beta-Adrenoceptor stimulation potentiates insulin-stimulated PKB phosphorylation in rat cardiomyocytes via cAMP and PKA
Authors
Stuenaes, Jorid T.Bolling, Astrid
Ingvaldsen, Ada
Rommundstad, Camilla
Sudar, Emina
Lin, Fang-Chin
Lai, Yu-Chiang
Jensen, Jorgen
Article
Metadata
Show full item recordAbstract
Background and purpose: Genetic approaches have documented protein kinase B (PKB) as a pivotal regulator of heart function. Insulin strongly activates PKB, whereas adrenaline is not considered a major physiological regulator of PKB in heart. In skeletal muscles, however, adrenaline potentiates insulin-stimulated PKB activation without having effect in the absence of insulin. The purpose of the present study was to investigate the interaction between insulin and beta-adrenergic stimulation in regulation of PKB phosphorylation. Experimental approach: Cardiomyocytes were isolated from adult rats by collagenase, and incubated with insulin, isoprenaline, and other compounds. Protein phosphorylation was evaluated by Western blot and phospho-specific antibodies. Key results: Isoprenaline increased insulin-stimulated PKB Ser473 and Thr308 phosphorylation more than threefold in cardiomyocytes. Isoprenaline alone did not increase PKB phosphorylation. Isoprenaline also increased insulin-stimulate...d GSK-3 beta Ser9 phosphorylation approximately twofold, supporting that PKB phosphorylation increased kinase activity. Dobutamine (beta(1)-agonist) increased insulin-stimulated PKB phosphorylation as effectively as isoprenaline (more than threefold), whereas salbutamol (beta(2)-agonist) only potentiated insulin-stimulated PKB phosphorylation by approximately 80%. Dobutamine, but not salbutamol, increased phospholamban Ser16 phosphorylation and glycogen phosphorylase activation (PKA-mediated effects). Furthermore, the cAMP analogue that activates PKA (dibutyryl-cAMP and N6-benzoyl-cAMP) increased insulin-stimulated PKB phosphorylation by more than threefold without effect alone. The Epac-specific activator 8-(4-chlorophenylthio)-2-O-methyl-cAMP (007) increased insulin-stimulated PKB phosphorylation by approximately 50%. Db-cAMP and N6-benzoyl-cAMP, but not 007, increased phospholamban Ser16 phosphorylation. Conclusions and implications: beta-adrenoceptors are strong regulators of PKB phosphorylation via cAMP and PKA when insulin is present. We hypothesize that PKB mediates important signalling in the heart during beta-adrenergic receptors stimulation.
Keywords:
Heart / Akt / GSK-3 / phosphorylation / phosphatidylinositol 3-kinase / phoshodiesterase / phospholamban / hypertrophy / rolipram / ERKSource:
British Journal of Pharmacology, 2010, 160, 1, 116-129Funding / projects:
- Novo Nordisk Foundation, European Commission [COST BM0602]
DOI: 10.1111/j.1476-5381.2010.00677.x
ISSN: 0007-1188
PubMed: 20412069
WoS: 000276651200011
Scopus: 2-s2.0-77950956173
Collections
Institution/Community
VinčaTY - JOUR AU - Stuenaes, Jorid T. AU - Bolling, Astrid AU - Ingvaldsen, Ada AU - Rommundstad, Camilla AU - Sudar, Emina AU - Lin, Fang-Chin AU - Lai, Yu-Chiang AU - Jensen, Jorgen PY - 2010 UR - https://vinar.vin.bg.ac.rs/handle/123456789/3977 AB - Background and purpose: Genetic approaches have documented protein kinase B (PKB) as a pivotal regulator of heart function. Insulin strongly activates PKB, whereas adrenaline is not considered a major physiological regulator of PKB in heart. In skeletal muscles, however, adrenaline potentiates insulin-stimulated PKB activation without having effect in the absence of insulin. The purpose of the present study was to investigate the interaction between insulin and beta-adrenergic stimulation in regulation of PKB phosphorylation. Experimental approach: Cardiomyocytes were isolated from adult rats by collagenase, and incubated with insulin, isoprenaline, and other compounds. Protein phosphorylation was evaluated by Western blot and phospho-specific antibodies. Key results: Isoprenaline increased insulin-stimulated PKB Ser473 and Thr308 phosphorylation more than threefold in cardiomyocytes. Isoprenaline alone did not increase PKB phosphorylation. Isoprenaline also increased insulin-stimulated GSK-3 beta Ser9 phosphorylation approximately twofold, supporting that PKB phosphorylation increased kinase activity. Dobutamine (beta(1)-agonist) increased insulin-stimulated PKB phosphorylation as effectively as isoprenaline (more than threefold), whereas salbutamol (beta(2)-agonist) only potentiated insulin-stimulated PKB phosphorylation by approximately 80%. Dobutamine, but not salbutamol, increased phospholamban Ser16 phosphorylation and