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01-11-2014 | Original Contribution

Ceramide-mediated depression in cardiomyocyte contractility through PKC activation and modulation of myofilament protein phosphorylation

Authors: Jillian N. Simon, Shamim A. K. Chowdhury, Chad M. Warren, Sakthivel Sadayappan, David F. Wieczorek, R. John Solaro, Beata M. Wolska

Published in: Basic Research in Cardiology | Issue 6/2014

Abstract

Although ceramide accumulation in the heart is considered a major factor in promoting apoptosis and cardiac disorders, including heart failure, lipotoxicity and ischemia–reperfusion injury, little is known about ceramide’s role in mediating changes in contractility. In the present study, we measured the functional consequences of acute exposure of isolated field-stimulated adult rat cardiomyocytes to C₆-ceramide. Exogenous ceramide treatment depressed the peak amplitude and the maximal velocity of shortening without altering intracellular calcium levels or kinetics. The inactive ceramide analog C₆-dihydroceramide had no effect on myocyte shortening or [Ca²⁺]_i transients. Experiments testing a potential role for C₆-ceramide-mediated effects on activation of protein kinase C (PKC) demonstrated evidence for signaling through the calcium-independent isoform, PKCε. We employed 2-dimensional electrophoresis and anti-phospho-peptide antibodies to test whether treatment of the cardiomyocytes with C₆-ceramide altered myocyte shortening via PKC-dependent phosphorylation of myofilament proteins. Compared to controls, myocytes treated with ceramide exhibited increased phosphorylation of myosin binding protein-C (cMyBP-C), specifically at Ser273 and Ser302, and troponin I (cTnI) at sites apart from Ser23/24, which could be attenuated with PKC inhibition. We conclude that the altered myofilament response to calcium resulting from multiple sites of PKC-dependent phosphorylation contributes to contractile dysfunction that is associated with cardiac diseases in which elevations in ceramides are present.

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Title: Ceramide-mediated depression in cardiomyocyte contractility through PKC activation and modulation of myofilament protein phosphorylation
Authors: Jillian N. Simon
Shamim A. K. Chowdhury
Chad M. Warren
Sakthivel Sadayappan
David F. Wieczorek
R. John Solaro
Beata M. Wolska
Publication date: 01-11-2014
Publisher: Springer Berlin Heidelberg
Published in: Basic Research in Cardiology / Issue 6/2014
Print ISSN: 0300-8428
Electronic ISSN: 1435-1803
DOI: https://doi.org/10.1007/s00395-014-0445-6

2024 ASCO Annual Meeting

Springer Medicine

Ceramide-mediated depression in cardiomyocyte contractility through PKC activation and modulation of myofilament protein phosphorylation

Abstract

2024 ASCO Annual Meeting

Springer Medicine

Abstract

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