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*
Unit of Molecular Toxicology, Institute for Medical Research and
Occupational Health, Zagreb, Croatia
Renal Unit and Program in Membrane Biology, Massachusetts General Hospital
and Department of Pathology, Harvard Medical School, Boston,
Massachusetts.
Correspondence to Dr. Ivan Sabolic, Institute for Medical Research and Occupational Health, Ksaverska cesta 2, P.O. Box 291, 10001 Zagreb, Croatia. Phone: 385 1 4673 188; Fax: 385 1 4673 303; E-mail: sabolic{at}mimi.imi.hr
Abstract. The Na/K-ATPase plays a fundamental role in the
physiology of various mammalian cells. In the kidney, previous
immunocytochemical studies have localized this protein to the basolateral
membrane in different tubule segments. However, intercalated cells (IC) of the
collecting duct (CD) in rat and mouse were unlabeled with anti-Na/K-ATPase
antibodies. An antigen retrieval technique has been recently described in
which tissue sections are pretreated with sodium dodecyl sulfate before
immunostaining. This procedure was used to reexamine the presence of
Na/K-ATPase in IC along the rat nephron using monoclonal antibodies against
the Na/K-ATPase 
-subunit. Subtypes of IC along the nephron were
identified by their distinctive staining with polyclonal and monoclonal
antibodies to the 31-kD vacuolar H+-ATPase subunit, whereas
principal cells (PC) were labeled with a polyclonal antibody to the water
channel aquaporin-4 (AQP-4). In PC, the Na/K-ATPase and AQP-4 staining
colocalized basolaterally. In contrast to previous reports, we found that IC
of all types showed basolateral labeling with the anti-Na/K-ATPase antibody.
The staining was quantified by fluorescence image analysis. It was weak to
moderate in IC of cortical and outer medullary collecting ducts and most
intense in IC of the initial inner medullary collecting duct. IC in the
initial inner medulla showed a staining intensity that was equivalent or
stronger to that in adjacent principal cells. Models of ion transport at the
cellular and epithelial level in rat kidney, therefore, must take into account
the potential role of a basolateral Na/K-ATPase in intercalated cell
function.
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