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Preclinical model systems of ryanodine receptor 1-related myopathies and malignant hyperthermia: a comprehensive scoping review of works published 1990–2019



Pathogenic variations in the gene encoding the skeletal muscle ryanodine receptor (RyR1) are associated with malignant hyperthermia (MH) susceptibility, a life-threatening hypermetabolic condition and RYR1-related myopathies (RYR1-RM), a spectrum of rare neuromuscular disorders. In RYR1-RM, intracellular calcium dysregulation, post-translational modifications, and decreased protein expression lead to a heterogenous clinical presentation including proximal muscle weakness, contractures, scoliosis, respiratory insufficiency, and ophthalmoplegia. Preclinical model systems of RYR1-RM and MH have been developed to better understand underlying pathomechanisms and test potential therapeutics.


We conducted a comprehensive scoping review of scientific literature pertaining to RYR1-RM and MH preclinical model systems in accordance with the PRISMA Scoping Reviews Checklist and the framework proposed by Arksey and O’Malley. Two major electronic databases (PubMed and EMBASE) were searched without language restriction for articles and abstracts published between January 1, 1990 and July 3, 2019.


Our search yielded 5049 publications from which 262 were included in this review. A majority of variants tested in RYR1 preclinical models were localized to established MH/central core disease (MH/CCD) hot spots. A total of 250 unique RYR1 variations were reported in human/rodent/porcine models with 95% being missense substitutions. The most frequently reported RYR1 variant was R614C/R615C (human/porcine total n = 39), followed by Y523S/Y524S (rabbit/mouse total n = 30), I4898T/I4897T/I4895T (human/rabbit/mouse total n = 20), and R163C/R165C (human/mouse total n = 18). The dyspedic mouse was utilized by 47% of publications in the rodent category and its RyR1-null (1B5) myotubes were transfected in 23% of publications in the cellular model category. In studies of transfected HEK-293 cells, 57% of RYR1 variations affected the RyR1 channel and activation core domain. A total of 15 RYR1 mutant mouse strains were identified of which ten were heterozygous, three were compound heterozygous, and a further two were knockout. Porcine, avian, zebrafish, C. elegans, canine, equine, and drosophila model systems were also reported.


Over the past 30 years, there were 262 publications on MH and RYR1-RM preclinical model systems featuring more than 200 unique RYR1 variations tested in a broad range of species. Findings from these studies have set the foundation for therapeutic development for MH and RYR1-RM.


Ryanodine receptor 1-related myopathies (RYR1-RM) are a diverse spectrum of rare monogenic neuromuscular disorders that manifest from variations in the RYR1 gene [1, 2]. In total, > 700 RYR1 variations have been identified; many of which are private to an individual case or family [3]. RYR1 exhibits little functional variation (per a recently developed bioinformatic residual variance intolerance [RVIS] scoring system: − 8.29 [0.01%]) [4] and encodes a 2.2 megadalton homotetrameric calcium ion channel (RyR1) that is localized to the sarcoplasmic reticulum (SR) membrane in skeletal muscle [5]. The physical connection between the RyR1 cytosolic shell and dihydropyridine receptor (DHPR) enables a coordinated release of SR calcium to the muscle cell cytosol, a process that facilitates excitation-contraction coupling in response to depolarization of the transverse-tubule membrane [6, 7]. ER/SR calcium concentration is an estimated 1000–10,000 times greater than cytosolic calcium concentration, and maintenance of this steep gradient is imperative to the health of the cell [8, 9]. Preclinical studies have identified intracellular calcium dysregulation as the central pathomechanism resulting from RYR1 variations characterized by SR calcium leak or excitation-contraction uncoupling [10]. In addition, the presence of truncation variations often reported in compound heterozygous cases can lead to decreased RyR1 expression [11, 12]. Owing to > 100 cysteine residues per subunit, RyR1 are susceptible to post-translational modifications, which in the case of mutant channels, further exacerbate intracellular calcium dysregulation though a previously reported feed-forward mechanism [13, 14]. For example, an elevated level of S-nitrosylated cysteines greatly increases channel activity, thus perpetuating calcium release. RYR1-RM pathomechanisms have been reviewed in detail elsewhere [10].

RYR1-RM can be inherited in a dominant or recessive manner and are slowly progressive with clinical manifestations including proximal muscle and facial weakness, joint contractures, scoliosis, ophthalmoplegia, and respiratory muscle weakness [15]. Although presentation often occurs at birth or in early childhood, adult-onset cases have also been reported [16, 17]. Affected individuals are considered at risk of malignant hyperthermia (MH) susceptibility. Genetic predisposition to MH can result in a potentially fatal hypermetabolic response and skeletal muscle rigidity upon exposure to triggers such as volatile anesthetics, exercise in the heat, and influenza [18, 19]. In addition to myopathy, other clinical phenotypes attributed to RYR1 variations include rhabdomyolysis-myalgia syndrome and intermittent periodic paralysis [20, 21]. Historically, RYR1-RM were sub-categorized based on skeletal muscle histopathology. This yielded subtypes such as central core disease, multiminicore disease, centronuclear myopathy, and congenital fiber-type disproportion [22]. Despite being the most frequently reported non-dystrophic neuromuscular disorder [23], there is currently no approved treatment for RYR1-RM.

