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Article: Electrophysiological studies of anion secretion in cultured human epididymal cells

TitleElectrophysiological studies of anion secretion in cultured human epididymal cells
Authors
Issue Date1992
PublisherWiley-Blackwell Publishing Ltd.. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=0022-3751
Citation
Journal Of Physiology, 1992, v. 455, p. 455-469 How to Cite?
Abstract1. Primary monolayer cultures from adult human epididymis were grown on Petri dishes and pervious supports. The epithelia so formed were used for whole-cell patch clamp recording and short-circuit current (I(SC)) measurement. 2. After 50 days of culture, the cells formed a tight epithelium with transepithelial potential of 5.6 ± 1.3 mV (mean ± S.E.M., n = 16), apical side negative, and a basal I(SC) Of 6.9 ± 0.9 μA cm -2 (mean ± S.E.M., n = 16). 3. Adrenaline, when added to the basolateral side, at a concentration of 0.23 μmol 1 -1 increased the I(SC) by 3.0 ± 1.2 μA cm -2 (mean ± S.E.M., n = 4). This increase was blockable by diphenylamine-2-carboxylate (DPC, 1 mmol 1 -1. Forskolin (10 μmol l -1) also evoked a similar response to adrenaline. 4. In whole-cell patch clamp experiment, the resting membrane potential of the cells after dialysis with pipette solution containing 135 mmol l -1 KCl was found to be -30 ± 14 mV (mean ± S.E.M., n = 15). 5. About 90% of the cells successfully forming patches responded to 1 μmol l -1 adrenaline by an increase in inward current at -70 mV holding potential (ΔI = -1600 ± 900 pA, mean ± S.E.M., n = 15). This increase in current was accompanied by a shift in reversal potential to -2 ± 1 mV (mean ± S.E.M., n = 16). 6. The adrenaline-induced inward current was found to be blockable by the Cl - channel blocker, DPC (0.25 mmol l -1). Ion substitution experiments showed that the adrenaline-evoked current was carried mainly by Cl -. 7. The effect of adrenaline on thec whole-cell current was found to be mimicked by forskolin and could be abolished by including GDPβS or a protein kinase A inhibitor in the pipette solution. Propranolol, but not phentolamine, completely abolished the effect of adrenaline. 8. Inclusion of 20 mmol l -1 EGTA or 2 mmol l -1 BAPTA+ 100 μmol l -1 TMB-8 (to inhibit intracellular Ca 2+ release) in the pipette did not seem to have any marked effect on adrenaline-evoked whole-cell current. Lowering the pipette Ca 2+ concentration to 1 nmol l -1 or raising it to 10 μmol l -1 had no effect on the whole-cell current response to adrenaline. 9. This study shows that adrenaline stimulates Cl - secretion in cultured human epididymal cells. The major intracellular mechanism appears to involve the classical β-adrenoceptor pathway, viz. β-adrenoceptor G(s) protein-adenylate cyclase cyclic AMP protein kinase A. However, the role of Ca 2+ in secretion in the epididymis and its interaction with cyclic AMP awaits clarification.
