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To investigate the biology of the male genital duct epithelium, we have established cell cultures from the ovine vas deferens and epididymis epithelium. These cells develop tight junctions, high transepithelial electrical resistance, and a lumen-negative transepithelial potential difference as a sign of active transepithelial ion transport. In epididymis cultures the equivalent short-circuit current (I(sc)) averaged 20.8+/-0.7 microA/cm(2) (n = 150) and was partially inhibited by apical application of amiloride with an inhibitor concentration of 0.64 microM. In vas deferens cultures, I(sc) averaged 14.4+/-1.1 microA/cm(2) (n = 18) and was also inhibited by apical application of amiloride with a half-maximal inhibitor concentration (K(i)) of 0.68 microM. The remaining amiloride-insensitive I(sc) component in epididymis and vas deferens cells was partially inhibited by apical application of the Cl(-) channel blocker diphenylamine-2-carboxylic acid (1 mM). It was largely dependent on extracellular Cl(-) and, to a lesser extent, on extracellular HCO(-)(3). It was further stimulated by basolateral application of forskolin (10(-5) M), which increased I(sc) by 3.1+/-0.3 microA/cm(2) (n = 65) in epididymis and 0.9+/-0.1 microA/cm(2) (n = 11) in vas deferens. These findings suggest that cultured ovine vas deferens and epididymis cells absorb Na(+) via amiloride-sensitive epithelial Na(+) channels (ENaC) and secrete Cl(-) and HCO(-)(3) via apical cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channels. This interpretation is supported by RT-PCR data showing that vas deferens and epididymis cells express CFTR and ENaC mRNA.

Type

Journal article

Journal

American journal of physiology. Cell physiology

Publication Date

05/2000

Volume

278

Pages

C885 - C894

Addresses

University Laboratory of Physiology, Oxford OX1 3PT, United Kingdom.

Keywords

Epididymis, Vas Deferens, Cells, Cultured, Animals, Sheep, Anthranilic Acids, Forskolin, Amiloride, Cystic Fibrosis Transmembrane Conductance Regulator, Sodium Channels, RNA, Messenger, DNA Primers, Microscopy, Electron, Electrophysiology, Cell Differentiation, Gene Expression, Ion Transport, Base Sequence, Male, Epithelial Sodium Channel