A decade after the first report of central core disease in humans [24], Hall and colleagues observed a fatal hypermetabolic response to suxamethonium in pigs [25]. This was the first MH animal model system whose phenotype, also referred to as porcine stress syndrome, was later attributed to the R615C variation in RYR1 [26, 27]. Since this landmark discovery, technological and scientific advances have led to the development of preclinical model systems that can be grouped into cell culture and animal categories, each with their own advantages and limitations [28,29,30,31,32].


The objective of this scoping review was to comprehensively review the scientific literature for MH and RYR1-RM preclinical model systems, thus generating a resource to guide future research.


The PRISMA extension for Scoping Reviews (PRISMA-ScR) Checklist and the framework proposed by Arksey and O’Malley [33] were used to guide this scoping review. The overarching research question was: what preclinical model systems have been reported for MH and RYR1-RM?

Identifying relevant studies

Two major electronic databases (PubMed and EMBASE) were searched without language restriction for articles and abstracts published between January 1, 1990 and July 3, 2019. The search strategy comprised the following a priori search terms present in the title or abstract using Boolean operators and MeSH terms: RYR-1 OR RYR1 OR RyR1s OR ryanodine receptor calcium release channel OR “ryanodine receptor 1” AND malignant hyperthermia OR “malignant hyperthermia” OR malignant hyperpyrexia OR anesthesia hyperthermia OR Muscular diseases OR muscular diseases OR myopathies OR myopathy OR muscle OR muscular OR muscle contraction OR muscle contraction OR smooth muscle OR cardiac muscle OR skeletal muscle OR muscle fiber OR myofibril. The full search strategy is provided in Additional file 1.

Study selection

Following removal of duplicates, titles and abstracts of all publications were reviewed independently by two of the authors and marked for inclusion if they discussed a MH or RYR1-RM preclinical model system. Publications were marked for exclusion if they were (1) not gene or isoform of interest (e.g. CACNA1S-related MH), (2) clinical report, (3) structural biology, (4) wild-type models and methods, (5) cardiac or smooth muscle, (6) review articles, or (7) categorized as miscellaneous. All publications were discussed with a third author who adjudicated when there was discordance between the first two authors over whether publication should be included or excluded.

Charting data and reporting the results

The following data were extracted from full text publications selected for inclusion in the review: first author, year of publication, title of the publication, variation(s) of the preclinical model system(s), and conclusions of the publication on the disease model system(s). Data were tabulated according to type of preclinical model system. Categories included transfected human embryonic kidney cell (HEK)-293 cells, transfected RYR1-null (dyspedic) myotubes, immortalized B-lymphocytes, primary cell culture, porcine model systems, and rodent model systems. Data on all other preclinical model systems, including zebrafish, avian, C. elegans, and drosophila, were combined and tabulated separately. Two authors reviewed data extracted for each article. To identify gaps in the literature where no preclinical model system had been reported for a specific RYR1 protein-coding region, the number of publications per RYR1 exon was mapped against established MH/CCD hotspot regions and sequence of the RyR1 protein structure. The composition of included and excluded publications was also summarized.


Study characteristics

The search strategy utilized in this study yielded 5049 research publications between January 1, 1990 and July 3, 2019. Nine additional publications were retrieved through other information sources. Following removal of 2814 duplicates, 2284 abstracts were screened for inclusion. A total of 1956 publications were excluded at this point, leaving 328 for full text review. During full text review, 66 additional publications were excluded leaving 262 publications for inclusion in this review. An overview of this process is provided in Fig. 1.

Fig. 1

PRISMA diagram summarizing the article selection workflow

The majority of publications that met inclusion criteria for this review focused on RYR1 cellular and rodent model systems (43 and 39%, and respectively), Fig. 2a. Wild-type/methods publications formed the largest group of those excluded (24%), followed by those focused on cardiac/smooth muscle (19%), not isoform/gene of interest (16%), and clinical reports (13%), Fig. 2b.

Fig. 2

a-b Composition of included and excluded publications

The highest frequencies of variations reported in RYR1 preclinical model systems were localized to established MH/central core disease (MH/CCD) hot spots 1, 2, and 3 located between exons 1–17, 39–46, and 90–103, respectively, Fig. 3. At least one RYR1 preclinical model system was reported for every RyR1 structural region.

Fig. 3

Number of publications per RYR1 exon aligned with corresponding MH/CCD hotspots and RyR1 structural regions

A total of 250 unique RYR1 variations were reported in human/mouse/porcine model systems with 95% being missense substitutions. The most frequently reported RYR1 variations reported across species were R614C/R615C (human/porcine total n = 39), Y523S/Y524S (rabbit/mouse total n = 30), I4898T/I4897T/I4895T (human/rabbit/mouse total n = 20), and R163C/R165C (human/mouse total n = 18). The dyspedic mouse was the most frequently reported mouse model system comprising 47% of publications in this category. The predominant type of RYR1 preclinical model system used has varied over time. From 1990 to 1994, the R615C porcine model system was most frequently reported. Cellular model systems were then most frequently reported until 2010, after which this transitioned to rodent model systems including RyR1-null (dyspedic) and Y524S, R163C, and I4895T mutant mice, Fig. 4.

