Store-operated calcium entry contributes to abnormal Ca2+ signalling in dystrophic mdx mouse myoblasts

Onopiuk, Marta; Brutkowski, Wojciech; Young, Chris; Krasowska, Elżbieta; Róg, Justyna; Ritso, Morten; Wojciechowska, Sylwia; Arkle, S.; Zabłocki, Krzysztof; Górecki, Dariusz C.

doi:10.1016/j.abb.2015.01.025

Cited by 30 publications

(34 citation statements)

References 49 publications

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“…All findings here together with those published previously [15,22] clearly indicate that calcium signalling in mdx myoblasts and w/t cells varies and the amplitude of calcium response is always higher in the mdx myoblasts than in their w/t equivalents. In this study mice myoblasts were stimulated with ATP or UTP under calcium-free conditions to avoid any P2X-dependent calcium entry (in a case of ATP as a stimulus) and an activation of the storeoperated calcium entry.…”

Section: Discussionsupporting

confidence: 87%

“…Besides the aberrant ionotropic response to extracellular ATP, we have also found increased activity of store-operated calcium entry mechanisms in dystrophic myoblasts [22]. These and other studies demonstrate that abnormalities of calcium homeostasis in DMD occur in myoblasts and not only in fully differentiated myofibres and that they involve alterations of specific cellular mechanisms rather than mechanical membrane instability.…”

Section: Introductionsupporting

confidence: 77%

“…Moreover, prolonged stimulation of mdx myoblasts with ATP or more selectively with P2RX7-specific agonist BzATP led to the opening of a large pore and additional effects such as stimulated autophagy and MMP-2 release [18,45]. In these cells as well as in the primary mdx myoblasts an increased level of STIM1 protein seems to be behind significantly increased store-operated calcium entry (SOCE) [22 ]. In this paper the first step of cellular response to a stimulation of metabotropic nucleotide receptors, leading to a depletion of ER stores which is prerequisite for SOCE activation has been investigated.…”

Section: Discussionmentioning

confidence: 99%

“…These include: (i) Ca 2+ influx through P2RX channels and (ii) store-operated calcium entry (SOCE), which both allow extracellular Ca 2+ to flow into the cell. Both mechanisms have been identified and analysed by us previously in immortalized and primary w/t and mdx myoblasts [15,22]. The third mechanism concerning the eATP-evoked response is Ca 2+ release from the ER stores due to stimulation of metabotropic nucleotide receptors of the P2RY family.…”

Section: Atp Sensitive Nucleotide Receptorsmentioning

confidence: 98%

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Dystrophic mdx mouse myoblasts exhibit elevated ATP/UTP-evoked metabotropic purinergic responses and alterations in calcium signalling

Róg

Oksiejuk

Gosselin

et al. 2019

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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Running title: mdx mutation affects calcium signalling in myoblastsHighlights:1. NGS reveals changes in Ca 2+ signalling-related RNAs in mdx myoblasts 2. Mdx myoblasts exerts increased susceptibility to P2RY2-mediated stimulation 3. Levels of several Ca 2+ signalling-related proteins are changed in mdx myoblasts 4. P2RY2 agonist slows-down mdx myoblasts motility SummaryPathophysiology of Duchenne Muscular Dystrophy is still elusive. Although progressive damage to muscle fibres is a cause of muscle deterioration leading to premature death, there is a growing body of evidence indicating that the triggering effects of DMD mutation are present at the very early stage of muscle development. Previously, elevated activity of P2X7 receptors and increased store-operated calcium entry were shown in myoblasts derived from mdx mice. Here, the metabotropic extracellular ATP/UTP-evoked response has been investigated. Sensitivity to antagonist, effect of gene silencing and cellular localization studies showed that elevated purinergic responses in mdx myoblasts have been caused by increased expression of P2Y2 but not P2Y4 receptors. These alterations have physiological implications as shown by reduced motility of mdx myoblasts upon treatment with P2Y2 agonist. The ultimate increase in intracellular calcium in dystrophic cells reflected complex alterations of calcium homeostasis identified in the RNA seq data and with significant modulation at the protein level including a decrease of Gq11 subunit , PMCA, IP3-receptor and elevation of PLC, SERCA and NCX. In conclusion, whereas specificity of dystrophic myoblast excitation by extracellular ATP is determined by specific receptor overexpression, the intensity of this altered response depends on relative activities of downstream calcium regulators that also accompany the Dmd gene mutation. These results confirm that phenotypic effects of DMD emerge in undifferentiated muscle cells and not only due to the absence of dystrophin protein in myofibres. Therefore, the pathogenesis of DMD and the relevance of current therapeutic approaches may need re-evaluation.

