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1 Program in Membrane Biology, Massachusetts General Hospital, Charlestown, Massachusetts 02129; 2 Groupe de Recherche en Transport Membranaire, University of Montreal, Montreal, Canada H3C3J7; 3 Department of Cellular and Molecular Physiology, Yale University, New Haven, Connecticut 06510; and 4 Department of Medicine, Harvard Medical School, Boston, Massachusetts 02215
An acidic luminal pH in the epididymis and vas deferens (VD) helps maintain mature sperm in an immotile state during storage. We have previously shown that the majority of proton secretion in the VD is due to the activity of the vacuolar H+-ATPase. Acidification is dependent on luminal sodium in more proximal regions of the epididymis, and we examined the distribution of the Na+/H+ exchanger, NHE3, by immunofluorescence and measured Na+/H+ exchange (NHE) activity in isolated epididymal tubules. NHE3 was detected in the apical pole of nonciliated cells of the efferent ducts and principal cells (PC) of the epididymis. No staining was seen in the distal cauda epididymidis and the VD. Isolated tubules from the distal initial segment (DIS) and proximal cauda epididymidis were perfused in vitro and loaded with the pH-sensitive dye 2',7'-bis(carboxyethyl)-5(6')-carboxyfluorescein. Ethylisopropyl amiloride (EIPA) (50 µM) reduced the initial rate of intracellular pH recovery (dpHi/dt), in response to an acute acid load, by 51% and 45% in the DIS and cauda epididymidis, respectively. In the DIS, removal of luminal sodium reduced dpHi/dt by 52%. HOE694 (50 µM) inhibited all EIPA-sensitive dpHi/dt in the DIS, despite the previously reported absence of NHE2 in this region (Cheng Chew SB, Leung GPH, Leung PY, Tse CM, and Wong PYD, Biol Reprod 62: 755-758, 2000). These data indicate that HOE694- and EIPA-sensitive Na+/H+ exchange may participate, together with the H+-ATPase, in luminal acidification in the male excurrent duct.
male reproductive tract; transepithelial acid-base transport; acidification; immunofluorescence; intracellular pH.
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