Fig. 4

Total number of publications over time and type of RYR1 preclinical model system reported

Cellular model systems

Expression of recombinant RYR1 in heterologous cells

A total of 49 publications reported transfecting mutant RYR1 cDNA into HEK-293 cells, which lack native RyR1 channels, making this the most frequently utilized cellular model, Table 1. These 49 publications reported on 161 unique RYR1 variations of which 153 were missense substitutions, six were deletions, one was a frameshift variant resulting in a truncation, and one was a deletion-insertion resulting in a truncation. Of these unique variations, 57% affected the RyR1 channel and activation core domain. In the 49 publications reporting on mutant HEK-293 cells, 13 variations were evaluated and/or functionally characterized (at least three times) (C36R [47, 81, 83], R164C [43, 47, 81, 83], G249R [47, 81, 83], G342R [39, 47, 81, 83], Y523S [43, 47, 70, 81, 83], R615C [47, 70, 81, 83], R2163C [45, 81, 83], R2163H [45, 81, 83], R2435H [39, 43, 45], R2458C [45, 81, 83], R2458H [45, 81, 83], R2508C [45, 52, 61], R2508H [45, 48, 52]). A majority (13/14) of these well-characterized missense substitutions affected the RyR1 cytosolic shell domain. In four publications, multiple variants were introduced to HEK-293 cells to evaluate their impact alone and in combination on RyR1 structural conformation and calcium homeostasis [35,36,37, 67]. Two additional publications reported on monkey-derived CV-1 in Origin with SV40 (COS)-7 cells that were transfected with mutant RYR1 cDNA [84, 85].

Table 1 Cellular RYR1 model systems: Human embryonic kidney (HEK-293) cells

Expression of recombinant RYR1 in dyspedic myotubes

Transfection of dyspedic myotubes with mutant RYR1 cDNA was reported in 25 publications, Table 2, in which a total of 49 unique variations were tested. This includes studies that used the 1B5 cell line, derived by transduction of dyspedic mouse fibroblasts with MyoD, to evaluate mutant RyR1 channel function [109]. Of these 49 variations, 44 were missense substitutions and 5 were deletions with a majority (27/49) affecting the RyR1 channel and activation core domain. One missense substitution, E4032A [103, 105, 106], was evaluated and/or functionally characterized at least three times in transfected dyspedic myotubes.

Table 2 Cellular RYR1 model systems: Transfected RYR1-null (dyspedic) myotubes

Expression of endogenous mutant RYR1

Additionally, 16 publications reported immortalization of patient primary B-lymphocytes for downstream functional characterization, Table 3. These 16 publications included 32 unique missense substitutions, one deletion, and two deletion-insertions. A total of 50 unique RYR1 variants, all missense substitutions, were tested in 19 publications utilizing primary cell culture model systems, Table 4.

Table 3 Cellular RYR1 model systems: Immortalized B-lymphocytes
Table 4 Primary cell culture model systems

Animal model systems


A total of 15 RYR1 rodent model systems were identified of which ten were heterozygous, three were compound heterozygous, and a further two were knockout, Table 5. Variations discussed in this section are numbered according to the mouse sequence. Core formation was reported in three of the rodent model systems, excluding knockout (Y524S [158], Q1970fsX16 + A4329D [179], I4895T [168]). Overall, six of the ten heterozygous rodent model systems had missense substitutions affecting the RyR1 cytosolic shell domain. Two compound heterozygous model systems had a single missense substitution engineered into one allele with a frameshift leading to a deletion or truncation on the opposite allele [178, 179]. In these model systems, one variation affected the RyR1 cytosolic shell and the other affected the RyR1 channel and activation core. An additional compound heterozygous model system had a single missense substitution affecting the RyR1 channel and activation core with a second missense substitution and deletion, on the opposite allele, affecting the RyR1 cytosolic shell [181]. Various forms of aberrant intracellular calcium dynamics were reported in all rodent systems (except knockout). This included evidence of increased resting cytosolic calcium and RyR1-open probability under resting conditions (SR calcium leak) [173] as well as decreased calcium permeation (excitation-contraction uncoupling) [30]. The two most frequently reported RYR1 rodent model systems were the dyspedic mouse, accounting for 47% of rodent publications [54, 109, 194,195,196,197,198,199,200,201,202,203,204,205,206,207,208,209,210,211,212,213,214,215,216,217,218,219,220,221,222,223,224,225,226,227,228,229,230,231,232,233,234,235,236,237,238], and the Y524S knock-in mouse, which accounted for 22% of rodent publications, Table 5. Studies utilizing dyspedic mice/1B5 myotubes not transfected with mutant RYR1 cDNA, were primarily focused on elucidating the following: (a) relative importance and functional role of wild-type RyR isoforms [213, 227, 234], (b) fundamental physiology of excitation-contraction coupling components [205, 225, 232], (c) roles of specific RyR1 structural regions on channel function [216, 222, 235]. The Y524S knock-in mouse has been utilized extensively to investigate the mechanisms behind several phenotypes on the RYR1-RM disease spectrum including MH susceptibility [162], statin-induced myopathy [152], and central core disease [158]. Y524S mice have also been used to test potential therapeutics for RYR1-RM including the antioxidant N-acetylcysteine [145, 160] and the activator of the AMP-activated protein kinase 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) [153].

