Characterising the inhibitory actions of ceramide upon insulin signaling in different skeletal muscle cell models: a mechanistic insight

Rana Mahfouz, Rhéa Khoury, Agnieszka Blachnio-Zabielska, Sophie Turban, Nicolas Loiseau, Christopher Lipina, Clare Stretton, Olivier Bourron, Pascal Ferré, Fabienne Foufelle, Harinder S. Hundal, Eric Hajduch (Lead / Corresponding author)

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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.
Original languageEnglish
Article numbere101865
Number of pages9
JournalPLoS ONE
Volume9
Issue number7
DOIs
Publication statusPublished - 24 Jul 2014

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ceramides
Ceramides
mechanistic models
myocytes
Muscle Cells
Muscle
skeletal muscle
Skeletal Muscle
Skeletal Muscle Fibers
insulin
Cells
Insulin
protein kinase C
Protein Phosphatase 2
phosphoprotein phosphatase
Cell membranes
plasma membrane
Cell Membrane
Caveolins
Signal transduction

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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. / Characterising the inhibitory actions of ceramide upon insulin signaling in different skeletal muscle cell models : a mechanistic insight. In: PLoS ONE. 2014 ; Vol. 9, No. 7.
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Mahfouz, R, Khoury, R, Blachnio-Zabielska, A, Turban, S, Loiseau, N, Lipina, C, Stretton, C, Bourron, O, Ferré, P, Foufelle, F, Hundal, HS & Hajduch, E 2014, 'Characterising the inhibitory actions of ceramide upon insulin signaling in different skeletal muscle cell models: a mechanistic insight', PLoS ONE, vol. 9, no. 7, e101865. https://doi.org/10.1371/journal.pone.0101865

Characterising the inhibitory actions of ceramide upon insulin signaling in different skeletal muscle cell models : a mechanistic insight. / 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 (Lead / Corresponding author).

In: PLoS ONE, Vol. 9, No. 7, e101865, 24.07.2014.

Research output: Contribution to journalArticle

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T1 - Characterising the inhibitory actions of ceramide upon insulin signaling in different skeletal muscle cell models

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AU - Mahfouz, Rana

AU - Khoury, Rhéa

AU - Blachnio-Zabielska, Agnieszka

AU - Turban, Sophie

AU - Loiseau, Nicolas

AU - Lipina, Christopher

AU - Stretton, Clare

AU - Bourron, Olivier

AU - Ferré, Pascal

AU - Foufelle, Fabienne

AU - Hundal, Harinder S.

AU - Hajduch, Eric

PY - 2014/7/24

Y1 - 2014/7/24

N2 - 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.

AB - 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.

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