The role of the PDE4D cAMP phosphodiesterase in the regulation of glucagon-like peptide-1 release

W.K. Ong, F.M. Gribble, F. Reimann, M.J. Lynch, M.D. Houslay, G.S. Baillie, B.L. Furman, N.J. Pyne

Research output: Contribution to journalArticle

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Abstract

Increases in intracellular cyclic AMP (cAMP) augment the release/secretion of glucagon-like peptide-1 (GLP-1). As cAMP is hydrolysed by cAMP phosphodiesterases (PDEs), we determined the role of PDEs and particularly PDE4 in regulating GLP-1 release. GLP-1 release, PDE expression and activity were investigated using rats and GLUTag cells, a GLP-1-releasing cell line. The effects of rolipram, a selective PDE4 inhibitor both in vivo and in vitro and stably overexpressed catalytically inactive PDE4D5 (D556A-PDE4D5) mutant in vitro on GLP-1 release were investigated. Rolipram (1.5 mg.kg(-1) i.v.) increased plasma GLP-1 concentrations approximately twofold above controls in anaesthetized rats and enhanced glucose-induced GLP-1 release in GLUTag cells (EC50 similar to 1.2 nmol.L-1). PDE4D mRNA transcript and protein were detected in GLUTag cells using RT-PCR with gene-specific primers and Western blotting with a specific PDE4D antibody respectively. Moreover, significant PDE activity was inhibited by rolipram in GLUTag cells. A GLUTag cell clone (C1) stably overexpressing the D556A-PDE4D5 mutant, exhibited elevated intracellular cAMP levels and increased basal and glucose-induced GLP-1 release compared with vector-transfected control cells. A role for intracellular cAMP/PKA in enhancing GLP-1 release in response to overexpression of D556A-PDE4D5 mutant was demonstrated by the finding that the PKA inhibitor H89 reduced both basal and glucose-induced GLP-1 release by 37% and 39%, respectively, from C1 GLUTag cells. PDE4D may play an important role in regulating intracellular cAMP linked to the regulation of GLP-1 release.
LanguageEnglish
Pages633-644
Number of pages11
JournalBritish Journal of Pharmacology
Volume157
Issue number4
DOIs
Publication statusPublished - Jun 2009

Fingerprint

Glucagon-Like Peptide 1
Phosphoric Diester Hydrolases
Cyclic AMP
Rolipram
Glucose
Type 4 Cyclic Nucleotide Phosphodiesterase
Phosphodiesterase 4 Inhibitors
Clone Cells
Western Blotting

Keywords

  • cAMP
  • dominant negative
  • glucagon-like peptide-1
  • GLUTag cells
  • H89
  • PDE4D4
  • phosphodiesterase
  • rolipram

Cite this

Ong, W.K. ; Gribble, F.M. ; Reimann, F. ; Lynch, M.J. ; Houslay, M.D. ; Baillie, G.S. ; Furman, B.L. ; Pyne, N.J. / The role of the PDE4D cAMP phosphodiesterase in the regulation of glucagon-like peptide-1 release. In: British Journal of Pharmacology. 2009 ; Vol. 157, No. 4. pp. 633-644.
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The role of the PDE4D cAMP phosphodiesterase in the regulation of glucagon-like peptide-1 release. / Ong, W.K.; Gribble, F.M.; Reimann, F.; Lynch, M.J.; Houslay, M.D.; Baillie, G.S.; Furman, B.L.; Pyne, N.J.

In: British Journal of Pharmacology, Vol. 157, No. 4, 06.2009, p. 633-644.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The role of the PDE4D cAMP phosphodiesterase in the regulation of glucagon-like peptide-1 release

AU - Ong, W.K.

AU - Gribble, F.M.

AU - Reimann, F.

AU - Lynch, M.J.

AU - Houslay, M.D.

AU - Baillie, G.S.

AU - Furman, B.L.

AU - Pyne, N.J.

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AB - Increases in intracellular cyclic AMP (cAMP) augment the release/secretion of glucagon-like peptide-1 (GLP-1). As cAMP is hydrolysed by cAMP phosphodiesterases (PDEs), we determined the role of PDEs and particularly PDE4 in regulating GLP-1 release. GLP-1 release, PDE expression and activity were investigated using rats and GLUTag cells, a GLP-1-releasing cell line. The effects of rolipram, a selective PDE4 inhibitor both in vivo and in vitro and stably overexpressed catalytically inactive PDE4D5 (D556A-PDE4D5) mutant in vitro on GLP-1 release were investigated. Rolipram (1.5 mg.kg(-1) i.v.) increased plasma GLP-1 concentrations approximately twofold above controls in anaesthetized rats and enhanced glucose-induced GLP-1 release in GLUTag cells (EC50 similar to 1.2 nmol.L-1). PDE4D mRNA transcript and protein were detected in GLUTag cells using RT-PCR with gene-specific primers and Western blotting with a specific PDE4D antibody respectively. Moreover, significant PDE activity was inhibited by rolipram in GLUTag cells. A GLUTag cell clone (C1) stably overexpressing the D556A-PDE4D5 mutant, exhibited elevated intracellular cAMP levels and increased basal and glucose-induced GLP-1 release compared with vector-transfected control cells. A role for intracellular cAMP/PKA in enhancing GLP-1 release in response to overexpression of D556A-PDE4D5 mutant was demonstrated by the finding that the PKA inhibitor H89 reduced both basal and glucose-induced GLP-1 release by 37% and 39%, respectively, from C1 GLUTag cells. PDE4D may play an important role in regulating intracellular cAMP linked to the regulation of GLP-1 release.

KW - cAMP

KW - dominant negative

KW - glucagon-like peptide-1

KW - GLUTag cells

KW - H89

KW - PDE4D4

KW - phosphodiesterase

KW - rolipram

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