Basolateral localization of anion exchanger 2 (AE2) and actin in acid-secreting (parietal) cells of the human stomach.
Basolateral uptake of chloride by the HCl-secreting parietal cells of the gastric (oxyntic) glands is most likely mediated by a HCO3-/Cl- anion exchange mechanism. Circumstantial evidence indicates that in rodents the anion exchange proceeds through an anion exchanger 2(AE2)-like membrane protein. In the present study, we raised antibodies against a bacterial fusion protein expressing a approximately 26-kDa portion of the human AE2 sequence. These antibodies were used to identify and localize AE2 in the human stomach. Here we report that the mucosa of the human stomach expresses an approximately 160-kDa immunoreactive form of AE2 containing the AE2-specific exoplasmic domain (Z-loop) as identified by polymerase chain reaction. Immunostaining specific for AE2 was restricted to the basolateral membrane domain of parietal cells and was also detected in small epithelial cells localized in the glandular isthmus region. The latter cells most likely represent pre-parietal cells. Parietal cells were identified by simultaneous and sequential labelling with antibodies against the gastric H+,K(+)-ATPase and actin, respectively. Both actin and the H+,K(+)-ATPase were localized along the apical membrane of parietal cells and the membrane of their secretory intracellular canaliculi. In addition, actin was shown to be colocalized with AE2 along the basolateral cell surface. Discontinuous staining for AE2 coincided with infoldings of the basolateral plasma membrane labelled by the actin antibody. These observations indicate that AE2 might be placed at specialized (folded) microdomains of the basolateral cell surface by linkage to the actin-based cytoskeleton.
Jöns
T
T
Institute of Anatomy, University of Würzburg, Germany.
Warrings
B
B
Jöns
A
A
Drenckhahn
D
D
D000199:Actins / Q000032:analysis; D000818:Animals; D000906:Antibodies; D017920:Antiporters / Q000032:analysis*; D001483:Base Sequence; D005455:Fluorescent Antibody Technique; D006801:Humans; D008969:Molecular Sequence Data; D010295:Parietal Cells, Gastric / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011817:Rabbits; D051381:Rats; D013552:Swine
D000199:Actins; D000906:Antibodies; D017920:Antiporters