Table 5 Rodent RYR1 model systems

Other animal model systems

The pathomechanism, diagnosis, and acute treatment of malignant hyperthermia was investigated in 24 publications that used the R615C porcine model system [27, 239,240,241,242,243,244,245,246,247,248,249,250,251,252,253,254,255,256,257,258,259,260,261], Table 6. A number of other preclinical model systems have been described including avian, zebrafish, C. elegans, canine, equine, and drosophila, Table 7. Six publications reported on a single recessive zebrafish model system of RYR1-RM termed the relatively relaxed (ryrmi340) mutant [28, 263,264,265,266,267] which was utilized for high-throughput drug screening [263] and testing of N-acetylcysteine as a potential therapeutic to address elevated oxidative stress [264]. A further six publications reported using Caenorhabditis elegans (C. elegans) with variants in unc68, the RYR1 ortholog [32, 280,281,282,283,284]. With 40% sequence homology to humans, C. elegans have been used to investigate RyR1 functional sites [281] and test the potential impact of RYR1 mutations on central nervous system function [32]. A single heterozygous canine model system of malignant hyperthermia was reported. The canine model system carried a single missense substitution, V547A, affecting the RyR1 cytosolic shell domain and was characterized by responsiveness to an in vivo halothane-succinylcholine challenge and having a positive in vitro contracture test [278]. Four publications described equine model systems of malignant hyperthermia and exertional rhabdomyolysis that carried variations in the RYR1 gene [274,275,276,277]. Two RYR1 variants were reported: (a) R2454G associated with fulfilment malignant hyperthermia and a high affinity for ryanodine binding [277] and (b) C7360G associated with both anesthetic-induced malignant hyperthermia and exertional/non-exertional rhabdomyolysis [276]. Three publications reported on drosophila with variations in the equivalent RYR1 gene (dRyr) [271,272,273]. A total of nine RYR1 variations were presented comprising eight missense substitutions and one insertion, Table 7. Missense substitutions in drosophila dRyr conferred halothane sensitivity [272], and drosophila with CRISPR/Cas9 gene-edited dRyr have been used to investigate insecticide resistance [271]. Three publications utilized transfected wild-type rodent cells to generate RYR1 model systems with clinically-relevant variations [268,269,270] and four reported on the avian crooked neck dwarf mutant which lacks the alpha RyR isoform homologous to human RyR1 [285,286,287,288].

Table 6 Porcine RYR1 model system of malignant hyperthermia
Table 7 Other RYR1 preclinical model systems


This comprehensive scoping review of MH and RYR1-RM preclinical model systems identified 262 relevant published records and serves as a compendium to guide future research. During the period spanning January 1, 1990 to July 3, 2019 a diverse range of preclinical model systems were utilized to investigate the etiology, pathomechanisms, and potential treatments for MH and RYR1-RM. There has been sustained research output since 2010 with the predominant model system used varying over time between porcine, cellular, and rodent.

A single missense substitution, R615C, was the sole porcine variant reported. As the first RYR1 preclinical model system, studies of R615C pigs led to fundamental discoveries including identification of 4-CmC as a potent RyR1 agonist and identification of RYR1 as a genetic locus for malignant hyperthermia [27, 255]. The R615C porcine model system was also utilized to better understand the mechanism of dantrolene which remains the only approved treatment for MH crises [252].

The number of RYR1 variations reported in the literature (> 700) has been prohibitive in terms of developing in vivo model systems reflecting each variant. This review has outlined the extent to which cellular model systems, in particular transfected HEK-293 cells and dyspedic myotubes, have been versatile systems through which to investigate the pathogenicity of RYR1 variations and their impact on intracellular calcium homeostasis. However activity of the RyR1 protein complex is tightly regulated by coupling to the dihydropyridine receptor and by modulators of channel function such as 12-kDa FK506-binding protein (FKBP12) and calmodulin [10]. Absence of these components in the HEK-293 system may therefore affect the reliability of functional data for clinical translation. Epstein-Barr virus-driven immortalization of patient-derived lymphoblasts has also proven a valuable non-recombinant methodology when clinical biospecimens are available, although they also do not contain all elements of the skeletal muscle triad. Both HEK-293 cells and dyspedic myotubes have a standardized and well-characterized background and are therefore less likely, than immortalized patient cells, to be influenced by variations in other genes that may impact RyR1 function. Although patient tissue is not always readily available, it is important to recognize that functional studies of patient-derived primary myotubes can provide valuable supporting evidence of RYR1 variant pathogenicity. Indeed, such studies have been incorporated within the MH variant scoring matrix developed by the European Malignant Hyperthermia Group (EMHG) [289]. Despite the above limitations, recent advances in the engineering of skeletal muscle three-dimensional systems using patient-derived induced pluripotent stem cells holds the prospect of providing a physiologically relevant cellular system through which to evaluate and screen potential treatments for skeletal muscle disorders, including RYR1-RM [290,291,292]. Our observation that the most frequently reported variants were localized to MH/CCD hotspot regions is consistent with the initial clinical focus to identify patients with variants in these distinct regions and perform functional characterization [293]. Common functional analyses identified in this review include RyR1 agonist sensitivity (caffeine, 4-CmC), 3[H]-ryanodine binding, halothane and/or isoflurane sensitivity, and intracellular calcium measurements via calcium-sensitive fluorescent dyes such as fluo-4.

