Characterising the Inhibitory Actions of Ceramide upon Insulin Signaling in Different Skeletal Muscle Cell Models: A Mechanistic Insight

Ceramides are known to promote insulin resistance in a number of metabolically important tissues including skeletal muscle, the predominant site of insulin-stimulated glucose disposal. Depending on cell type, these lipid intermediates have been shown to inhibit protein kinase B (PKB/Akt), a key medi...

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Vydáno v:PloS one Ročník 9; číslo 7; s. e101865
Hlavní autoři: Mahfouz, Rana, Khoury, Rhéa, Blachnio-Zabielska, Agnieszka, Turban, Sophie, Loiseau, Nicolas, Lipina, Christopher, Stretton, Clare, Bourron, Olivier, Ferré, Pascal, Foufelle, Fabienne, Hundal, Harinder S., Hajduch, Eric
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States Public Library of Science 24.07.2014
Public Library of Science (PLoS)
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ISSN:1932-6203, 1932-6203
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Abstract Ceramides are known to promote insulin resistance in a number of metabolically important tissues including skeletal muscle, the predominant site of insulin-stimulated glucose disposal. Depending on cell type, these lipid intermediates have been shown to inhibit protein kinase B (PKB/Akt), a key mediator of the metabolic actions of insulin, via two distinct pathways: one involving the action of atypical protein kinase C (aPKC) isoforms, and the second dependent on protein phosphatase-2A (PP2A). The main aim of this study was to explore the mechanisms by which ceramide inhibits PKB/Akt in three different skeletal muscle-derived cell culture models; rat L6 myotubes, mouse C2C12 myotubes and primary human skeletal muscle cells. Our findings indicate that the mechanism by which ceramide acts to repress PKB/Akt is related to the myocellular abundance of caveolin-enriched domains (CEM) present at the plasma membrane. Here, we show that ceramide-enriched-CEMs are markedly more abundant in L6 myotubes compared to C2C12 myotubes, consistent with their previously reported role in coordinating aPKC-directed repression of PKB/Akt in L6 muscle cells. In contrast, a PP2A-dependent pathway predominantly mediates ceramide-induced inhibition of PKB/Akt in C2C12 myotubes. In addition, we demonstrate for the first time that ceramide engages an aPKC-dependent pathway to suppress insulin-induced PKB/Akt activation in palmitate-treated cultured human muscle cells as well as in muscle cells from diabetic patients. Collectively, this work identifies key mechanistic differences, which may be linked to variations in plasma membrane composition, underlying the insulin-desensitising effects of ceramide in different skeletal muscle cell models that are extensively used in signal transduction and metabolic studies.
AbstractList Ceramides are known to promote insulin resistance in a number of metabolically important tissues including skeletal muscle, the predominant site of insulin-stimulated glucose disposal. Depending on cell type, these lipid intermediates have been shown to inhibit protein kinase B (PKB/Akt), a key mediator of the metabolic actions of insulin, via two distinct pathways: one involving the action of atypical protein kinase C (aPKC) isoforms, and the second dependent on protein phosphatase-2A (PP2A). The main aim of this study was to explore the mechanisms by which ceramide inhibits PKB/Akt in three different skeletal muscle-derived cell culture models; rat L6 myotubes, mouse C2C12 myotubes and primary human skeletal muscle cells. Our findings indicate that the mechanism by which ceramide acts to repress PKB/Akt is related to the myocellular abundance of caveolin-enriched domains (CEM) present at the plasma membrane. Here, we show that ceramide-enriched-CEMs are markedly more abundant in L6 myotubes compared to C2C12 myotubes, consistent with their previously reported role in coordinating aPKC-directed repression of PKB/Akt in L6 muscle cells. In contrast, a PP2A-dependent pathway predominantly mediates ceramide-induced inhibition of PKB/Akt in C2C12 myotubes. In addition, we demonstrate for the first time that ceramide engages an aPKC-dependent pathway to suppress insulin-induced PKB/Akt activation in palmitate-treated cultured human muscle cells as well as in muscle cells from diabetic patients. Collectively, this work identifies key mechanistic differences, which may be linked to variations in plasma membrane composition, underlying the insulin-desensitising effects of ceramide in different skeletal muscle cell models that are extensively used in signal transduction and metabolic studies.
