CGMP stimulates sodium and chloride currents in rat tracheal airway epithelia

Erik M. Schwiebert, Elizabeth D. Potter, Tae Ho Hwang, Jae Suk Woo, Changlin Ding, Weiping Qiu, William B Guggino, Michael A. Levine, Sandra E. Guggino

Research output: Contribution to journalArticle

Abstract

To test the hypothesis that guanosine 3',5'-cyclic monophosphate (cGMP) regulates ion transport in airway epithelial cells, we measured short- circuit current (I(SC)) and 22Na+ fluxes in primary cultured rat tracheal epithelial cells. In Cl--containing Ringer solution, I(SC) was increased by ~17 μA/cm2 after application of 1 mM 8-bromoguanosine 3',5'-cyclic monophosphate (8-BrcGMP), whereas, in Cl--free solutions, the Na+-mediated component was ~5 μA/cm2, suggesting a cGMP stimulation of Cl- secretory current and a smaller Na+ absorptive current. Inward and net mucosal-to- serosal 22Na+ flux was doubled in the presence of 2 mM 8-BrcGMP. To determine whether nucleotide-gated channels play a role in this transepithelial Na+ absorption, blockers of nucleotide-gated cation channels were used to inhibit I(SC). The cGMP-stimulated Na+ -mediated I(SC) was blocked by as little as 500 nM dichlorobenzamil or 50 μM L-cis-diltiazem, which are known blockers for cyclic nucleotide-gated cation channels. These agents also blocked the basal (non-cGMP-stimulated) current when measured in the presence of 10 μM amiloride, which blocks current through 5-pS amiloride-sensitive Na+ channels. To document whether the distribution of nucleotide-gated nonselective cation channels was consistent with a role in airway epithelial transport, in situ hybridization was performed. In situ hybridization of mRNA encoding for nucleotide-gated cation channels was found in epithelial cell layers of rat trachea, bronchi, bronchioles, and alveolar cells but not in smooth muscle layers or tracheal cartilage. Reverse transcriptase-polymerase chain reaction, restriction enzyme analysis, and sequencing of the cDNA transcribed from mRNA of whole lung and tracheal epithelial cells indicate that a channel highly homologous to the retinal nucleotide-gated nonselective cation channel (CNG1) is present. Thus these data, along with evidence supporting the existence of signal transduction pathways elevating intracellular levels of cGMP, indicate that cGMP regulates transepithelial ion transport in lung epithelial tissues.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume272
Issue number3 41-3
StatePublished - Mar 1997

Fingerprint

Sodium Chloride
Rats
Epithelium
Nucleotides
Cations
Epithelial Cells
Amiloride
Ion Transport
In Situ Hybridization
Cyclic Nucleotide-Gated Cation Channels
Ions
Fluxes
Alveolar Epithelial Cells
Bronchioles
Lung
Signal transduction
Restriction Mapping
Messenger RNA
Diltiazem
Polymerase chain reaction

Keywords

  • cyclic nucleotide- gated cation channels
  • cystic fibrosis
  • guanosine 3',5'-cyclic monophosphate

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Physiology (medical)

Cite this

Schwiebert, E. M., Potter, E. D., Hwang, T. H., Woo, J. S., Ding, C., Qiu, W., ... Guggino, S. E. (1997). CGMP stimulates sodium and chloride currents in rat tracheal airway epithelia. American Journal of Physiology - Cell Physiology, 272(3 41-3).

CGMP stimulates sodium and chloride currents in rat tracheal airway epithelia. / Schwiebert, Erik M.; Potter, Elizabeth D.; Hwang, Tae Ho; Woo, Jae Suk; Ding, Changlin; Qiu, Weiping; Guggino, William B; Levine, Michael A.; Guggino, Sandra E.

In: American Journal of Physiology - Cell Physiology, Vol. 272, No. 3 41-3, 03.1997.

Research output: Contribution to journalArticle

Schwiebert, EM, Potter, ED, Hwang, TH, Woo, JS, Ding, C, Qiu, W, Guggino, WB, Levine, MA & Guggino, SE 1997, 'CGMP stimulates sodium and chloride currents in rat tracheal airway epithelia', American Journal of Physiology - Cell Physiology, vol. 272, no. 3 41-3.
Schwiebert EM, Potter ED, Hwang TH, Woo JS, Ding C, Qiu W et al. CGMP stimulates sodium and chloride currents in rat tracheal airway epithelia. American Journal of Physiology - Cell Physiology. 1997 Mar;272(3 41-3).
Schwiebert, Erik M. ; Potter, Elizabeth D. ; Hwang, Tae Ho ; Woo, Jae Suk ; Ding, Changlin ; Qiu, Weiping ; Guggino, William B ; Levine, Michael A. ; Guggino, Sandra E. / CGMP stimulates sodium and chloride currents in rat tracheal airway epithelia. In: American Journal of Physiology - Cell Physiology. 1997 ; Vol. 272, No. 3 41-3.
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AU - Ding, Changlin

AU - Qiu, Weiping

AU - Guggino, William B

AU - Levine, Michael A.

AU - Guggino, Sandra E.

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N2 - To test the hypothesis that guanosine 3',5'-cyclic monophosphate (cGMP) regulates ion transport in airway epithelial cells, we measured short- circuit current (I(SC)) and 22Na+ fluxes in primary cultured rat tracheal epithelial cells. In Cl--containing Ringer solution, I(SC) was increased by ~17 μA/cm2 after application of 1 mM 8-bromoguanosine 3',5'-cyclic monophosphate (8-BrcGMP), whereas, in Cl--free solutions, the Na+-mediated component was ~5 μA/cm2, suggesting a cGMP stimulation of Cl- secretory current and a smaller Na+ absorptive current. Inward and net mucosal-to- serosal 22Na+ flux was doubled in the presence of 2 mM 8-BrcGMP. To determine whether nucleotide-gated channels play a role in this transepithelial Na+ absorption, blockers of nucleotide-gated cation channels were used to inhibit I(SC). The cGMP-stimulated Na+ -mediated I(SC) was blocked by as little as 500 nM dichlorobenzamil or 50 μM L-cis-diltiazem, which are known blockers for cyclic nucleotide-gated cation channels. These agents also blocked the basal (non-cGMP-stimulated) current when measured in the presence of 10 μM amiloride, which blocks current through 5-pS amiloride-sensitive Na+ channels. To document whether the distribution of nucleotide-gated nonselective cation channels was consistent with a role in airway epithelial transport, in situ hybridization was performed. In situ hybridization of mRNA encoding for nucleotide-gated cation channels was found in epithelial cell layers of rat trachea, bronchi, bronchioles, and alveolar cells but not in smooth muscle layers or tracheal cartilage. Reverse transcriptase-polymerase chain reaction, restriction enzyme analysis, and sequencing of the cDNA transcribed from mRNA of whole lung and tracheal epithelial cells indicate that a channel highly homologous to the retinal nucleotide-gated nonselective cation channel (CNG1) is present. Thus these data, along with evidence supporting the existence of signal transduction pathways elevating intracellular levels of cGMP, indicate that cGMP regulates transepithelial ion transport in lung epithelial tissues.

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