Abstract
| Language | English |
|---|---|
| Pages | 633-644 |
| Number of pages | 11 |
| Journal | British Journal of Pharmacology |
| Volume | 157 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Jun 2009 |
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Keywords
- cAMP
- dominant negative
- glucagon-like peptide-1
- GLUTag cells
- H89
- PDE4D4
- phosphodiesterase
- rolipram
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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 journal › Article
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.
PY - 2009/6
Y1 - 2009/6
N2 - 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.
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
UR - http://dx.doi.org/10.1111/j.1476-5381.2009.00194.x
U2 - 10.1111/j.1476-5381.2009.00194.x
DO - 10.1111/j.1476-5381.2009.00194.x
M3 - Article
VL - 157
SP - 633
EP - 644
JO - British Journal of Pharmacology
T2 - British Journal of Pharmacology
JF - British Journal of Pharmacology
SN - 0007-1188
IS - 4
ER -