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Section: Discussionsupporting

confidence: 87%

Section: Introductionsupporting

confidence: 77%

Section: Discussionmentioning

confidence: 99%

Section: Atp Sensitive Nucleotide Receptorsmentioning

confidence: 98%

See 2 more Smart Citations

Dystrophic mdx mouse myoblasts exhibit elevated ATP/UTP-evoked metabotropic purinergic responses and alterations in calcium signalling

Róg

Oksiejuk

Gosselin

et al. 2019

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

Self Cite

View full text Add to dashboard Cite

show abstract

“…Mutations in the STIM1 gene also are associated with SCID and also are accompanied by skeletal muscle atrophy and myopathy due to a reduction in SOCE [33,56]. Myotubes from mdx mouse models show higher expression of STIM1 with changes in other Ca 2+ -handling proteins [57,58]. A muscular hypotonia-associated STIM1 mutation at R429 induces abnormalities in intracellular Ca 2+ movement, mitochondria and SOCE in mouse skeletal myotubes [59].…”

Section: Introductionmentioning

confidence: 99%

TRPCs: Influential Mediators in Skeletal Muscle

Choi

Jeong

et al. 2020

Cells

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Ca 2+ itself or Ca 2+ -dependent signaling pathways play fundamental roles in various cellular processes from cell growth to death. The most representative example can be found in skeletal muscle cells where a well-timed and adequate supply of Ca 2+ is required for coordinated Ca 2+ -dependent skeletal muscle functions, such as the interactions of contractile proteins during contraction. Intracellular Ca 2+ movements between the cytosol and sarcoplasmic reticulum (SR) are strictly regulated to maintain the appropriate Ca 2+ supply in skeletal muscle cells. Added to intracellular Ca 2+ movements, the contribution of extracellular Ca 2+ entry to skeletal muscle functions and its significance have been continuously studied since the early 1990s. Here, studies on the roles of channel proteins that mediate extracellular Ca 2+ entry into skeletal muscle cells using skeletal myoblasts, myotubes, fibers, tissue, or skeletal muscle-originated cell lines are reviewed with special attention to the proposed functions of transient receptor potential canonical proteins (TRPCs) as store-operated Ca 2+ entry (SOCE) channels under normal conditions and the potential abnormal properties of TRPCs in muscle diseases such as Duchenne muscular dystrophy (DMD).

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Investigating the Involvement of C−X−C Motif Chemokine 5 and P2X7 Purinoceptor in Ectopic Calcification in Mouse Models of Duchenne Muscular Dystrophy

Rumney,

Pomeroy,

Górecki

2024

J of Cellular Biochemistry

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Ectopic calcification of myofibers is an early pathogenic feature in patients and animal models of Duchenne muscular dystrophy (DMD). In previous studies using the Dmdmdx‐βgeo mouse model, we found that the dystrophin‐null phenotype exacerbates this abnormality and that mineralised myofibers are surrounded by macrophages. Furthermore, the P2X7 purinoceptor, functioning in immune cells offers protection against dystrophic calcification. In the present study, by exploring transcriptomic data from Dmdmdx mice, we hypothesised these effects to be mediated by C−X−C motif chemokine 5 (CXCL5) downstream of P2X7 activation. We found that CXCL5 is upregulated in the quadriceps muscles of Dmdmdx‐βgeo mice compared to wild‐type controls. In contrast, at the cell level, dystrophic (SC5) skeletal muscle cells secreted less CXCL5 chemokine than wild‐type (IMO) controls. Although release from IMO cells was increased by P2X7 activation, this could not explain the elevated CXCL5 levels observed in dystrophic muscle tissue. Instead, we found that CXCL5 is released by dystrophin‐null macrophages in response to P2X7 activation, suggesting that macrophages are the source of CXCL5 in dystrophic muscles. The effects of CXCL5 upon mineralisation were investigated using the Alizarin Red assay to quantify calcium deposition in vitro. In basal (low phosphate) media, CXCL5 increased calcification in IMO but not SC5 myoblasts. However, in cultures treated in high phosphate media, to mimic dysregulated phosphate metabolism occurring in DMD, CXCL5 decreased calcification in both IMO and SC5 cells. These data indicate that CXCL5 is part of a homoeostatic mechanism regulating intracellular calcium, that CXCL5 can be released by macrophages in response to the extracellular ATP damage‐associated signal, and that CXCL5 can be part of a damage response to protect against ectopic calcification. This mechanism is affected by DMD gene mutations.

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Store-operated calcium entry contributes to abnormal Ca2+ signalling in dystrophic mdx mouse myoblasts

Cited by 30 publications

References 49 publications

Dystrophic mdx mouse myoblasts exhibit elevated ATP/UTP-evoked metabotropic purinergic responses and alterations in calcium signalling

Dystrophic mdx mouse myoblasts exhibit elevated ATP/UTP-evoked metabotropic purinergic responses and alterations in calcium signalling

TRPCs: Influential Mediators in Skeletal Muscle

Investigating the Involvement of C−X−C Motif Chemokine 5 and P2X7 Purinoceptor in Ectopic Calcification in Mouse Models of Duchenne Muscular Dystrophy

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