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/HCO3
exchangers of rat thick limbs
1 Laboratoire de Physiologie et Endocrinologie Cellulaire Rénale, Faculté de Médecine Broussais-Hôtel Dieu, Institut National de la Santé et de la Recherche Médicale, Unité 356, Paris, France; and the 2 Departments of Cell Biology and Medicine, Harvard Medical School, and the Molecular Medicine and Renal Units, Beth Israel Hospital, Boston, Massachusetts 02215
Cl
/HCO3
exchange was measured in luminal (LMV) and basolateral
(BLMV) membrane vesicles purified from rat medullary thick ascending
limb (MTAL).
Cl/HCO3
exchange in BLMV and LMV was inhibited by DIDS, with respective
IC50 values of 3.2 ± 0.9 and 15.2 ± 5.2 µM, whereas
Cl conductances were DIDS
insensitive. At constant external pH, BLMV
36Cl/HCO3
and
36Cl/Cl
exchanges exhibited a sigmoidal pattern of activation as internal pH
(pHi) increased from 6.1 to 8.0, whereas LMV
36Cl/Cl
exchange was unchanged between pHi
6.7 and 7.8. The 165-kDa AE2 polypeptide and ~115-kDa AE1-related
polypeptide were present only in BLMV. In contrast, AE1-related
polypeptides of ~90 and 95 kDa were present not only in BLMV but also
(in variable abundance) in LMV. We conclude that rat MTAL BLMV and LMV
express distinct anion exchange activities and distinct sets of AE
polypeptides. AE2 (and perhaps AE1) in BLMV likely contribute to
HCO3 absorption. In contrast, LMV
exchangers may contribute to NaCl absorption via parallel coupling with
the luminal
Na+/H+
antiporters and/or may provide negative feedback regulation of HCO3 absorption.
anion exchange; AE2; band 3; AE1; 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid; membrane vesicles; immunoblot
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