Featured Article: AMPKα2 deficiency enhanced susceptibility to ventricular arrhythmias in mice by the role of β-adrenoceptor signaling

AMP-activated protein kinase-α2 is the main catalytic subunit of the heart, which is mainly located in cardiac myocytes. The effect of AMP-activated protein kinase-α2 on the cardiac electrophysiology is barely studied. From the previous study, it is possible that AMP-activated protein kinase-α2 may have some effect on the electrophysiology of the heart. To prove the hypothesis, we used the AMP-activated protein kinase-α2 knockout (AMPKα2−/−) mice to estimate the electrophysiological characteristics of AMPKα2−/− mice and try to find the mechanism between them. We used AMP-activated protein kinase-α2 gene knockout (AMPKα2−/−) mice and control wild-type mice as the experimental animals. In the experiment, we measured the monophasic action potential duration and test the inducibility to ventricular arrhythmia in isolated mice heart with and without β-adrenoceptor antagonist metoprolol. Meanwhile, plasma concentration of catecholamine was collected. We found that AMPKα2−/− significantly shortened 90% repolarization of monophasic action potential (MAP) (MAPD90) than wild-type (47.4 ± 2.6 ms vs. 55.5 ± 2.4 ms, n = 10, P < 0.05) and were more vulnerable to be induced to ventricular arrhythmias (70% (7/10) vs. 10% (1/10), P < 0.05), accompanied by the higher concentration of catecholamine (epinephrine: 1.75 ± 0.18 nmol/L vs. 0.68 ± 0.10 nmol/L n = 10, P < 0.05; norepinephrine: 9.56 ± 0.71 nmol/L vs. 2.52 ± 0.31 nmol/L n = 10, P < 0.05). The shortening of MAPD90 and increased inducibility to ventricular arrhythmias of AMPKα2−/− could almost be abolished when perfusion with β-adrenoceptor antagonist metoprolol. It indicated that the β-adrenoceptor activation resulting from catecholamine release was mainly responsible for the relating changes of electrophysiology of AMPKα2−/−. It had great clinical significance, as in patients who had problem with AMP-activated protein kinase-α2 gene, we might use β-adrenoceptor antagonists as the prevention of arrhythmias in future. Impact statement As far as we know, this is the first time the role of AMP-activated protein kinase-α2 (AMPKα2) on the cardiac electrophysiology is explored, and we found that the β-adrenoceptor activation resulting from catecholamine release was mainly responsible for the changes of electrophysiology related to the absence of AMPKα2. This has great clinical significance, as in patients who have problems with AMPKα2 gene, we may use β-adrenoceptor antagonists for the prevention of arrhythmias in future.

Download Full-text

Role of Adenosine 5′-Monophosphate-Activated Protein Kinase in Interleukin-6 Release from Isolated Mouse Skeletal Muscle

Endocrinology ◽

10.1210/en.2008-1204 ◽

2009 ◽

Vol 150 (2) ◽

pp. 600-606 ◽

Cited By ~ 26

Author(s):

Stephan Glund ◽

Jonas T. Treebak ◽

Yun Chau Long ◽

Romain Barres ◽

Benoit Viollet ◽

...

Keyword(s):

Skeletal Muscle ◽

Protein Kinase ◽

Extensor Digitorum Longus ◽

Protein Release ◽

Extensor Digitorum ◽

Whole Body ◽

Wild Type ◽

Amp Activated Protein Kinase ◽

Respective Wild Type

IL-6 is released from skeletal muscle during exercise and has consequently been implicated to mediate beneficial effects on whole-body metabolism. Using 5-aminoimidazole-4-carboxamide-1-β-4-ribofuranoside (AICAR), a pharmacological activator of 5′-AMP-activated protein kinase (AMPK), we tested the hypothesis that AMPK modulates IL-6 release from isolated muscle. Skeletal muscle from AMPKα2 kinase-dead transgenic, AMPKα1 knockout (KO) and AMPKγ3 KO mice and respective wild-type littermates was incubated in vitro, in the absence or presence of 2 mmol/liter AICAR. Skeletal muscle from wild-type mice was also incubated with the AMPK activator A-769662. Incubation of mouse glycolytic extensor digitorum longus and oxidative soleus muscle for 2 h was associated with profound IL-6 mRNA production and protein release, which was suppressed by AICAR (P < 0.001). Basal IL-6 release from soleus was increased between AMPKα2 kinase-dead and AMPKα1 KO and their respective wild-type littermates (P < 0.05), suggesting AMPK participates in the regulation of IL-6 release from oxidative muscle. The effect of AICAR on muscle IL-6 release was similar between AMPKα2 KD, AMPKα1 KO, and AMPKγ3 KO mice and their respective wild-type littermates (P < 0.001), indicating AICAR-mediated suppression of IL-6 mRNA expression and protein release is independent of AMPK function. However, IL-6 release from soleus, but not extensor digitorum longus, was reduced 45% by A-769662. Our results on basal and A-769662-mediated IL-6 release provide evidence for a role of AMPK in the regulation of IL-6 release from oxidative skeletal muscle. Furthermore, in addition to activating AMPK, AICAR suppresses IL-6 release by an unknown, AMPK-independent mechanism. Using transgenic and knockout mouse models to perturb AMP-activated protein kinase (AMPK) signaling, we provide evidence that AMPK-dependent pathways regulate IL-6 release from isolated oxidative skeletal muscle.