The dyspedic mouse was utilized by 47% of publications in the rodent category and its RYR1-null myotubes were transfected in 23% of publications in the cellular model category, a testament to importance of the dyspedic mouse for both understanding the fundamental physiology of the ryanodine receptor and as a stable model system to characterize mutant RyR1 channels. Heterozygous knock-in rodent models have formed the basis of in vivo RYR1-RM preclinical testing. Y524S, I4895T, and R163C were the most extensively studied knock-in rodent models over the last 30 years. These mice have provided valuable insights into the effects of single missense substitutions on RyR1 dysfunction including channel leak and excitation-contraction uncoupling. Furthermore, these mice have enabled the identification downstream pathologic sequalae in vivo such as elevated oxidative/nitrosative/ER stress and an unfolded protein response. However the abovementioned knock-in mice do not necessarily mirror the phenotypes observed in autosomal dominant patients with equivalent RYR1 variants (reviewed in detail elsewhere [292]). Two recently published compound heterozygous RYR1-RM rodent models recapitulate clinical manifestations observed in recessive RYR1-RM patients, including decreased RyR1 protein expression, reduced muscle mass, and progressive muscle weakness [178, 179]. An additional compound heterozygous mouse (T4706M/S1669C + L1716del) included in this review is currently undergoing full characterization [181].

In contrast to rodent model systems, zebrafish are more cost-efficient, have transparent embryos that facilitate visualization of dynamic events, have a shorter lifecycle, have larger hatch sizes, and are easier to maintain [294, 295]. Zebrafish are readily manipulated by chemical approaches because embryos can readily absorb compounds that they are exposed to in solution, therefore allowing for high-throughput chemical screening [296, 297]. A recessive zebrafish model system of RYR1-RM, termed the relatively relaxed (ryrmi340) mutant, exhibits weak muscle contractions resulting in slow swimming, dramatically decreased Ca2+ transients at the t-tubules of fast muscles due to defective E-C coupling, and small amorphous cores detectable by electron microscopy. Despite the abovementioned advantages over rodent model systems, a consideration is that relatively relaxed (ryr1b) zebrafish are homozygous with a truncated RyR1 channel (residual expression = 1–10% of normal RyR1). As such, its genetic defect and pathomechanism do not align with a majority of RYR1-RM clinical cases.

Consistent with the findings of this review, over 90% of animals used in research are mice or rats [298]. However, other animal model systems have also been developed and used to study the skeletal muscle ryanodine receptor and the consequences of genetic variations. C. elegans and drosophila have been primarily used for genetic and developmental biology studies, whereas the porcine, equine, and canine model systems have focused on understanding and characterizing the etiology of MH in these species. As an alternative to higher order RYR1-RM animal model systems, the use of simpler organisms (C. elegans, yeast, drosophila) and vertebrates (in particular zebrafish) with sufficient genome sequence homology to humans could be revitalized using more precise genome editing techniques such as prime editing [299]. However, due to evolutionary distance between DNA sequences, results from non-mammalian model systems should undergo further careful validation in mammals such as mice and pigs prior to translation to clinical studies. It is possible that records published in supplementary material may not have been captured by the search strategy used for this review and may be considered a limitation.

Advances in functional genomics coupled with the increase in demand for mice as a primary experimental system are expected to continue driving the need for additional transgenic, gene-edited and combinatorial breeding of different RYR1-RM model systems in the near future. Development of conditional targeted animal models (cre/lox, tet, and other similar approaches) can reduce generation and retention of extraneous animals and also allow for the conduct of developmental studies in late-onset myopathy subtypes in this heterogeneous group of disorders. Determining which murine model most closely represents a majority of either dominant or recessive cases of RYR1-RM remains an open question. Funding of research utilizing recently developed model systems is essential to translating these promising advances into clinical trials and treatment discoveries.


Over the past 30 years, there were 262 publications on MH and RYR1-RM preclinical model systems featuring more than 200 unique RYR1 variations tested in a broad range of species. Findings from these studies have set the foundation for therapeutic development for MH and RYR1-RM.

Availability of data and materials

Not applicable.





5-aminoimidazole-4-carboxamide ribonucleoside




Allele-specific gene silencing

C. elegans :

Caenorhabditis elegans


Calcium-bound calmodulin




Central Core Disease


Calcium-induced calcium release


Congenital neuromuscular disease with uniform type 1 fiber


CV-1 in Origin with SV40 cell line


Dihydropyridine receptor


Epstein-Barr virus




Excitation-coupled calcium entry


European Malignant Hyperthermia Group


Flexor digitorum brevis


12-kDa FK506-binding protein


Human embryonic kidney cell line




Malignant hyperthermia


Multiminicore disease


Nitrous oxide


Preferred Reporting Items for Systematic Reviews and Meta-Analyses


Reactive oxygen species


Residual variance intolerance score


RYR1-related myopathies


Store overload-induced calcium release


Sarcoplasmic reticulum


Unfolded protein response


Voltage-gated calcium release


Wild type


  1. 1.

    Kaplan J-C, Hamroun D, Rivier F, Bonne G. The 2017 version of the gene table of monogenic neuromuscular disorders (nuclear genome). Neuromuscul Disord. 2016;26(12):895–929.

  2. 2.

