""Tc-Pertechnetate ("'rcO) has widespread clinical use in the diagnosis and evaluation of dysfunctions in many different tissues. However, despite the broad clinical application of this radionuclide, very little is known about the mechanism by which ""TcOi enters a cell. We report evidence here that ""'TcO4shares the Na+/K+/Cl co-transport system localized to the basolateral membrane of rat parotid acinar cells. " TcO. uptake by these cells was quite rapid (t1/2 30 s), was completely inhibited by the loop diuretics furosemide and bumetanide, and was markedly dependent on the presence of Na', K+, and Oin the extracellular medium. Relative to uptake measured in the presence of physiological extracellular salt concentrations (Hanks' salts), "'TcOj uptake was inhibited 80% by sodium replacement and 50% by potassium replacement. When Cl-was replaced with the physiologically inert anion gluconate a threefold stimulation in ""'TcO uptake resulted. These observations provide strong evidence that "'TcO4 can substitute for G-as a substrate for the Na+/K+/Cl-co-transporter and indicate that "'TcO4 uptake by salivary glands (e.g., as seen with salivary scintiscans), and possibly by a variety of other tissues, reflects the functional activity of this co-transport mechanism.