H-K-ATPase immunoreactivity in cortical and outer medullary collecting duct

Abstract

Biochemical and physiologic studies in individual segments of the cortical collecting duct (CCD) and outer medullary collecting duct (OMCD) have provided evidence for the presence of an H-K-ATPase which is involved in the reabsorption of potassium in exchange for protons. The present study was designed to determine the cellular distribution of H-K-ATPase immunoreactivity in the CCD and OMCD of the rat and rabbit using mouse monoclonal antibodies against hog gastric H-K-ATPase. Kidneys of normal rats and rabbits were preserved for light microscopic immunohistochemistry and embedded in paraffin. Sections were incubated with the primary antibody followed by the avidin-biotin-horseradish peroxidase procedure. Sections incubated without primary antibody or with a non-specific mouse Ig served as controls. Light microscopy revealed diffuse cytoplasmic staining indicating H-K-ATPase immunoreactivity in intercalated cells in the CCD and OMCD in both rat and rabbit. In all segments studied except the rat CCD, the percentage of H-K-ATPase immunoreactive cells corresponded to the percentage of intercalated cells. In the rat CCD only 23% of the cells were reactive with H-K-ATPase antibodies, which is less than the percentage of intercalated cells in this region. It is possible that only type A intercalated cells possess H-K-ATPase immunoreactivity or that some intercalated cells did not have sufficient activity to be detected by our method. These results demonstrate H-K-ATPase immunoreactivity in the intercalated cells of the CCD and OMCD of rat and rabbit, suggesting that these cells are involved in potassium reabsorption in exchange for proton secretion in the mammalian collecting duct.