Download Full-text

The regulation of AMP-activated protein kinase by phosphorylation

Biochemical Journal ◽

10.1042/bj3450437 ◽

2000 ◽

Vol 345 (3) ◽

pp. 437-443 ◽

Cited By ~ 270

Author(s):

Silvie C. STEIN ◽

Angela WOODS ◽

Neil A. JONES ◽

Matthew D. DAVISON ◽

David CARLING

Keyword(s):

Protein Kinase ◽

Site Directed Mutagenesis ◽

Wild Type ◽

Aspartic Acid Residue ◽

Α Subunit ◽

Ampk Activity ◽

Amp Activated Protein Kinase ◽

Mutant Complex

The AMP-activated protein kinase (AMPK) cascade is activated by an increase in the AMP/ATP ratio within the cell. AMPK is regulated allosterically by AMP and by reversible phosphorylation. Threonine-172 within the catalytic subunit (α) of AMPK (Thr172) was identified as the major site phosphorylated by the AMP-activated protein kinase kinase (AMPKK) in vitro. We have used site-directed mutagenesis to study the role of phosphorylation of Thr172 on AMPK activity. Mutation of Thr172 to an aspartic acid residue (T172D) in either α1 or α2 resulted in a kinase complex with approx. 50% the activity of the corresponding wild-type complex. The activity of wild-type AMPK decreased by greater than 90% following treatment with protein phosphatases, whereas the activity of the T172D mutant complex fell by only 10-15%. Mutation of Thr172 to an alanine residue (T172A) almost completely abolished kinase activity. These results indicate that phosphorylation of Thr172 accounts for most of the activation by AMPKK, but that other sites are involved. In support of this we have shown that AMPKK phosphorylates at least two other sites on the α subunit and one site on the β subunit. Furthermore, we provide evidence that phosphorylation of Thr172 may be involved in the sensitivity of the AMPK complex to AMP.

Download Full-text

Faculty Opinions recommendation of Evaluation of the role of AMP-activated protein kinase and its downstream targets in mammalian hibernation.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1022352.346456 ◽

2005 ◽

Author(s):

Bruce Kemp

Keyword(s):

Protein Kinase ◽

Downstream Targets ◽

Amp Activated Protein Kinase ◽

Mammalian Hibernation

Download Full-text

The role of AMP-activated protein kinase in quercetin-induced apoptosis of HL-60 cells

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/gmu014 ◽

2014 ◽

Vol 46 (5) ◽

pp. 394-400 ◽

Cited By ~ 5

Author(s):

J. Xiao ◽

G. Niu ◽

S. Yin ◽

S. Xie ◽

Y. Li ◽

...

Keyword(s):

Protein Kinase ◽

Induced Apoptosis ◽

Amp Activated Protein Kinase

Download Full-text

Role of the AMP-activated protein kinase in regulating fatty acid metabolism during exerciseThis paper is one of a selection of papers published in this Special Issue, entitled 14th International Biochemistry of Exercise Conference – Muscles as Molecular and Metabolic Machines, and has undergone the Journal’s usual peer review process.

Applied Physiology Nutrition and Metabolism ◽

10.1139/h09-009 ◽

2009 ◽

Vol 34 (3) ◽

pp. 315-322 ◽

Cited By ~ 26

Author(s):

Gregory R. Steinberg

Keyword(s):

Skeletal Muscle ◽

Fatty Acids ◽

Adipose Tissue ◽

Fatty Acid ◽

Protein Kinase ◽

Fatty Acid Metabolism ◽

Acid Metabolism ◽

Moderate Intensity ◽

Amp Activated Protein Kinase

During moderate-intensity exercise, fatty acids are the predominant substrate for working skeletal muscle. The release of fatty acids from adipose tissue stores, combined with the ability of skeletal muscle to actively fine tune the gradient between fatty acid and carbohydrate metabolism, depending on substrate availability and energetic demands, requires a coordinated system of metabolic control. Over the past decade, since the discovery that AMP-activated protein kinase (AMPK) was increased in accordance with exercise intensity, there has been significant interest in the proposed role of this ancient stress-sensing kinase as a critical integrative switch controlling metabolic responses during exercise. In this review, studies examining the role of AMPK as a regulator of fatty acid metabolism in both adipose tissue and skeletal muscle during exercise will be discussed. Exercise induces activation of AMPK in adipocytes and regulates triglyceride hydrolysis and esterfication through phosphorylation of hormone sensitive lipase (HSL) and glycerol-3-phosphate acyl-transferase, respectively. In skeletal muscle, exercise-induced activation of AMPK is associated with increases in fatty acid uptake, phosphorylation of HSL, and increased fatty acid oxidation, which is thought to occur via the acetyl-CoA carboxylase-malony-CoA-CPT-1 signalling axis. Despite the importance of AMPK in regulating fatty acid metabolism under resting conditions, recent evidence from transgenic models of AMPK deficiency suggest that alternative signalling pathways may also be important for the control of fatty acid metabolism during exercise.

