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1 Renal Unit and Program in Membrane Biology, Massachusetts General Hospital, Charlestown 02129; 2 Department of Medicine, Harvard Medical School, Boston 02215; and 3 Renal Division, Brigham and Women's Hospital, Boston, Massachusetts 02115
Epithelial cells of the epididymis and
vas deferens establish an optimum luminal environment in which
spermatozoa mature and are stored. This is achieved by active
transepithelial transport of various ions including Cl
and H+. We investigated the localization of three closely
related members of the ClC family, ClC-3, ClC-4, and ClC-5, in the
epididymis and vas deferens. RT-PCR using mRNA isolated by laser
capture microdissection (LCM)-detected ClC-3 and ClC-5 transcripts but did not detect any ClC-4-specific transcript. Western blot and immunofluorescence analysis demonstrated that ClC-3 and ClC-5 proteins
are present in all regions of the epididymis and in the vas deferens.
ClC-5 is expressed exclusively in H+-ATPase-rich cells
(narrow and clear cells). Confocal microscopy showed that ClC-5
partially colocalizes with the H+-ATPase in the subapical
pole of clear cells. ClC-3 is strongly expressed in the apical membrane
of principal cells of the caput epididymidis and the vas deferens and
is less abundant in principal cells of the body and cauda epididymidis.
These findings are consistent with a potential role for ClC-3 in
transepithelial chloride transport by principal cells and for ClC-5 in
the acidification of H+-ATPase-containing vesicles in
narrow and clear cells. ClC-5 might facilitate endosome trafficking in
the epididymis, as has been proposed in the kidney.
chloride channels, vacuolar H+-ATPase, principal cells, clear cells; laser capture microdissection
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