CFTR mediates electrogenic chloride secretion in mouse inner medullary collecting duct (mIMCD-K2) cells

D. Vandorpe, N. Kizer, F. Ciampollilo, B. Moyer, K. Karlson, William B Guggino, B. A. Stanton

Research output: Contribution to journalArticle

Abstract

Previously we demonstrated that the inner medullary collecting duct cell line mIMCD-K2 secretes Cl- by an electrogenic mechanism [N. L. Kizer, B. Lewis, and B. A. Stanton. Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F347-F355, 1995; N. L. Kizer, D. Vandorpe, B. Lewis, B. Bunting, J. Russell, and B. A. Stanton. Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F854-F861, 1995]. The goal of the present study was to characterize the Cl- channel responsible for adenosine 3',5'-cyclic monophosphate (cAMP)stimulated Cl- secretion. To this end, using the patch- clamp technique, we measured Cl- currents. In whole cell patch-clamp experiments, 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate (CPT- cAMP) activated Cl- currents that were time and voltage independent, inhibited by diphenylamine 2-carboxylate (DPC), and had a linear current- voltage (I-V) relation. In cell-attached patches of the apical membrane, we identified 7-pS Cl- channels that were stimulated by CPT-cAMP. In inside- out patches with Cl- in the pipette and bath solutions, Cl- currents had a linear I-V relation. The halide permeability sequence was P(Cl) = P(Br) > P(I). The Cl--channel inhibitors DPC, 5-nitro-2-(3-phenylpropylamino)- benzoic acid, and glibenclamide blocked the 7-pS Cl- channel, whereas 4,4'- diisothiocyanostilbene-2,2'-disulfonic acid was ineffective. By reverse transcriptase polymerase chain reaction, we isolated a partial cDNA clone encoding the cystic fibrosis transmembrane conductance regulator in mIMCD-K2 cells. We conclude that cAMP stimulates electrogenic Cl- secretion in inner medullary collecting duct cells by activating cystic fibrosis transmembrane conductance regulator Cl- channels.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume269
Issue number3 38-3
StatePublished - 1995
Externally publishedYes

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Clamping devices
Cyclic AMP
Ducts
Electrolytes
Chlorides
chlorides
secretion
diphenylamine
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
Fluids
Glyburide
cystic fibrosis
Polymerase chain reaction
RNA-Directed DNA Polymerase
mice
cyclic AMP
Electric potential
electrolytes
Complementary DNA

Keywords

  • adenosine 3',5'-cyclic monophosphate
  • chloride conductance
  • cystic fibrosis transmembrane conductance regulator
  • kidney
  • nephron
  • patch clamp

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Agricultural and Biological Sciences(all)

Cite this

CFTR mediates electrogenic chloride secretion in mouse inner medullary collecting duct (mIMCD-K2) cells. / Vandorpe, D.; Kizer, N.; Ciampollilo, F.; Moyer, B.; Karlson, K.; Guggino, William B; Stanton, B. A.

In: American Journal of Physiology - Cell Physiology, Vol. 269, No. 3 38-3, 1995.

Research output: Contribution to journalArticle

Vandorpe, D. ; Kizer, N. ; Ciampollilo, F. ; Moyer, B. ; Karlson, K. ; Guggino, William B ; Stanton, B. A. / CFTR mediates electrogenic chloride secretion in mouse inner medullary collecting duct (mIMCD-K2) cells. In: American Journal of Physiology - Cell Physiology. 1995 ; Vol. 269, No. 3 38-3.
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AU - Vandorpe, D.

AU - Kizer, N.

AU - Ciampollilo, F.

AU - Moyer, B.

AU - Karlson, K.

AU - Guggino, William B

AU - Stanton, B. A.

PY - 1995

Y1 - 1995

N2 - Previously we demonstrated that the inner medullary collecting duct cell line mIMCD-K2 secretes Cl- by an electrogenic mechanism [N. L. Kizer, B. Lewis, and B. A. Stanton. Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F347-F355, 1995; N. L. Kizer, D. Vandorpe, B. Lewis, B. Bunting, J. Russell, and B. A. Stanton. Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F854-F861, 1995]. The goal of the present study was to characterize the Cl- channel responsible for adenosine 3',5'-cyclic monophosphate (cAMP)stimulated Cl- secretion. To this end, using the patch- clamp technique, we measured Cl- currents. In whole cell patch-clamp experiments, 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate (CPT- cAMP) activated Cl- currents that were time and voltage independent, inhibited by diphenylamine 2-carboxylate (DPC), and had a linear current- voltage (I-V) relation. In cell-attached patches of the apical membrane, we identified 7-pS Cl- channels that were stimulated by CPT-cAMP. In inside- out patches with Cl- in the pipette and bath solutions, Cl- currents had a linear I-V relation. The halide permeability sequence was P(Cl) = P(Br) > P(I). The Cl--channel inhibitors DPC, 5-nitro-2-(3-phenylpropylamino)- benzoic acid, and glibenclamide blocked the 7-pS Cl- channel, whereas 4,4'- diisothiocyanostilbene-2,2'-disulfonic acid was ineffective. By reverse transcriptase polymerase chain reaction, we isolated a partial cDNA clone encoding the cystic fibrosis transmembrane conductance regulator in mIMCD-K2 cells. We conclude that cAMP stimulates electrogenic Cl- secretion in inner medullary collecting duct cells by activating cystic fibrosis transmembrane conductance regulator Cl- channels.

AB - Previously we demonstrated that the inner medullary collecting duct cell line mIMCD-K2 secretes Cl- by an electrogenic mechanism [N. L. Kizer, B. Lewis, and B. A. Stanton. Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F347-F355, 1995; N. L. Kizer, D. Vandorpe, B. Lewis, B. Bunting, J. Russell, and B. A. Stanton. Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F854-F861, 1995]. The goal of the present study was to characterize the Cl- channel responsible for adenosine 3',5'-cyclic monophosphate (cAMP)stimulated Cl- secretion. To this end, using the patch- clamp technique, we measured Cl- currents. In whole cell patch-clamp experiments, 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate (CPT- cAMP) activated Cl- currents that were time and voltage independent, inhibited by diphenylamine 2-carboxylate (DPC), and had a linear current- voltage (I-V) relation. In cell-attached patches of the apical membrane, we identified 7-pS Cl- channels that were stimulated by CPT-cAMP. In inside- out patches with Cl- in the pipette and bath solutions, Cl- currents had a linear I-V relation. The halide permeability sequence was P(Cl) = P(Br) > P(I). The Cl--channel inhibitors DPC, 5-nitro-2-(3-phenylpropylamino)- benzoic acid, and glibenclamide blocked the 7-pS Cl- channel, whereas 4,4'- diisothiocyanostilbene-2,2'-disulfonic acid was ineffective. By reverse transcriptase polymerase chain reaction, we isolated a partial cDNA clone encoding the cystic fibrosis transmembrane conductance regulator in mIMCD-K2 cells. We conclude that cAMP stimulates electrogenic Cl- secretion in inner medullary collecting duct cells by activating cystic fibrosis transmembrane conductance regulator Cl- channels.

KW - adenosine 3',5'-cyclic monophosphate

KW - chloride conductance

KW - cystic fibrosis transmembrane conductance regulator

KW - kidney

KW - nephron

KW - patch clamp

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