Hormonal and purinergic stimulation of bicarbonate secretion in oviducts of rhesus monkey

M. Rajagopal, H. Fischer, Jonathan Widdicombe

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Abstract

Because an increase in the HCO3 - concentration of oviductal liquid at midcycle is believed to markedly enhance fertility, we have studied active secretion of HCO3 - across highly differentiated cultures of monkey oviductal epithelium. Cultured cell sheets were mounted in Ussing chambers and bathed in medium containing 25 mM HCO 3 -. Purinergic agents potently stimulated short-circuit current (Isc) with an initial transient response declining within ∼2 min to a sustained response. The potency sequence of ATP ≈ UTP > ADP >> AMP suggested that the Isc response was mediated mainly by P2Y2 receptors. Acetazolamide, an inhibitor of carbonic anhydrase, had little or no effect on baseline Isc or the transient response to ATP but abolished the sustained response to ATP. Similar results were obtained on sheets of native epithelium. In pH-stat experiments, the abluminal medium of cell cultures was bathed in HCO3 --CO2 medium, and the pH of the unbuffered luminal medium was maintained at ∼7.4 by addition of strong acid or base. ATP stimulated base secretion, and this was inhibited by acetazolamide. Furthermore, these changes in secretion of base were in good quantitative agreement with the Isc responses. When phenol red (an estrogen) was removed from the culture medium, ATP-dependent HCO 3 - secretion was markedly reduced but could be restored by treatment with estradiol. Estrogens also markedly increased ciliation of the cultures. These results suggest that the midcycle increase in the HCO 3 - concentration of oviductal liquid may be mediated by the effects of estradiol on purinergic pathways or on ATP secretion.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Volume295
Issue number1
DOIs
StatePublished - Jul 2008

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Oviducts
Bicarbonates
Macaca mulatta
Adenosine Triphosphate
Acetazolamide
Cell culture
Transient analysis
Purinergic Agents
Estradiol
Estrogens
Epithelium
Purinergic P2Y2 Receptors
Phenolsulfonphthalein
Carbonic Anhydrase Inhibitors
Uridine Triphosphate
Liquids
Adenosine Monophosphate
Short circuit currents
Adenosine Diphosphate
Haplorhini

Keywords

  • Adenosine 5′-triphosphate
  • Carbonic anhydrase
  • Estradiol
  • Short-circuit current

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)
  • Endocrinology, Diabetes and Metabolism
  • Endocrinology
  • Biochemistry

Cite this

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title = "Hormonal and purinergic stimulation of bicarbonate secretion in oviducts of rhesus monkey",
abstract = "Because an increase in the HCO3 - concentration of oviductal liquid at midcycle is believed to markedly enhance fertility, we have studied active secretion of HCO3 - across highly differentiated cultures of monkey oviductal epithelium. Cultured cell sheets were mounted in Ussing chambers and bathed in medium containing 25 mM HCO 3 -. Purinergic agents potently stimulated short-circuit current (Isc) with an initial transient response declining within ∼2 min to a sustained response. The potency sequence of ATP ≈ UTP > ADP >> AMP suggested that the Isc response was mediated mainly by P2Y2 receptors. Acetazolamide, an inhibitor of carbonic anhydrase, had little or no effect on baseline Isc or the transient response to ATP but abolished the sustained response to ATP. Similar results were obtained on sheets of native epithelium. In pH-stat experiments, the abluminal medium of cell cultures was bathed in HCO3 --CO2 medium, and the pH of the unbuffered luminal medium was maintained at ∼7.4 by addition of strong acid or base. ATP stimulated base secretion, and this was inhibited by acetazolamide. Furthermore, these changes in secretion of base were in good quantitative agreement with the Isc responses. When phenol red (an estrogen) was removed from the culture medium, ATP-dependent HCO 3 - secretion was markedly reduced but could be restored by treatment with estradiol. Estrogens also markedly increased ciliation of the cultures. These results suggest that the midcycle increase in the HCO 3 - concentration of oviductal liquid may be mediated by the effects of estradiol on purinergic pathways or on ATP secretion.",
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T1 - Hormonal and purinergic stimulation of bicarbonate secretion in oviducts of rhesus monkey

AU - Rajagopal, M.

AU - Fischer, H.

AU - Widdicombe, Jonathan

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N2 - Because an increase in the HCO3 - concentration of oviductal liquid at midcycle is believed to markedly enhance fertility, we have studied active secretion of HCO3 - across highly differentiated cultures of monkey oviductal epithelium. Cultured cell sheets were mounted in Ussing chambers and bathed in medium containing 25 mM HCO 3 -. Purinergic agents potently stimulated short-circuit current (Isc) with an initial transient response declining within ∼2 min to a sustained response. The potency sequence of ATP ≈ UTP > ADP >> AMP suggested that the Isc response was mediated mainly by P2Y2 receptors. Acetazolamide, an inhibitor of carbonic anhydrase, had little or no effect on baseline Isc or the transient response to ATP but abolished the sustained response to ATP. Similar results were obtained on sheets of native epithelium. In pH-stat experiments, the abluminal medium of cell cultures was bathed in HCO3 --CO2 medium, and the pH of the unbuffered luminal medium was maintained at ∼7.4 by addition of strong acid or base. ATP stimulated base secretion, and this was inhibited by acetazolamide. Furthermore, these changes in secretion of base were in good quantitative agreement with the Isc responses. When phenol red (an estrogen) was removed from the culture medium, ATP-dependent HCO 3 - secretion was markedly reduced but could be restored by treatment with estradiol. Estrogens also markedly increased ciliation of the cultures. These results suggest that the midcycle increase in the HCO 3 - concentration of oviductal liquid may be mediated by the effects of estradiol on purinergic pathways or on ATP secretion.

AB - Because an increase in the HCO3 - concentration of oviductal liquid at midcycle is believed to markedly enhance fertility, we have studied active secretion of HCO3 - across highly differentiated cultures of monkey oviductal epithelium. Cultured cell sheets were mounted in Ussing chambers and bathed in medium containing 25 mM HCO 3 -. Purinergic agents potently stimulated short-circuit current (Isc) with an initial transient response declining within ∼2 min to a sustained response. The potency sequence of ATP ≈ UTP > ADP >> AMP suggested that the Isc response was mediated mainly by P2Y2 receptors. Acetazolamide, an inhibitor of carbonic anhydrase, had little or no effect on baseline Isc or the transient response to ATP but abolished the sustained response to ATP. Similar results were obtained on sheets of native epithelium. In pH-stat experiments, the abluminal medium of cell cultures was bathed in HCO3 --CO2 medium, and the pH of the unbuffered luminal medium was maintained at ∼7.4 by addition of strong acid or base. ATP stimulated base secretion, and this was inhibited by acetazolamide. Furthermore, these changes in secretion of base were in good quantitative agreement with the Isc responses. When phenol red (an estrogen) was removed from the culture medium, ATP-dependent HCO 3 - secretion was markedly reduced but could be restored by treatment with estradiol. Estrogens also markedly increased ciliation of the cultures. These results suggest that the midcycle increase in the HCO 3 - concentration of oviductal liquid may be mediated by the effects of estradiol on purinergic pathways or on ATP secretion.

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