Ceramides are known to promote insulin resistance in a number of metabolically important tissues including skeletal muscle, the predominant site of insulin-stimulated glucose disposal. Depending on cell type, these lipid intermediates have been shown to inhibit protein kinase B (PKB/Akt), a key mediator of the metabolic actions of insulin, via two distinct pathways: one involving the action of atypical protein kinase C (aPKC) isoforms, and the second dependent on protein phosphatase-2A (PP2A). The main aim of this study was to explore the mechanisms by which ceramide inhibits PKB/Akt in three different skeletal muscle-derived cell culture models; rat L6 myotubes, mouse C2C12 myotubes and primary human skeletal muscle cells. Our findings indicate that the mechanism by which ceramide acts to repress PKB/Akt is related to the myocellular abundance of caveolin-enriched domains (CEM) present at the plasma membrane. Here, we show that ceramide-enriched-CEMs are markedly more abundant in L6 myotubes compared to C2C12 myotubes, consistent with their previously reported role in coordinating aPKC-directed repression of PKB/Akt in L6 muscle cells. In contrast, a PP2A-dependent pathway predominantly mediates ceramide-induced inhibition of PKB/Akt in C2C12 myotubes. In addition, we demonstrate for the first time that ceramide engages an aPKC-dependent pathway to suppress insulin-induced PKB/Akt activation in palmitate-treated cultured human muscle cells as well as in muscle cells from diabetic patients. Collectively, this work identifies key mechanistic differences, which may be linked to variations in plasma membrane composition, underlying the insulin-desensitising effects of ceramide in different skeletal muscle cell models that are extensively used in signal transduction and metabolic studies.Ceramides are known to promote insulin resistance in a number of metabolically important tissues including skeletal muscle, the predominant site of insulin-stimulated glucose disposal. Depending on cell type, these lipid intermediates have been shown to inhibit protein kinase B (PKB/Akt), a key mediator of the metabolic actions of insulin, via two distinct pathways: one involving the action of atypical protein kinase C (aPKC) isoforms, and the second dependent on protein phosphatase-2A (PP2A). The main aim of this study was to explore the mechanisms by which ceramide inhibits PKB/Akt in three different skeletal muscle-derived cell culture models; rat L6 myotubes, mouse C2C12 myotubes and primary human skeletal muscle cells. Our findings indicate that the mechanism by which ceramide acts to repress PKB/Akt is related to the myocellular abundance of caveolin-enriched domains (CEM) present at the plasma membrane. Here, we show that ceramide-enriched-CEMs are markedly more abundant in L6 myotubes compared to C2C12 myotubes, consistent with their previously reported role in coordinating aPKC-directed repression of PKB/Akt in L6 muscle cells. In contrast, a PP2A-dependent pathway predominantly mediates ceramide-induced inhibition of PKB/Akt in C2C12 myotubes. In addition, we demonstrate for the first time that ceramide engages an aPKC-dependent pathway to suppress insulin-induced PKB/Akt activation in palmitate-treated cultured human muscle cells as well as in muscle cells from diabetic patients. Collectively, this work identifies key mechanistic differences, which may be linked to variations in plasma membrane composition, underlying the insulin-desensitising effects of ceramide in different skeletal muscle cell models that are extensively used in signal transduction and metabolic studies.
Audience Academic
Author Ferré, Pascal
Loiseau, Nicolas
Blachnio-Zabielska, Agnieszka
Stretton, Clare
Foufelle, Fabienne
Khoury, Rhéa
Lipina, Christopher
Bourron, Olivier
Hundal, Harinder S.
Mahfouz, Rana
Hajduch, Eric
Turban, Sophie
AuthorAffiliation 2 Université Pierre et Marie Curie – Paris 6, UMR-S 1138, Paris, France
4 Department of Physiology, Medical University of Bialystok, Bialystok, Poland
5 Division of Cell Signalling and Immunology, College of Life Sciences, University of Dundee, Dundee, United Kingdom
6 INRA, UMR1331 Toxalim, Research Centre in Food Toxicology, Toulouse, France
3 Université Paris Descartes, UMR-S 1138, Paris, France
INSERM/UMR 1048, France
1 INSERM, UMR-S 1138, Centre de Recherche des Cordeliers, Paris, France
7 Département de Diabétologie et Maladies métaboliques, AP-HP, Hôpital Pitié-Salpêtrière, Paris, France
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ContentType Journal Article
Copyright COPYRIGHT 2014 Public Library of Science
2014 Mahfouz et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
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Conceived and designed the experiments: PF FF HSH EH. Performed the experiments: RM RK ST AB-Z NL CL CS. Analyzed the data: PF FF HSH EH. Contributed reagents/materials/analysis tools: OB. Contributed to the writing of the manuscript: CL ST FF EH.
Competing Interests: The authors have declared that no competing interests exist.
ORCID 0000-0003-0115-7045
0000-0002-0125-7751
0000-0003-3783-0879
0000-0002-0752-622X
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Snippet Ceramides are known to promote insulin resistance in a number of metabolically important tissues including skeletal muscle, the predominant site of...
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SubjectTerms AKT protein
Animal models
Animals
Biology and Life Sciences
Caveolin
Caveolins - genetics
Caveolins - metabolism
Cell culture
Cellular signal transduction
Ceramide
Ceramides
Ceramides - pharmacology
Composition effects
Diabetes mellitus
Gene Expression Regulation
Glucose
Human health and pathology
Humans
Insulin
Insulin - metabolism
Insulin - pharmacology
Insulin resistance
Intermediates
Isoforms
Kinases
Life Sciences
Membrane composition
Mice
Muscle Fibers, Skeletal - cytology
Muscle Fibers, Skeletal - drug effects
Muscle Fibers, Skeletal - metabolism
Muscle, Skeletal - cytology
Muscle, Skeletal - drug effects
Muscle, Skeletal - metabolism
Muscles
Myotubes
Palmitic acid
Phosphoprotein phosphatase
Plant lipids
Primary Cell Culture
Protein kinase C
Protein Kinase C - genetics
Protein Kinase C - metabolism
Protein phosphatase
Protein Phosphatase 2 - genetics
Protein Phosphatase 2 - metabolism
Proto-Oncogene Proteins c-akt - genetics
Proto-Oncogene Proteins c-akt - metabolism
Rats
Rodents
Signal transduction
Signal Transduction - drug effects
Signaling
Skeletal muscle
Tissues
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Title Characterising the Inhibitory Actions of Ceramide upon Insulin Signaling in Different Skeletal Muscle Cell Models: A Mechanistic Insight
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