glycogen phosphorylase activation (PKA-mediated effects). Furthermore, the cAMP analogue that activates PKA (dibutyryl-cAMP and N6-benzoyl-cAMP) increased insulin-stimulated PKB phosphorylation by more than threefold without effect alone. The Epac-specific activator 8-(4-chlorophenylthio)-2-O-methyl-cAMP (007) increased insulin-stimulated PKB phosphorylation by approximately 50%. Db-cAMP and N6-benzoyl-cAMP, but not 007, increased phospholamban Ser16 phosphorylation. Conclusions and implications: beta-adrenoceptors are strong regulators of PKB phosphorylation via cAMP and PKA when insulin is present. We hypothesize that PKB mediates important signalling in the heart during beta-adrenergic receptors stimulation. T2 - British Journal of Pharmacology T1 - beta-Adrenoceptor stimulation potentiates insulin-stimulated PKB phosphorylation in rat cardiomyocytes via cAMP and PKA VL - 160 IS - 1 SP - 116 EP - 129 DO - 10.1111/j.1476-5381.2010.00677.x ER -
@article{ author = "Stuenaes, Jorid T. and Bolling, Astrid and Ingvaldsen, Ada and Rommundstad, Camilla and Sudar, Emina and Lin, Fang-Chin and Lai, Yu-Chiang and Jensen, Jorgen", year = "2010", abstract = "Background and purpose: Genetic approaches have documented protein kinase B (PKB) as a pivotal regulator of heart function. Insulin strongly activates PKB, whereas adrenaline is not considered a major physiological regulator of PKB in heart. In skeletal muscles, however, adrenaline potentiates insulin-stimulated PKB activation without having effect in the absence of insulin. The purpose of the present study was to investigate the interaction between insulin and beta-adrenergic stimulation in regulation of PKB phosphorylation. Experimental approach: Cardiomyocytes were isolated from adult rats by collagenase, and incubated with insulin, isoprenaline, and other compounds. Protein phosphorylation was evaluated by Western blot and phospho-specific antibodies. Key results: Isoprenaline increased insulin-stimulated PKB Ser473 and Thr308 phosphorylation more than threefold in cardiomyocytes. Isoprenaline alone did not increase PKB phosphorylation. Isoprenaline also increased insulin-stimulated GSK-3 beta Ser9 phosphorylation approximately twofold, supporting that PKB phosphorylation increased kinase activity. Dobutamine (beta(1)-agonist) increased insulin-stimulated PKB phosphorylation as effectively as isoprenaline (more than threefold), whereas salbutamol (beta(2)-agonist) only potentiated insulin-stimulated PKB phosphorylation by approximately 80%. Dobutamine, but not salbutamol, increased phospholamban Ser16 phosphorylation and glycogen phosphorylase activation (PKA-mediated effects). Furthermore, the cAMP analogue that activates PKA (dibutyryl-cAMP and N6-benzoyl-cAMP) increased insulin-stimulated PKB phosphorylation by more than threefold without effect alone. The Epac-specific activator 8-(4-chlorophenylthio)-2-O-methyl-cAMP (007) increased insulin-stimulated PKB phosphorylation by approximately 50%. Db-cAMP and N6-benzoyl-cAMP, but not 007, increased phospholamban Ser16 phosphorylation. Conclusions and implications: beta-adrenoceptors are strong regulators of PKB phosphorylation via cAMP and PKA when insulin is present. We hypothesize that PKB mediates important signalling in the heart during beta-adrenergic receptors stimulation.", journal = "British Journal of Pharmacology", title = "beta-Adrenoceptor stimulation potentiates insulin-stimulated PKB phosphorylation in rat cardiomyocytes via cAMP and PKA", volume = "160", number = "1", pages = "116-129", doi = "10.1111/j.1476-5381.2010.00677.x" }
Stuenaes, J. T., Bolling, A., Ingvaldsen, A., Rommundstad, C., Sudar, E., Lin, F., Lai, Y.,& Jensen, J.. (2010). beta-Adrenoceptor stimulation potentiates insulin-stimulated PKB phosphorylation in rat cardiomyocytes via cAMP and PKA. in British Journal of Pharmacology, 160(1), 116-129. https://doi.org/10.1111/j.1476-5381.2010.00677.x
Stuenaes JT, Bolling A, Ingvaldsen A, Rommundstad C, Sudar E, Lin F, Lai Y, Jensen J. beta-Adrenoceptor stimulation potentiates insulin-stimulated PKB phosphorylation in rat cardiomyocytes via cAMP and PKA. in British Journal of Pharmacology. 2010;160(1):116-129. doi:10.1111/j.1476-5381.2010.00677.x .
Stuenaes, Jorid T., Bolling, Astrid, Ingvaldsen, Ada, Rommundstad, Camilla, Sudar, Emina, Lin, Fang-Chin, Lai, Yu-Chiang, Jensen, Jorgen, "beta-Adrenoceptor stimulation potentiates insulin-stimulated PKB phosphorylation in rat cardiomyocytes via cAMP and PKA" in British Journal of Pharmacology, 160, no. 1 (2010):116-129, https://doi.org/10.1111/j.1476-5381.2010.00677.x . .