    Lawal TA, Todd JJ, Meilleur KG. Ryanodine receptor 1-related myopathies: diagnostic and therapeutic approaches. Neurotherapeutics. 2018;15(4):885–99.

  3. 3.

    Litman RS, Griggs SM, Dowling JJ, Riazi S. Malignant hyperthermia susceptibility and related diseases. Anesthesiology. 2018;128(1):159–67.

  4. 4.

    Petrovski S, Wang Q, Heinzen EL, Allen AS, Goldstein DB. Genic intolerance to functional variation and the interpretation of personal genomes. PLoS Genet. 2013;9(8):e1003709.

  5. 5.

    des Georges A, Clarke OB, Zalk R, Yuan Q, Condon KJ, Grassucci RA, Hendrickson WA, Marks AR, Frank J. Structural basis for gating and activation of RyR1. Cell. 2016;167(1):145–157.e117.

  6. 6.

    Rebbeck RT, Karunasekara Y, Gallant EM, Board PG, Beard NA, Casarotto MG, Dulhunty AF. The β(1a) subunit of the skeletal DHPR binds to skeletal RyR1 and activates the channel via its 35-residue C-terminal tail. Biophys J. 2011;100(4):922–30.

  7. 7.

    Hernández-Ochoa E, Pratt S, Lovering R, Schneider M. Critical role of intracellular RyR1 calcium release channels in skeletal muscle function and disease. Front Physiol. 2016;6:420.

  8. 8.

    Henderson MJ, Wires ES, Trychta KA, Richie CT, Harvey BK. SERCaMP: a carboxy-terminal protein modification that enables monitoring of ER calcium homeostasis. Mol Biol Cell. 2014;25(18):2828–39.

  9. 9.

    Samtleben S, Jaepel J, Fecher C, Andreska T, Rehberg M, Blum R. Direct imaging of ER calcium with targeted-esterase induced dye loading (TED). J Vis Exp. 2013;75:e50317.

  10. 10.

    Witherspoon JW, Meilleur KG. Review of RyR1 pathway and associated pathomechanisms. Acta Neuropathol Commun. 2016;4(1):121.

  11. 11.

    Zhou H, Jungbluth H, Sewry CA, Feng L, Bertini E, Bushby K, Straub V, Roper H, Rose MR, Brockington M, et al. Molecular mechanisms and phenotypic variation in RYR1-related congenital myopathies. Brain. 2007;130(Pt 8):2024–36.

  12. 12.

    Monnier N, Marty I, Faure J, Castiglioni C, Desnuelle C, Sacconi S, Estournet B, Ferreiro A, Romero N, Laquerriere A, et al. Null mutations causing depletion of the type 1 ryanodine receptor (RYR1) are commonly associated with recessive structural congenital myopathies with cores. Hum Mutat. 2008;29(5):670–8.

  13. 13.

    Meissner G. The structural basis of ryanodine receptor ion channel function. J Gen Physiol. 2017;149(12):1065–89.

  14. 14.

    Michelucci A, De Marco A, Guarnier FA, Protasi F, Boncompagni S. Antioxidant treatment reduces formation of structural cores and improves muscle function in RYR1Y522S/WT mice. Oxidative Med Cell Longev. 2017;2017:15.

  15. 15.

    North KN, Wang CH, Clarke N, Jungbluth H, Vainzof M, Dowling JJ, Amburgey K, Quijano-Roy S, Beggs AH, Sewry C, et al. Approach to the diagnosis of congenital myopathies. Neuromuscul Disord. 2014;24(2):97–116.

  16. 16.

    Loseth S, Voermans NC, Torbergsen T, Lillis S, Jonsrud C, Lindal S, Kamsteeg EJ, Lammens M, Broman M, Dekomien G, et al. A novel late-onset axial myopathy associated with mutations in the skeletal muscle ryanodine receptor (RYR1) gene. J Neurol. 2013;260(6):1504–10.

  17. 17.

    Jungbluth H, Lillis S, Zhou H, Abbs S, Sewry C, Swash M, Muntoni F. Late-onset axial myopathy with cores due to a novel heterozygous dominant mutation in the skeletal muscle ryanodine receptor (RYR1) gene. Neuromuscul Disord. 2009;19(5):344–7.

  18. 18.

    Denborough M. Malignant hyperthermia. Lancet. 1998;352(9134):1131–6.

  19. 19.

    Zvaritch E, Gillies R, Kraeva N, Richer M, Jungbluth H, Riazi S. Fatal awake malignant hyperthermia episodes in a family with malignant hyperthermia susceptibility: a case series. Can J Anaesth. 2019;66:540.

  20. 20.

    Witting N, Laforet P, Voermans NC, Roux-Buisson N, Bompaire F, Rendu J, Duno M, Feillet F, Kamsteeg EJ, Poulsen NS, et al. Phenotype and genotype of muscle ryanodine receptor rhabdomyolysis-myalgia syndrome. Acta Neurol Scand. 2018;137(5):452–61.

  21. 21.

    Matthews E, Neuwirth C, Jaffer F, Scalco RS, Fialho D, Parton M, Raja Rayan D, Suetterlin K, Sud R, Spiegel R, et al. Atypical periodic paralysis and myalgia: a novel RYR1 phenotype. Neurology. 2018;90(5):e412–8.

  22. 22.

