Department of Molecular Biology, Ariel University Center, Ariel 40700, Israel.
Histochem Cell Biol. 2012 Mar;137(3):339-53. doi: 10.1007/s00418-011-0904-1. Epub 2011 Dec 30.
Epithelial sodium channels (ENaCs) are located on the apical surface of cells and funnel Na(+) ions from the lumen into the cell. ENaC function also regulates extracellular fluid volume as water flows across membranes accompanying Na(+) ions to maintain osmolarity. To examine the sites of expression and intracellular localization of ENaC, we generated polyclonal antibodies against the extracellular domain of human α-ENaC subunit that we expressed in E. coli. Three-dimensional (3D) confocal microscopy of immunofluorescence using these antibodies for the first time revealed that ENaCs are uniformly distributed on the ciliary surface in all epithelial cells with motile cilia lining the bronchus in human lung and female reproductive tract, all along the fimbrial end of the fallopian tube, the ampulla and rare cells in the uterine glands. Quantitative analysis indicated that cilia increase cell surface area >70-fold and the amount of ENaC on cilia is >1,000-fold higher than on non-ciliated cell surface. These findings indicate that ENaC functions as a regulator of the osmolarity of the periciliary fluid bathing the cilia. In contrast to ENaC, cystic fibrosis transmembrane conductance regulator (CFTR) that channels chloride ions from the cytoplasm to the lumen is located mainly on the apical side, but not on cilia. The cilial localization of ENaC requires reevaluation of the mechanisms of action of CFTR and other modulators of ENaC function. ENaC on motile cilia should be essential for diverse functions of motile cilia, such as germ cell transport, fertilization, implantation, clearance of respiratory airways and cell migration.
上皮钠通道 (ENaC) 位于细胞的顶膜表面,将腔内的钠离子引导入细胞。ENaC 的功能还调节细胞外液体积,因为水伴随钠离子穿过膜流动,以维持渗透压。为了研究 ENaC 的表达部位和细胞内定位,我们针对人α-ENaC 亚基的细胞外结构域在大肠杆菌中表达产生了多克隆抗体。首次使用这些抗体进行的三维(3D)共聚焦免疫荧光显微镜观察显示,ENaC 在所有具有纤毛的上皮细胞的纤毛表面均匀分布,纤毛排列在人肺和女性生殖道的支气管、输卵管的顶端、输卵管壶腹部和子宫腺中的罕见细胞上。定量分析表明,纤毛使细胞表面积增加了>70 倍,纤毛上的 ENaC 含量比非纤毛细胞表面高>1000 倍。这些发现表明 ENaC 作为纤毛周围液体渗透压的调节剂发挥作用。与将氯离子从细胞质引导至腔的囊性纤维化跨膜电导调节因子 (CFTR) 不同,CFTR 主要位于顶侧,而不是纤毛上。ENaC 在纤毛上的定位需要重新评估 CFTR 及其他 ENaC 功能调节剂的作用机制。纤毛上的 ENaC 对于纤毛的各种功能(如生殖细胞运输、受精、着床、呼吸道清除和细胞迁移)应该是必不可少的。