The aim of this study was to test if mRNA encoding NKCC1, NKCC2, and Na⁺-coupled acid/base transporters and the corresponding proteins are expressed in rodent sweat glands, and, if expressed, determine the cellular and subcellular localization in rat, mouse, and human eccrine sweat glands. NKCC1 mRNA was demonstrated in rat palmar tissue, i.e. including sweat glands, by RT-PCR, whereas NKCC2 mRNA was absent. Also, NHE1 mRNA was demonstrated in rat palmar tissue whereas NHE2, NHE3, NHE4, NBCe1, NBCe2, NBCn1 and NCBE mRNA was not detected. The expression of NKCC1 and NHE1 proteins was confirmed in rat palmar skin by immunoblotting, whereas NKCC2, NHE2, and NHE3 proteins were not detected. Immunohistochemistry was performed using sections from rat, mouse, and human palmar tissue. Immunoperoxidase labeling revealed abundant expression of NKCC1 and NHE1 in the basolateral domain of secretory coils of rat, mouse, and human sweat glands and low expression in the coiled part of the ducts. In contrast, NKCC1 and NHE1 labeling was absent from rat, mouse, and human epidermis. Immunoelectron microscopy demonstrated abundant NKCC1 and NHE1 labeling of the basolateral plasma membranes of mouse sweat glands with no labeling of the apical plasma membranes or intracellular structures. The basolateral NKCC1 of the secretory coils of sweat glands would most likely account for the observed bumetanide sensitive NaCl secretion in the secretory coils and the basolateral NHE1 is likely to be involved in sodium-coupled acid-base transport