Effects of estrogen on proton secretion via the apical membrane in vaginal-ectocervical epithelial cells of postmenopausal women.
OBJECTIVE
To understand estrogen regulation of proton (H(+)) secretion by human vaginal-ectocervical epithelial cells and the mechanisms involved.


METHODS
Primary-tertiary cultures of human normal vaginal-ectocervical epithelial cells were generated from surgical specimens of premenopausal women (aged 37-46 years) and of postmenopausal women (aged 53-65 years). Cells were grown on filters, and measurements were made of changes in extracellular pH (pHo) in the contraluminal (CL) and luminal (L) solutions 30 minutes after shifting cells to basic salt solution.


RESULTS
Upon shifting cells to basic salt solution, CL-pHo decreased from 7.4 to 7.25, and was not affected by removal of intracellular estrogens or treatment with estradiol. L-pHo decreased from 7.4 to 7.05 in cells of premenopausal women, and from 7.4 to 7.20 in cells of postmenopausal women. Removal of intracellular estrogens attenuated the decrease in L-pHo in cells of premenopausal women (only to 7.20). In cells of premenopausal women stripped of estrogens, treatment with 10 nM 17beta-estradiol restored the decrease in L-pHo. In estrogen-stripped cells of postmenopausal women, treatment with estradiol augmented luminal acidification but to a lesser degree than in cells of premenopausal women (L-pHo of 7.15 vs 7.05). In cells of pre- and postmenopausal women, the addition in the L solution of bafilomycin-A(1), a specific inhibitor of the vacuolar-H(+)-ATPase (V-H(+)-ATPase), blocked the decrease in L-pHo.


CONCLUSIONS
Human vaginal-ectocervical epithelial cells acidify constitutively their luminal solution, and the effect is mediated by active H(+) secretion by V-H(+)-ATPase expressed predominantly in the apical cell membrane. Estrogen deprivation attenuates, and treatment with 17beta-estradiol augments, active H(+) secretion. Finally, cells of postmenopausal women actively secrete H(+) via apically located V-H(+)-ATPase, but the effect is lesser, and estrogen failed to augment active H(+) secretion, as in cells of premenopausal women. These data suggest that in addition to hypoestrogenism, other factors of the aging process affect the capacity of vaginal-ectocervical cells to secrete acid.
Gorodeski
George I
GI
Department of Reproductive Biology, CASE, Case Western Reserve University, Cleveland, OH, USA. gig@cwru.edu
D000143:Acids / Q000378:metabolism; D000328:Adult; D000368:Aged; D002584:Cervix Uteri / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism*; D004967:Estrogens / Q000494:pharmacology*; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D008875:Middle Aged; D017698:Postmenopause / Q000378:metabolism; D017697:Premenopause / Q000378:metabolism; D006180:Proton-Translocating ATPases / Q000378:metabolism; D011522:Protons; D014617:Vacuoles / Q000201:enzymology; D014621:Vagina / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
D000143:Acids; D004967:Estrogens; D011522:Protons; D006180:Proton-Translocating ATPases