Protein kinase A and mitogen-activated protein kinase pathways mediate cAMP induction of α-epithelial Na+ channels (α-ENaC)

Shamimunisa B Mustafa, Robert Castro, Alison J. Falck, Jean O Petershack, Barbara M. Henson, Yvonne M. Mendoza, Ahsan Choudary, Steven R Seidner

Research output: Contribution to journalArticle

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Abstract

A major mechanism for Na+ transport across epithelia occurs through epithelial Na+ channels (ENaC). ENaC is a multimeric channel consisting of three subunits (α, β, and γ). The α-subunit is critical for ENaC function. In specific culture conditions, the rat submandibular gland epithelial cell line (SMG-C6) demonstrates minimal Na + transport properties and exposure to dibutyryl cAMP (DbcAMP) for up to 48 h caused an elevation of α-ENaC mRNA and protein expression and amiloride-sensitive short-circuit current (ISC). Here we examined the early signaling pathways evoked by DbcAMP which contribute to the eventual increase in Na+ transport. Treatment with either of the protein kinase A (PKA) inhibitors KT5720 or H-89 followed by exposure to 1 mM DbcAMP for 24 h markedly attenuated DbcAMP-induced α-ENaC protein formation and ISC. Exposure of SMG-C6 cells to 1 mM DbcAMP induced a rapid, transient phosphorylation of the cAMP response element binding protein (CREB). This response was attenuated in the presence of either KT5720 or H-89. Dominant-negative CREB decreased DbcAMP-induced α-ENaC expression. Suppression of the extracellular signal-regulated protein kinase (ERK 1,2) with PD98059 or the p38 mitogen-activated protein kinase (MAPK) pathway with SB203580 reduced DbcAMP-induced α-ENaC protein levels in SMG-C6 cells. DbcAMP-induced phosphorylation of CREB was markedly attenuated by PD98059 or SB203580. DbcAMP-induced activation of the either the p38 or the ERK 1,2 MAPK pathways was abolished by either of the PKA inhibitors, H-89 or KT5720. Cross talk between these signaling pathways induced by DbcAMP via the activation of CREB appears to contribute to increased levels of α-ENaC observed after 24 h of treatment in SMG-C6 epithelial cells.

Original languageEnglish (US)
Pages (from-to)101-110
Number of pages10
JournalJournal of Cellular Physiology
Volume215
Issue number1
DOIs
StatePublished - Apr 2008

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Epithelial Sodium Channels
Cyclic AMP-Dependent Protein Kinases
Mitogen-Activated Protein Kinases
Cyclic AMP Response Element-Binding Protein
Phosphorylation
Protein Kinase Inhibitors
Epithelial Cells
Chemical activation
Proteins
Amiloride
Submandibular Gland
Mitogen-Activated Protein Kinase 1
Extracellular Signal-Regulated MAP Kinases
p38 Mitogen-Activated Protein Kinases
Short circuit currents
Transport properties
Protein Kinases
Rats
Epithelium
Cell Line

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Protein kinase A and mitogen-activated protein kinase pathways mediate cAMP induction of α-epithelial Na+ channels (α-ENaC). / Mustafa, Shamimunisa B; Castro, Robert; Falck, Alison J.; Petershack, Jean O; Henson, Barbara M.; Mendoza, Yvonne M.; Choudary, Ahsan; Seidner, Steven R.

In: Journal of Cellular Physiology, Vol. 215, No. 1, 04.2008, p. 101-110.

Research output: Contribution to journalArticle

Mustafa, SB, Castro, R, Falck, AJ, Petershack, JO, Henson, BM, Mendoza, YM, Choudary, A & Seidner, SR 2008, 'Protein kinase A and mitogen-activated protein kinase pathways mediate cAMP induction of α-epithelial Na+ channels (α-ENaC)', Journal of Cellular Physiology, vol. 215, no. 1, pp. 101-110. https://doi.org/10.1002/jcp.21291
Mustafa, Shamimunisa B ; Castro, Robert ; Falck, Alison J. ; Petershack, Jean O ; Henson, Barbara M. ; Mendoza, Yvonne M. ; Choudary, Ahsan ; Seidner, Steven R. / Protein kinase A and mitogen-activated protein kinase pathways mediate cAMP induction of α-epithelial Na+ channels (α-ENaC). In: Journal of Cellular Physiology. 2008 ; Vol. 215, No. 1. pp. 101-110.
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