察到。每个胆管细胞都具有单个纤毛，纤毛从顶膜延伸到胆管腔。另外，纤毛的长度与胆管直径成正比。我们通过两种不同的方法，通过扫描电子，透射电子和共聚焦显微镜评估，从正常大鼠和小鼠胆管细胞中可再现地分离出纤毛富集部分。通过使用乙酰化微管蛋白作为睫状标记物和P2Y（2）作为非睫状细胞膜标记物的蛋白质印迹分析进一步分析分离的睫状级分的纯度。这些新技术产生了足够纯度和数量的富集的睫毛级分，用于光学和电子显微镜以及生化分析。他们将允许进一​​步评估原发性纤毛在正常和病理条件下的作用。

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