B-lymphocytes from malignant hyperthermia-susceptible patients have an increased sensitivity to skeletal muscle by ryanodine receptor activators.

Girard, Thierry; Cavagna, Dario; Padovan, Elisabetta; Spagnoli, Giulio C.; Urwyler, Albert; Zorzato, Francesco; Treves, Susan

doi:10.1016/s0021-9258(19)61047-1

Cited by 9 publications

(20 citation statements)

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“…Amplification conditions were 5 min at 95 • C, followed by 40 cycles of 30 s annealing at 56 • C (24F/R), 71 • C (71F/R) or 60 • C (101F/R), 60 s extension at 72 • C and 30 s denaturation at 94 • C, followed by a final extension for 3 min at 72 • C. The presence of the nucleotide substitutions was detected by restriction enzyme digestion using BstUI, HhaI or AccI. In order to demonstrate that the cDNA preparations used for PCR amplification were not contaminated by genomic DNA, a PCR amplification spanning exons 39-40 was carried out as described previously [28].…”

Section: Mutation Screeningmentioning

confidence: 99%

“…Changes in [Ca 2+ ] i of the lymphoblastoid cells were monitored with the fluorescent Ca 2+ indicator fura 2. Experiments were carried out on populations of cells, in a PerkinElmer LS-50 spectrofluorimeter as described previously [28,30,31] or at the single-cell level by digital imaging microscopy as described previously [32]. In the latter case, lymphoblastoid cells loaded with 5 µM fura 2 were allowed to attach to poly(L-lysine)treated glass coverslips for 10 min before the experiments.…”

Section: Intracellular Ca 2+ Measurementsmentioning

confidence: 99%

“…We first created dose-response curves to 4-cmc by studying changes in [Ca 2+ ] i in cell populations in Ca 2+ -free Krebs-Ringer solution and expressed these as percentage release with respect to the thapsigargin-sensitive pool (set as 100 %). 4-cmc is a specific activator of the RyR1 and can be used pharmacologically to discriminate between MHN (MH-normal) and MHS (MHsusceptible) [33][34][35]; when RYR1 mutations are associated with the MHS phenotype, the sensitivity of the receptor to 4-cmc is increased [28,34]. Figure 4 shows the dose-response curves to 4-cmc in lymphoblastoid cells from control individuals, from individuals harbouring the R999H substitution and from individuals harbouring the P3527S and V4849I substitutions in the homo- zygous and heterozygous states.…”

Section: Sensitivity Of Lymphoblastoid Cell Lines To Pharmacological Activation With Ryr Agonistsmentioning

confidence: 99%

“…Previous studies have demonstrated that EBV (Epstein-Barr virus)-immortalized human B-lymphocytes express the skeletal muscle isoform of the RyR [27,28] and that they could be used as a model to test the effect of mutations on RyR function. In order to shed light on the functional impact of RYR1 mutations linked to MmD, in the present study we investigated the influence of naturally occurring RYR1 mutations identified in patients with MmD and/or CCD, on intracellular Ca 2+ homoeostasis and pharmacological RyR activation, in EBV-immortalized B-lymphocytes.…”

Section: Introductionmentioning

confidence: 99%

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Functional properties of ryanodine receptors carrying three amino acid substitutions identified in patients affected by multi-minicore disease and central core disease, expressed in immortalized lymphocytes

Ducreux

Zorzato

Ferreiro

et al. 2006

Biochemical Journal

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More than 80 mutations in the skeletal muscle ryanodine receptor gene have been found to be associated with autosomal dominant forms of malignant hyperthermia and central core disease, and with recessive forms of multi-minicore disease. Studies on the functional effects of pathogenic dominant mutations have shown that they mostly affect intracellular Ca2+ homoeostasis, either by rendering the channel hypersensitive to activation (malignant hyperthermia) or by altering the amount of Ca2+ released subsequent to physiological or pharmacological activation (central core disease). In the present paper, we show, for the first time, data on the functional effect of two recently identified recessive ryanodine receptor 1 amino acid substitutions, P3527S and V4849I, as well as that of R999H, another substitution that was identified in two siblings that were affected by multi-minicore disease. We studied the intracellular Ca2+ homoeostasis of EBV (Epstein-Barr virus)-transformed lymphoblastoid cells from the affected patients, their healthy relatives and control individuals. Our results show that the P3527S substitution in the homozygous state affected the amount of Ca2+ released after pharmacological activation with 4-chloro-m-cresol and caffeine, but did not affect the size of the thapsigargin-sensitive Ca2+ stores. The other substitutions had no effect on either the size of the intracellular Ca2+ stores, or on the amount of Ca2+ released after ryanodine receptor activation; however, both the P3527S and V4849I substitutions had a small but significant effect on the resting Ca2+ concentration.

