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Functional expression of cystic fibrosis transmembrane conductance regulator in rat oviduct epithelium.

Abstract

The aim of this study was to investigate the functional expression of cystic fibrosis transmembrane conductance regulator (CFTR) with electrophysiological and molecular technique in rat oviduct epithelium. In whole-cell patch clamp, oviduct epithelial cells responded to 100 microM 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP) with a rise in inward current in Gap-free mode, which was inhibited successively by 5 microM CFTR(inh)-172, a CFTR specific inhibitor, and 1 mM diphenylamine-2-carboxylate (DPC), the Cl- channel blocker. The cAMP-activated current exhibited a linear current-voltage (I-V) relationship and time- and voltage-independent characteristics. The reversal potentials of the cAMP-activated currents in symmetrical Cl- solutions were close to the Cl- equilibrium, 0.5+/-0.2 mV (n=4). When Cl- concentration in the bath solution was changed from 140 mM to 70 mM and a pipette solution containing 140 mM Cl- was used, the reversal potential shifted to a value close to the new equilibrium for Cl-, 20+/-0.6 mV (n=4), as compared with the theoretic value of 18.7 mV. In addition, mRNA expression of CFTR was also detected in rat oviduct epithelium. Western blot analysis showed that CFTR protein is found in the oviduct throughout the cycle with maximal expression at estrus, and immunofluorescence and immunohistochemistry analysis revealed that CFTR is located at the apical membrane of the epithelial cells. These results showed that the cAMP-activated Cl- current in the oviduct epithelium was characteristic of CFTR, which provided direct evidence for the functional expression of CFTR in the rat oviduct epithelium. CFTR may play a role in modulating fluid transport in the oviduct.

Authors+Show Affiliations

School of Life Science, Sun Yat-sen University, Guangzhou 510275, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18850051

Citation

Chen, Minhui, et al. "Functional Expression of Cystic Fibrosis Transmembrane Conductance Regulator in Rat Oviduct Epithelium." Acta Biochimica Et Biophysica Sinica, vol. 40, no. 10, 2008, pp. 864-72.
Chen M, Du J, Jiang W, et al. Functional expression of cystic fibrosis transmembrane conductance regulator in rat oviduct epithelium. Acta Biochim Biophys Sin (Shanghai). 2008;40(10):864-72.
Chen, M., Du, J., Jiang, W., Zuo, W., Wang, F., Li, M., ... Zhou, W. (2008). Functional expression of cystic fibrosis transmembrane conductance regulator in rat oviduct epithelium. Acta Biochimica Et Biophysica Sinica, 40(10), pp. 864-72.
Chen M, et al. Functional Expression of Cystic Fibrosis Transmembrane Conductance Regulator in Rat Oviduct Epithelium. Acta Biochim Biophys Sin (Shanghai). 2008;40(10):864-72. PubMed PMID: 18850051.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Functional expression of cystic fibrosis transmembrane conductance regulator in rat oviduct epithelium. AU - Chen,Minhui, AU - Du,Jianyang, AU - Jiang,Weijian, AU - Zuo,Wulin, AU - Wang,Fang, AU - Li,Manhui, AU - Chan,Hsiaochang, AU - Zhou,Wenliang, PY - 2008/10/14/pubmed PY - 2008/12/17/medline PY - 2008/10/14/entrez SP - 864 EP - 72 JF - Acta biochimica et biophysica Sinica JO - Acta Biochim. Biophys. Sin. (Shanghai) VL - 40 IS - 10 N2 - The aim of this study was to investigate the functional expression of cystic fibrosis transmembrane conductance regulator (CFTR) with electrophysiological and molecular technique in rat oviduct epithelium. In whole-cell patch clamp, oviduct epithelial cells responded to 100 microM 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP) with a rise in inward current in Gap-free mode, which was inhibited successively by 5 microM CFTR(inh)-172, a CFTR specific inhibitor, and 1 mM diphenylamine-2-carboxylate (DPC), the Cl- channel blocker. The cAMP-activated current exhibited a linear current-voltage (I-V) relationship and time- and voltage-independent characteristics. The reversal potentials of the cAMP-activated currents in symmetrical Cl- solutions were close to the Cl- equilibrium, 0.5+/-0.2 mV (n=4). When Cl- concentration in the bath solution was changed from 140 mM to 70 mM and a pipette solution containing 140 mM Cl- was used, the reversal potential shifted to a value close to the new equilibrium for Cl-, 20+/-0.6 mV (n=4), as compared with the theoretic value of 18.7 mV. In addition, mRNA expression of CFTR was also detected in rat oviduct epithelium. Western blot analysis showed that CFTR protein is found in the oviduct throughout the cycle with maximal expression at estrus, and immunofluorescence and immunohistochemistry analysis revealed that CFTR is located at the apical membrane of the epithelial cells. These results showed that the cAMP-activated Cl- current in the oviduct epithelium was characteristic of CFTR, which provided direct evidence for the functional expression of CFTR in the rat oviduct epithelium. CFTR may play a role in modulating fluid transport in the oviduct. SN - 1745-7270 UR - https://www.unboundmedicine.com/medline/citation/18850051/Functional_expression_of_cystic_fibrosis_transmembrane_conductance_regulator_in_rat_oviduct_epithelium_ L2 - http://www.abbs.org.cn/fulltxt/40-10/40100864.htm DB - PRIME DP - Unbound Medicine ER -