    Snoeck M, van Engelen BGM, Küsters B, Lammens M, Meijer R, Molenaar JPF, Raaphorst J, Verschuuren-Bemelmans CC, Straathof CSM, Sie LTL, et al. RYR1-related myopathies: a wide spectrum of phenotypes throughout life. Eur J Neurol. 2015;22(7):1094–112.

  23. 23.

    Amburgey K, McNamara N, Bennett LR, McCormick ME, Acsadi G, Dowling JJ. Prevalence of congenital myopathies in a representative pediatric United States population. Ann Neurol. 2011;70(4):662–5.

  24. 24.

    Magee KR, Shy GM. A new congenital non-progressive myopathy. Brain. 1956;79(4):610–21.

  25. 25.

    Hall LW, Woolf N, Bradley JW, Jolly DW. Unusual reaction to suxamethonium chloride. Br Med J. 1966;2(5525):1305.

  26. 26.

    Nelson TE, Jones EW, Anderson IL. Porcine malignant hyperthermia. Am J Pathol. 1976;84(1):197–200.

  27. 27.

    Fujii J, Otsu K, Zorzato F, de Leon S, Khanna VK, Weiler JE, O'Brien PJ, MacLennan DH: Identification of a mutation in porcine ryanodine receptor associated with malignant hyperthermia. Science (New York, NY) 1991, 253(5018):448–451.

  28. 28.

    Dowling JJ, Arbogast S, Hur J, Nelson DD, McEvoy A, Waugh T, Marty I, Lunardi J, Brooks SV, Kuwada JY, et al. Oxidative stress and successful antioxidant treatment in models of RYR1-related myopathy. Brain. 2012;135(Pt 4):1115–27.

  29. 29.

    Chelu MG, Goonasekera SA, Durham WJ, Tang W, Lueck JD, Riehl J, Pessah IN, Zhang P, Bhattacharjee MB, Dirksen RT, et al. Heat- and anesthesia-induced malignant hyperthermia in an RyR1 knock-in mouse. FASEB J. 2005;20(2):329–30.

  30. 30.

    Zvaritch E, Depreux F, Kraeva N, Loy RE, Goonasekera SA, Boncompagni S, Kraev A, Gramolini AO, Dirksen RT, Franzini-Armstrong C, et al. An Ryr1I4895T mutation abolishes Ca2+ release channel function and delays development in homozygous offspring of a mutant mouse line. Proc Natl Acad Sci U S A. 2007;104(47):18537–42.

  31. 31.

    Ruiz A, Eckhardt J, Elbaz M, Treves S, Zorzato F, Pelczar P, Muntoni F, Boncompagni S. Quantitative reduction of RyR1 protein caused by a single-allele frameshift mutation in RYR1 ex36 impairs the strength of adult skeletal muscle fibres; 2019.

  32. 32.

    Nicoll BK, Ferreira C, Hopkins PM, Shaw M-A, Hope IA. Aging effects of caenorhabditis elegans ryanodine receptor variants corresponding to human myopathic mutations. G3. 2017;7(5):1451–61.

  33. 33.

    Arksey H, O'Malley L. Scoping studies: towards a methodological framework. Int J Soc Res Methodol. 2005;8(1):19–32.

  34. 34.

    Chirasani VR, Xu L, Addis HG, Pasek DA, Dokholyan NV, Meissner G, Yamaguchi N. A central core disease mutation in the Ca(2+)-binding site of skeletal muscle ryanodine receptor impairs single-channel regulation. Am J Phys Cell Phys. 2019;317(2):C358–c365.

  35. 35.

    Xu L, Mowrey DD, Chirasani VR, Wang Y, Pasek DA, Dokholyan NV, Meissner G. G4941K substitution in the pore-lining S6 helix of the skeletal muscle ryanodine receptor increases RyR1 sensitivity to cytosolic and luminal ca(2). J Biol Chem. 2018;293(6):2015–28.

  36. 36.

    Xu L, Chirasani VR, Carter JS, Pasek DA, Dokholyan NV, Yamaguchi N, Meissner G. Ca(2+)-mediated activation of the skeletal-muscle ryanodine receptor ion channel. J Biol Chem. 2018;293(50):19501–9.

  37. 37.

    Xu L, Wang Y, Yamaguchi N, Pasek DA, Meissner G. Single channel properties of heterotetrameric mutant RyR1 ion channels linked to core myopathies. J Biol Chem. 2008;283(10):6321–9.

  38. 38.

    Schiemann AH, Bjorksten AR, Gillies RL, Hockey BM, Ball C, Pollock N, Bulger T, Stowell KM. A genetic mystery in malignant hyperthermia ‘solved’? Br J Anaesth. 2018;121(3):681–2.

  39. 39.

    Murayama T, Kurebayashi N, Ishigami-Yuasa M, Mori S, Suzuki Y, Akima R, Ogawa H, Suzuki J, Kanemaru K, Oyamada H, et al. Efficient high-throughput screening by endoplasmic reticulum Ca(2+) measurement to identify inhibitors of ryanodine receptor Ca(2+)-release channels. Mol Pharmacol. 2018;94(1):722–30.

  40. 40.