Persistent Identifierhttp://hdl.handle.net/10722/161939
ISSN
2015 Impact Factor: 4.731
2015 SCImago Journal Rankings: 2.670
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorHuang, SJen_US
dc.contributor.authorLeung, AYHen_US
dc.contributor.authorFu, WOen_US
dc.contributor.authorChung, YWen_US
dc.contributor.authorZhou, TSen_US
dc.contributor.authorChan, PSFen_US
dc.contributor.authorWong, PYDen_US
dc.date.accessioned2012-09-05T05:16:10Z-
dc.date.available2012-09-05T05:16:10Z-
dc.date.issued1992en_US
dc.identifier.citationJournal Of Physiology, 1992, v. 455, p. 455-469en_US
dc.identifier.issn0022-3751en_US
dc.identifier.urihttp://hdl.handle.net/10722/161939-
dc.description.abstract1. Primary monolayer cultures from adult human epididymis were grown on Petri dishes and pervious supports. The epithelia so formed were used for whole-cell patch clamp recording and short-circuit current (I(SC)) measurement. 2. After 50 days of culture, the cells formed a tight epithelium with transepithelial potential of 5.6 ± 1.3 mV (mean ± S.E.M., n = 16), apical side negative, and a basal I(SC) Of 6.9 ± 0.9 μA cm -2 (mean ± S.E.M., n = 16). 3. Adrenaline, when added to the basolateral side, at a concentration of 0.23 μmol 1 -1 increased the I(SC) by 3.0 ± 1.2 μA cm -2 (mean ± S.E.M., n = 4). This increase was blockable by diphenylamine-2-carboxylate (DPC, 1 mmol 1 -1. Forskolin (10 μmol l -1) also evoked a similar response to adrenaline. 4. In whole-cell patch clamp experiment, the resting membrane potential of the cells after dialysis with pipette solution containing 135 mmol l -1 KCl was found to be -30 ± 14 mV (mean ± S.E.M., n = 15). 5. About 90% of the cells successfully forming patches responded to 1 μmol l -1 adrenaline by an increase in inward current at -70 mV holding potential (ΔI = -1600 ± 900 pA, mean ± S.E.M., n = 15). This increase in current was accompanied by a shift in reversal potential to -2 ± 1 mV (mean ± S.E.M., n = 16). 6. The adrenaline-induced inward current was found to be blockable by the Cl - channel blocker, DPC (0.25 mmol l -1). Ion substitution experiments showed that the adrenaline-evoked current was carried mainly by Cl -. 7. The effect of adrenaline on thec whole-cell current was found to be mimicked by forskolin and could be abolished by including GDPβS or a protein kinase A inhibitor in the pipette solution. Propranolol, but not phentolamine, completely abolished the effect of adrenaline. 8. Inclusion of 20 mmol l -1 EGTA or 2 mmol l -1 BAPTA+ 100 μmol l -1 TMB-8 (to inhibit intracellular Ca 2+ release) in the pipette did not seem to have any marked effect on adrenaline-evoked whole-cell current. Lowering the pipette Ca 2+ concentration to 1 nmol l -1 or raising it to 10 μmol l -1 had no effect on the whole-cell current response to adrenaline. 9. This study shows that adrenaline stimulates Cl - secretion in cultured human epididymal cells. The major intracellular mechanism appears to involve the classical β-adrenoceptor pathway, viz. β-adrenoceptor G(s) protein-adenylate cyclase cyclic AMP protein kinase A. However, the role of Ca 2+ in secretion in the epididymis and its interaction with cyclic AMP awaits clarification.en_US
dc.languageengen_US
dc.publisherWiley-Blackwell Publishing Ltd.. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=0022-3751en_US
dc.relation.ispartofJournal of Physiologyen_US
dc.subject.meshAdrenergic Agonists - Pharmacologyen_US
dc.subject.meshCalcium - Metabolismen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshChlorides - Metabolismen_US
dc.subject.meshCyclic Amp - Metabolismen_US
dc.subject.meshEpididymis - Secretionen_US
dc.subject.meshEpinephrine - Metabolismen_US
dc.subject.meshHumansen_US
dc.subject.meshMaleen_US
dc.subject.meshMembrane Potentials - Physiologyen_US
dc.titleElectrophysiological studies of anion secretion in cultured human epididymal cellsen_US
dc.typeArticleen_US
dc.identifier.emailLeung, AYH:ayhleung@hku.hken_US
dc.identifier.authorityLeung, AYH=rp00265en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid1362444en_US
dc.identifier.scopuseid_2-s2.0-0026660655en_US
dc.identifier.volume455en_US
dc.identifier.spage455en_US
dc.identifier.epage469en_US
dc.identifier.isiWOS:A1992JN28700025-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridHuang, SJ=7405416278en_US
dc.identifier.scopusauthoridLeung, AYH=7403012668en_US
dc.identifier.scopusauthoridFu, WO=7202947290en_US
dc.identifier.scopusauthoridChung, YW=7404388001en_US
dc.identifier.scopusauthoridZhou, TS=7402989554en_US
dc.identifier.scopusauthoridChan, PSF=7403497783en_US
dc.identifier.scopusauthoridWong, PYD=7403980262en_US

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