Ablation of triadin causes loss of cardiac Ca2+ release units, impaired excitation-contraction coupling, and cardiac arrhythmias

Division of Clinical Pharmacology, Department of Medicine, Vanderbilt University, Nashville, TN 37232, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 05/2009; 106(18):7636-41. DOI: 10.1073/pnas.0902919106
Source: PubMed

ABSTRACT Heart muscle excitation-contraction (E-C) coupling is governed by Ca(2+) release units (CRUs) whereby Ca(2+) influx via L-type Ca(2+) channels (Cav1.2) triggers Ca(2+) release from juxtaposed Ca(2+) release channels (RyR2) located in junctional sarcoplasmic reticulum (jSR). Although studies suggest that the jSR protein triadin anchors cardiac calsequestrin (Casq2) to RyR2, its contribution to E-C coupling remains unclear. Here, we identify the role of triadin using mice with ablation of the Trdn gene (Trdn(-/-)). The structure and protein composition of the cardiac CRU is significantly altered in Trdn(-/-) hearts. jSR proteins (RyR2, Casq2, junctin, and junctophilin 1 and 2) are significantly reduced in Trdn(-/-) hearts, whereas Cav1.2 and SERCA2a remain unchanged. Electron microscopy shows fragmentation and an overall 50% reduction in the contacts between jSR and T-tubules. Immunolabeling experiments show reduced colocalization of Cav1.2 with RyR2 and substantial Casq2 labeling outside of the jSR in Trdn(-/-) myocytes. CRU function is impaired in Trdn(-/-) myocytes, with reduced SR Ca(2+) release and impaired negative feedback of SR Ca(2+) release on Cav1.2 Ca(2+) currents (I(Ca)). Uninhibited Ca(2+) influx via I(Ca) likely contributes to Ca(2+) overload and results in spontaneous SR Ca(2+) releases upon beta-adrenergic receptor stimulation with isoproterenol in Trdn(-/-) myocytes, and ventricular arrhythmias in Trdn(-/-) mice. We conclude that triadin is critically important for maintaining the structural and functional integrity of the cardiac CRU; triadin loss and the resulting alterations in CRU structure and protein composition impairs E-C coupling and renders hearts susceptible to ventricular arrhythmias.

