Microscopical Methods for the Localization of Na+,K+-ATPase

Na+,K+-ATPase plays a central role in the ionic and osmotic homeostasis of cells and in the movements of electrolytes and water across epithelial boundaries. Microscopic localization of the enzyme is, therefore, of crucial importance in establishing the subcellular routes of electrolyte flow across structurally complex and functionally polarized epithelia. Recently developed approaches to the localization of Na+,K+-ATPase are reviewed. These methods rely on different properties of the enzyme and encompass cytochemical localization of the K+-dependent nitrophenylphosphatase component of the enzyme, autoradiographic localization of tritiated ouabain binding sites, and immunocytochemical localization of the holoenzyme and of its catalytic subunit. The rationales for each of these techniques are outlined as are the criteria that have been established to validate each method. The observed localization of NA+,K+-ATPase in various tissues is discussed, particularly as it relative to putative and hypothetical mechanisms that are currently thought to mediate reabsorptive and secretory electrolyte transport.