Bicarbonate absorption in eel intestine: evidence for the presence of a membrane-bound carbonic anhydrase on the brush-border membranes of the enterocyte

icarbonate absorptive fluxes through the isolated intestine of the European eel (Anguilla anguilla) were evaluated by the pH-stat method under short-circuited conditions. It was found that bicarbonate absorptive flux was dependent on the luminal Na+ and was inhibited by luminal 4-acetamido-4' stilbene-2-2' disulfonic acid (SITS; 2.5 x 10(-4) M) and luminal acetazolamide (10(-4) M), while luminal amiloride (1 mM) was without effect. Furthermore, by using brush border membrane vesicles (BBMV) isolated from eel intestine, the existence of two carbonic anhydrase (CA) isoforms, one tightly associated to the brush border membrane (BBM) and the other soluble in the cytosol, was demonstrated. The membrane-bound CA differs from the cytoplasmic isoform in that 1) it is relatively resistant to treatment with 0.045% lauryl sulfate sodium salt (SDS); 2) it is less inhibitable by ethoxzolamide and sulfanilamide; and 3) its Km(app) is significantly lower than that of the cytoplasmic isoform. These results suggest that a BBM-bound CA isozyme would play an important role in bicarbonate absorption from the lumen, facilitating the HC0(3)(-) transfer through the luminal membrane of the eel enterocyte most likely via a Na+ (HCO3-) or (OH-) cotransport system.