Open AccessImmunohistochemical Demonstration of Cl- Pump in Type A Intercalated Cells of Rat Kidney.
Author(s) -
Toshiaki Higashida,
Xun-Ting Zeng,
Mitsuyoshi Hara,
Koichiro Omori,
Sei Sasaki,
Michiaki Orikasa,
Fujio Shimizu,
Chiyoko Inagaki
Publication year - 2000
Publication title -
japanese journal of pharmacology/japanese journal of pharmacology
Language(s) - English
Resource type - Journals
eISSN - 1347-3506
pISSN - 0021-5198
DOI - 10.1254/jjp.83.233
Subject(s) - intercalated cell , immunohistochemistry , kidney , microbiology and biotechnology , aquaporin 3 , antibody , chemistry , aquaporin 2 , blot , aquaporin 1 , epithelial polarity , diaphragm pump , immunocytochemistry , cell type , cell membrane , membrane , aquaporin , biology , cell , endocrinology , biochemistry , water channel , materials science , immunology , gene , mechanical engineering , engineering , micropump , nanotechnology , inlet
In order to demonstrate the localization of an ethacrynic acid-sensitive Cl- pump in the rat kidney, immunohistochemical analysis was performed using an anti-Cl- pump antibody raised against rat brain Cl- pump protein with confocal laser scanning microscopy. The antibodies against Na+,K+-ATPase, aquaporin 2 and a type B intercalated cell marker, 43-kDa protein, were also used for comparison. Anti-Cl- pump antibody recognized a 51-kDa renal protein of the same size as that in the brain on Western blots. Cl- -pump-like immunoreactivity was observed on the basolateral membranes of 42+/-3% of cortical collecting duct (CCD) cells and of 38+/-1% of outer medullar collecting duct (OMCD) cells. Such immunoreactivity in CCD was sometimes co-localized with Na+,K+-ATPase, but in OMCD, the Cl- pump-like immunoreactivity co-existed with neither Na+,K+-ATPase, aquaporin 2 nor the type B intercalated cell marker 43-kDa protein. Thus, the Cl- pump was demonstrated to be localized on the basolateral membranes of type A intercalated cells of cortical and medullary collecting ducts in the rat kidney.