Download Full-text

AMP kinase is not required for the GLUT4 response to exercise and denervation in skeletal muscle

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00080.2004 ◽

2004 ◽

Vol 287 (4) ◽

pp. E739-E743 ◽

Cited By ~ 46

Author(s):

Burton F. Holmes ◽

David B. Lang ◽

Morris J. Birnbaum ◽

James Mu ◽

G. Lynis Dohm

Keyword(s):

Skeletal Muscle ◽

Dominant Negative ◽

Treadmill Running ◽

Wild Type ◽

Α Subunit ◽

Ampk Activity ◽

Denervated Muscles ◽

Amp Activated Protein Kinase ◽

Response To Exercise

An acute bout of exercise increases muscle GLUT4 mRNA in mice, and denervation decreases GLUT4 mRNA. AMP-activated protein kinase (AMPK) activity in skeletal muscle is also increased by exercise, and GLUT4 mRNA is increased in mouse skeletal muscle after treatment with AMPK activator 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside(AICAR). These findings suggest that AMPK activation might be responsible for the increase in GLUT4 mRNA expression in response to exercise. To investigate the role of AMPK in GLUT4 regulation in response to exercise and denervation, transgenic mice with a mutated AMPK α-subunit (dominant negative; AMPK-DN) were studied. GLUT4 did not increase in AMPK-DN mice that were treated with AICAR, demonstrating that muscle AMPK is inactive. Exercise (two 3-h bouts of treadmill running separated by 1 h of rest) increased GLUT4 mRNA in both wild-type and AMPK-DN mice. Likewise, denervation decreased GLUT4 mRNA in both wild-type and AMPK-DN mice. GLUT4 mRNA was also increased by AICAR treatment in both the innervated and denervated muscles. These data demonstrate that AMPK is not required for the response of GLUT4 mRNA to exercise and denervation.

Download Full-text

Targeting of Protein Kinase C-ϵ during Fcγ Receptor-dependent Phagocytosis Requires the ϵC1B Domain and Phospholipase C-γ1

Molecular Biology of the Cell ◽

10.1091/mbc.e04-12-1100 ◽

2006 ◽

Vol 17 (2) ◽

pp. 799-813 ◽

Cited By ~ 35

Author(s):

Keylon L. Cheeseman ◽

Takehiko Ueyama ◽

Tanya M. Michaud ◽

Kaori Kashiwagi ◽

Demin Wang ◽

...

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Phospholipase C ◽

Phospholipase D ◽

Fluorescent Protein ◽

Wild Type ◽

Fcγ Receptor ◽

Kinase C ◽

Green Fluorescent

Protein kinase C-ϵ (PKC-ϵ) translocates to phagosomes and promotes uptake of IgG-opsonized targets. To identify the regions responsible for this concentration, green fluorescent protein (GFP)-protein kinase C-ϵ mutants were tracked during phagocytosis and in response to exogenous lipids. Deletion of the diacylglycerol (DAG)-binding ϵC1 and ϵC1B domains, or the ϵC1B point mutant ϵC259G, decreased accumulation at phagosomes and membrane translocation in response to exogenous DAG. Quantitation of GFP revealed that ϵC259G, ϵC1, and ϵC1B accumulation at phagosomes was significantly less than that of intact PKC-ϵ. Also, the DAG antagonist 1-hexadecyl-2-acetyl glycerol (EI-150) blocked PKC-ϵ translocation. Thus, DAG binding to ϵC1B is necessary for PKC-ϵ translocation. The role of phospholipase D (PLD), phosphatidylinositol-specific phospholipase C (PI-PLC)-γ1, and PI-PLC-γ2 in PKC-ϵ accumulation was assessed. Although GFP-PLD2 localized to phagosomes and enhanced phagocytosis, PLD inhibition did not alter target ingestion or PKC-ϵ localization. In contrast, the PI-PLC inhibitor U73122 decreased both phagocytosis and PKC-ϵ accumulation. Although expression of PI-PLC-γ2 is higher than that of PI-PLC-γ1, PI-PLC-γ1 but not PI-PLC-γ2 consistently concentrated at phagosomes. Macrophages from PI-PLC-γ2-/-mice were similar to wild-type macrophages in their rate and extent of phagocytosis, their accumulation of PKC-ϵ at the phagosome, and their sensitivity to U73122. This implicates PI-PLC-γ1 as the enzyme that supports PKC-ϵ localization and phagocytosis. That PI-PLC-γ1 was transiently tyrosine phosphorylated in nascent phagosomes is consistent with this conclusion. Together, these results support a model in which PI-PLC-γ1 provides DAG that binds to ϵC1B, facilitating PKC-ϵ localization to phagosomes for efficient IgG-mediated phagocytosis.

Download Full-text