Characterization of plasma membrane Na+/H+ exchange in eel (Anguilla anguilla) intestinal epithelial cells.
The ability of eel intestinal epithelial cells to recover from an acute acid load was analysed using the fluorescent dye 2',7'-bis-carboxy-ethyl-5,6-carboxyfluorescein (BCECF) and cell suspensions. Under these experimental conditions (bicarbonate-free solutions) the resting pHi in cells prepared from sea-water (7.52 +/- 0.031) and fresh-water (7.50 +/- 0.094) adapted animals proved to be similar. The recovery rate (following an acid load) increases by increasing the Na ion concentration in the extracellular medium. This pHi recovery is competitively inhibited by the specific inhibitor dimethylamiloride (DMA) with a low Ki in sea- (1.2 microM) as well as in fresh-water (1.3 microM) adapted animals, indicating the presence of a specific Na/H exchange activity in these cells. Using basolateral membrane vesicles it could be demonstrated that this activity is located on the basolateral side of the enterocyte membrane. The kinetic parameters (Kapp and Jmax) of this exchanger are similar in fresh-water and sea-water adapted animals suggesting that no salinity adaptation occurs, thus excluding the involvement of the antiporter in the osmoregulatory processes. These results are in agreement with the presence in the plasma membrane of the eel enterocytes of a Na/H-1 (housekeeper) form of the antiporter.
Vilella
S
S
Dipartimento di Biologia, Università di Lecce, Italy.
Zonno
V
V
Lapadula
M
M
Verri
T
T
Storelli
C
C
D000584:Amiloride / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D002021:Buffers; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D004524:Eels / Q000378:metabolism*; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*
D002021:Buffers; D017923:Sodium-Hydrogen Exchangers; C039465:5-dimethylamiloride; D000584:Amiloride; D012964:Sodium; D011188:Potassium