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Section: Mutation Screeningmentioning

confidence: 99%

Section: Intracellular Ca 2+ Measurementsmentioning

confidence: 99%

Section: Sensitivity Of Lymphoblastoid Cell Lines To Pharmacological Activation With Ryr Agonistsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Functional properties of ryanodine receptors carrying three amino acid substitutions identified in patients affected by multi-minicore disease and central core disease, expressed in immortalized lymphocytes

Ducreux

Zorzato

Ferreiro

et al. 2006

Biochemical Journal

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“…Prolonged ER stress can contribute to muscle atrophy, inflammation, insulin resistance, and disrupted proteostasis [43]. Increased release of pro-inflammatory cytokines IL-6 and IL-1 from cultured myotubes and B-lymphocytes has been observed in MH-causing RYR1 mutations [44,45], suggesting a potential interplay with the immune system. There is a fine balance between local, transient inflammation that stimulates pro-myogenic muscle repair and wide-spread, chronic inflammation that diminishes regenerative capability [46].…”

Section: Discussionmentioning

confidence: 99%

Identification of ER/SR resident proteins as biomarkers for ER/SR calcium depletion in skeletal muscle cells

Greer

Meilleur

Harvey

et al. 2021

Preprint

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Aberrations to endoplasmic/sarcoplasmic reticulum (ER/SR) calcium concentration can result in the departure of endogenous proteins in a phenomenon termed exodosis. Redistribution of the ER/SR proteome can have deleterious effects to cell function and cell viability, often contributing to disease pathogenesis. Many proteins prone to exodosis reside in the ER/SR via an ER retention/retrieval sequence (ERS) and are involved in protein folding, protein modification, and protein trafficking. While the consequences of their extracellular presence have yet to be fully delineated, the proteins that have undergone exodosis may be useful for biomarker development. Skeletal muscle cells rely upon tightly coordinated ER/SR calcium release for muscle contractions, and perturbations to calcium homeostasis can result in myopathies. Ryanodine receptor type-1 (RYR1) is a calcium release channel located in the SR. Mutations to the RYR1 gene can compromise calcium homeostasis leading to a vast range of clinical phenotypes encompassing hypotonia, myalgia, respiratory insufficiency, ophthalmoplegia, fatigue and malignant hyperthermia (MH). There are currently no FDA approved treatments for RYR1-related myopathies (RYR1-RM). Here we examine the exodosis profile of skeletal muscle cells following ER/SR calcium depletion. Proteomic analysis identified 4,465 extracellular proteins following ER/SR calcium depletion with 1280 proteins significantly different than vehicle. A total of 54 ERS proteins were identified and 33 ERS proteins significantly increased following ER/SR calcium depletion. Specifically, ERS protein, mesencephalic astrocyte-derived neurotrophic factor (MANF), was elevated following calcium depletion, making it a potential biomarker candidate for human samples. Despite no significant elevation of MANF in plasma levels among healthy volunteers and RYR1-RM individuals, MANF plasma levels positively correlated with age in RYR1-RM individuals, presenting a potential biomarker of disease progression. Selenoprotein N (SEPN1) was also detected only in extracellular samples following ER/SR calcium depletion. This protein is integral to calcium handling and SEPN1 variants have a causal role in SEPN1-related myopathies (SEPN1-RM). Extracellular presence of ER/SR membrane proteins may provide new insight into proteomic alterations extending beyond ERS proteins. Pre-treatment of skeletal muscle cells with bromocriptine, an FDA approved drug recently found to have anti-exodosis effects, curbed exodosis of ER/SR resident proteins. Changes to the extracellular content caused by intracellular calcium dysregulation presents an opportunity for biomarker development and drug discovery.

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Ryanodine receptor and immune‐related molecules in diabetic cardiomyopathy

et al. 2021

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Hyperglycaemia is a major aetiological factor in the development of diabetic cardiomyopathy. Excessive hyperglycaemia increases the levels of reactive carbonyl species (RCS), reactive oxygen species (ROS) and reactive nitrogen species (RNS) in the heart and causes derangements in calcium homeostasis, inflammation and immune-system disorders. Ryanodine receptor 2 (RyR2) plays a key role in excitation-contraction coupling during heart contractions, including rhythmic contraction and relaxation of the heart. Cardiac inflammation has been indicated in part though interleukin 1 (IL-1) signals, supporting a role for B and T lymphocytes in diabetic cardiomyopathy. Some of the post-translational modifications of the ryanodine receptor (RyR) by RCS, ROS and RNS stress are known to affect its gating and Ca 2+ sensitivity, which contributes to RyR dysregulation in diabetic cardiomyopathy. RyRs and immune-related molecules are important signalling species in many physiological and pathophysiological processes in various heart and cardiovascular diseases. However, little is known regarding the mechanistic relationship between RyRs and immune-related molecules in diabetes, as well as the mechanisms mediating complex communication among cardiomyocytes, fibroblasts and immune cells. This review highlights new findings on the complex cellular communications in the pathogenesis and progression of diabetic cardiomyopathy. We discuss potential therapeutic applications targeting RyRs and immune-related molecules in diabetic complications.

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B-lymphocytes from malignant hyperthermia-susceptible patients have an increased sensitivity to skeletal muscle by ryanodine receptor activators.

Cited by 9 publications

References 0 publications

Functional properties of ryanodine receptors carrying three amino acid substitutions identified in patients affected by multi-minicore disease and central core disease, expressed in immortalized lymphocytes

Functional properties of ryanodine receptors carrying three amino acid substitutions identified in patients affected by multi-minicore disease and central core disease, expressed in immortalized lymphocytes

Identification of ER/SR resident proteins as biomarkers for ER/SR calcium depletion in skeletal muscle cells

Ryanodine receptor and immune‐related molecules in diabetic cardiomyopathy

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