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Available from: Clara Franzini-Armstrong, Feb 03, 2015
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    • "In contrast, the frequency and extent of JMCs are largely preserved in Casq2-null cardiomyocytes, although SR cisternae are significantly enlarged in these cells [21]. In Trdnnull mice, the number and the size of dyadic junctions are reduced in cardiomyocytes [22], while triad alteration is subtler in skeletal muscles [23]. Specifically, triad orientation is modified and calsequestrin is occasionally mis-localized in triadin-deficient skeletal muscle cells [23]. "
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    ABSTRACT: Excitable cells typically possess junctional membrane complexes (JMCs) constructed by the plasma membrane and the endo/sarcoplasmic reticulum (ER/SR) for channel crosstalk. These JMCs are termed triads in skeletal muscle, dyads in cardiac muscle, peripheral couplings in smooth and developing striated muscles, and subsurface cisterns in neurons. Junctophilin subtypes contribute to the formation and maintenance of JMCs by serving as a physical bridge between the plasma membrane and ER/SR membrane in different cell types. In muscle cells, junctophilin deficiency prevents JMC formation and functional crosstalk between cell-surface Ca2+ channels and ER/SR Ca2+ release channels. Human genetic mutations in junctophilin subtypes are linked to congenital hypertrophic cardiomyopathy and neurodegenerative diseases. Furthermore, growing evidence suggests that dysregulation of junctophilins induces pathological alterations in skeletal and cardiac muscle.
    Cell Calcium 01/2015; 58(4). DOI:10.1016/j.ceca.2015.01.007 · 3.51 Impact Factor
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    • "Thereby, lowering the SR Ca 2+ concentration during a spark would make the RyRs insensitive for Ca 2+ on the cytosolic side of the channels, which causes their deactivation. Based on observations in SR vesicles, RyRs in lipid bilayers and cells overexpressing calsequestrin, deactivation has been suggested to occur via a retrograde signal mediated by allosteric interactions between calsequestrin (acting as the Ca 2+ sensor) and junctin and/or triadin and the RyR [2] [8] [49] [50]. This mode of spark termination could be stabilized by a reinforcing mechanism that has been proposed recently based on model predictions. "
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    ABSTRACT: In cardiac muscle, a number of posttranslational protein modifications can alter the function of the Ca(2+) release channel of the sarcoplasmic reticulum (SR), also known as the ryanodine receptor (RyR). During every heartbeat RyRs are activated by the Ca(2+)-induced Ca(2+) release mechanism and contribute a large fraction of the Ca(2+) required for contraction. Some of the posttranslational modifications of the RyR are known to affect its gating and Ca(2+) sensitivity. Presently, research in a number of laboratories is focused on RyR phosphorylation, both by PKA and CaMKII, or on RyR modifications caused by reactive oxygen and nitrogen species (ROS/RNS). Both classes of posttranslational modifications are thought to play important roles in the physiological regulation of channel activity, but are also known to provoke abnormal alterations during various diseases. Only recently it was realized that several types of posttranslational modifications are tightly connected and form synergistic (or antagonistic) feed-back loops resulting in additive and potentially detrimental downstream effects. This review summarizes recent findings on such posttranslational modifications, attempts to bridge molecular with cellular findings, and opens a perspective for future work trying to understand the ramifications of crosstalk in these multiple signaling pathways. Clarifying these complex interactions will be important in the development of novel therapeutic approaches, since this may form the foundation for the implementation of multi-pronged treatment regimes in the future. This article is part of a Special Issue entitled: Cardiomyocyte Biology: Cardiac Pathways of Differentiation, Metabolism and Contraction.
    Biochimica et Biophysica Acta 08/2012; 1833(4). DOI:10.1016/j.bbamcr.2012.08.016 · 4.66 Impact Factor
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    • "Rather, Trisk 95 is deemed to facilitate the orthograde signal between the DHPR and RyR1 in excitation-contraction coupling (EC-coupling) [18]. Results from siRNA-mediated triadin knockdown [19] and pan-triadin-knockout studies [20] support these findings. It should be noted that triadin knockout mice show normal skeletal contractility, which may be due to a contribution from numerous compensatory mechanisms [21]. "
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    ABSTRACT: Excitation-contraction coupling in skeletal muscle depends, in part, on a functional interaction between the ligand-gated ryanodine receptor (RyR1) and integral membrane protein Trisk 95, localized to the sarcoplasmic reticulum membrane. Various domains on Trisk 95 can associate with RyR1, yet the domain responsible for regulating RyR1 activity has remained elusive. We explored the hypothesis that a luminal Trisk 95 KEKE motif (residues 200-232), known to promote RyR1 binding, may also form the RyR1 activation domain. Peptides corresponding to Trisk 95 residues 200-232 or 200-231 bound to RyR1 and increased the single channel activity of RyR1 by 1.49 ± 0.11-fold and 1.8 ± 0.15-fold respectively, when added to its luminal side. A similar increase in [(3)H]ryanodine binding, which reflects open probability of the channels, was also observed. This RyR1 activation is similar to activation induced by full length Trisk 95. Circular dichroism showed that both peptides were intrinsically disordered, suggesting a defined secondary structure is not necessary to mediate RyR1 activation. These data for the first time demonstrate that Trisk 95's 200-231 region is responsible for RyR1 activation. Furthermore, it shows that no secondary structure is required to achieve this activation, the Trisk 95 residues themselves are critical for the Trisk 95-RyR1 interaction.
    PLoS ONE 08/2012; 7(8):e43817. DOI:10.1371/journal.pone.0043817 · 3.23 Impact Factor
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