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Immunofluorescence was used to examine the expression and the cellular and subcellular localization of five subunits of the V-ATPase and some of their respective isoforms in rat epididymis and vas deferens: A, C1, C2, G1, G3, a1, a2, a4, d1, and d2. All five subunits were detected in epithelial cells of these organs but showed specific patterns of expression (Fig. 10). Narrow and Clear Cell Localization The V1 sector subunits A, C1, C2, G1, and G3 were all expressed in narrow and clear cells where they were located in their apical pole. These results are in agreement with the previously reported expression of other V1 subunits, B1, B2, and E2, in these cells and further confirm the high expression of the V-ATPase in acidifying clear cells. Our laboratory has shown that these cells exhibit a significant bafilomycin-dependent proton secretion and are responsible for the majority of proton secretion in the rat vas deferens. Net proton secretion in these cells is regulated via recycling of V-ATPase-containing vesicles between apical microvilli and intracellular vesicles.