Epithelial sodium channels (ENaC) are uniformly distributed on motile cilia in the oviduct and the respiratory airways
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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.
KeywordsExtracellular fluid Immunohistochemistry Ion channels Lung Pseudohypoaldosteronism Tubulin Axoneme CFTR
This research was funded in part by a grant from the Chief Scientist of the Israel Ministry of Health. The authors thank Mr. Tevie Mehlman (Biological Mass Spectrometry Unit, Weizmann Institute of Science) for assistance with mass spectrometry analysis. This work was carried out in partial fulfillment of the requirements for a Ph.D. degree of Y.E. at the Sackler Faculty of Medicine of Tel Aviv University.
- Chang SS, Grunder S, Hanukoglu A, Rosler A, Mathew PM, Hanukoglu I, Schild L, Lu Y, Shimkets RA, Nelson-Williams C, Rossier BC, Lifton RP (1996) Mutations in subunits of the epithelial sodium channel cause salt wasting with hyperkalaemic acidosis, pseudohypoaldosteronism type 1. Nat Genet 12:248–253PubMedCrossRefGoogle Scholar
- Duc C, Farman N, Canessa CM, Bonvalet JP, Rossier BC (1994) Cell-specific expression of epithelial sodium channel alpha, beta, and gamma subunits in aldosterone-responsive epithelia from the rat: localization by in situ hybridization and immunocytochemistry. J Cell Biol 127:1907–1921PubMedCrossRefGoogle Scholar
- Lange K (2011) Fundamental role of microvilli in the main functions of differentiated cells: outline of an universal regulating and signaling system at the cell periphery. J Cell Physiol 226: 896–927Google Scholar
- Williams CL, Li C, Kida K, Inglis PN, Mohan S, Semenec L, Bialas NJ, Stupay RM, Chen N, Blacque OE, Yoder BK, Leroux MR (2011) MKS and NPHP modules cooperate to establish basal body/transition zone membrane associations and ciliary gate function during ciliogenesis. J Cell Biol 192:1023–1041PubMedCrossRefGoogle Scholar