    Kondo T, Yasuda T, Mukaida K, Otsuki S, Kanzaki R, Miyoshi H, Hamada H, Nishino I, Kawamoto M. Genetic and functional analysis of the RYR1 mutation p.Thr84Met revealed a susceptibility to malignant hyperthermia. J Anesth. 2018;32(2):174–81.

  41. 41.

    Parker R, Schiemann AH, Langton E, Bulger T, Pollock N, Bjorksten A, Gillies R, Hutchinson D, Roxburgh R, Stowell KM. Functional characterization of C-terminal ryanodine receptor 1 variants associated with central core disease or malignant hyperthermia. J Neuromuscul Dis. 2017;4(2):147–58.

  42. 42.

    Merritt A, Booms P, Shaw MA, Miller DM, Daly C, Bilmen JG, Stowell KM, Allen PD, Steele DS, Hopkins PM. Assessing the pathogenicity of RYR1 variants in malignant hyperthermia. Br J Anaesth. 2017;118(4):533–43.

  43. 43.

    Chen W, Koop A, Liu Y, Guo W, Wei J, Wang R, MacLennan DH, Dirksen RT, Chen SRW. Reduced threshold for store overload-induced Ca(2+) release is a common defect of RyR1 mutations associated with malignant hyperthermia and central core disease. Biochem J. 2017;474(16):2749–61.

  44. 44.

    Stephens J, Schiemann AH, Roesl C, Miller D, Massey S, Pollock N, Bulger T, Stowell K. Functional analysis of RYR1 variants linked to malignant hyperthermia. Temperature (Austin). 2016;3(2):328–39.

  45. 45.

    Murayama T, Kurebayashi N, Ogawa H, Yamazawa T, Oyamada H, Suzuki J, Kanemaru K, Oguchi K, Iino M, Sakurai T. Genotype-phenotype correlations of malignant hyperthermia and central core disease mutations in the central region of the RYR1 channel. Hum Mutat. 2016;37(11):1231–41.

  46. 46.

    Gomez AC, Holford TW, Yamaguchi N. Malignant hyperthermia-associated mutations in the S2-S3 cytoplasmic loop of type 1 ryanodine receptor calcium channel impair calcium-dependent inactivation. Am J Phys Cell Phys. 2016;311(5):C749–c757.

  47. 47.

    Murayama T, Kurebayashi N, Yamazawa T, Oyamada H, Suzuki J, Kanemaru K, Oguchi K, Iino M, Sakurai T. Divergent activity profiles of type 1 ryanodine receptor channels carrying malignant hyperthermia and central Core disease mutations in the amino-terminal region. PLoS One. 2015;10(6):e0130606.

  48. 48.

    Miyoshi H, Yasuda T, Otsuki S, Kondo T, Haraki T, Mukaida K, Nakamura R, Hamada H, Kawamoto M. Several ryanodine receptor type 1 gene mutations of p.Arg2508 are potential sources of malignant hyperthermia. Anesth Analg. 2015;121(4):994–1000.

  49. 49.

    Mei Y, Xu L, Mowrey DD, Mendez Giraldez R, Wang Y, Pasek DA, Dokholyan NV, Meissner G. Channel gating dependence on pore lining helix glycine residues in skeletal muscle ryanodine receptor. J Biol Chem. 2015;290(28):17535–45.

  50. 50.

    Shirvanyants D, Ramachandran S, Mei Y, Xu L, Meissner G, Dokholyan NV. Pore dynamics and conductance of RyR1 transmembrane domain. Biophys J. 2014;106(11):2375–84.

  51. 51.

    Roesl C, Sato K, Schiemann A, Pollock N, Stowell KM. Functional characterisation of the R2452W ryanodine receptor variant associated with malignant hyperthermia susceptibility. Cell Calcium. 2014;56(3):195–201.

  52. 52.

    Miyoshi H, Haraki T, Yasuda T, Mukaida K, Hamada H, Kawamoto M. Two different variants of p.2508 in Japanese malignant hyperthermia patients causing hypersensitivity of ryanodine receptor 1: 7AP6-5. Eur J Anaesthesiol. 2014;31:123.

  53. 53.

    Sato K, Roesl C, Pollock N, Stowell KM. Skeletal muscle ryanodine receptor mutations associated with malignant hyperthermia showed enhanced intensity and sensitivity to triggering drugs when expressed in human embryonic kidney cells. Anesthesiology. 2013;119(1):111–8.

  54. 54.

    Kraeva N, Zvaritch E, Rossi AE, Goonasekera SA, Zaid H, Frodis W, Kraev A, Dirksen RT, Maclennan DH, Riazi S. Novel excitation-contraction uncoupled RYR1 mutations in patients with central core disease. Neuromuscul Disord. 2013;23(2):120–32.

  55. 55.

    Merritt A, Booms P, Fisher N, Duke AM, Sato K, Kim J, Jarvik GP, Roiz De Sa D, Stowell K, Steele D, et al. Functional analysis of the p.D1056H RYR1 variant associated with malignant hyperthermia and exertional heat stroke. Br J Anaesth. 2012;109(4):663.

  56. 56.

    Murayama T, Kurebayashi N, Oba T, Oyamada H, Oguchi K, Sakurai T, Ogawa Y. Role of amino-terminal half of the S4-S5 linker in type 1 ryanodine receptor (RyR1) channel gating. J Biol Chem. 2011;286(41):35571–7.

  57. 57.

    Haraki T, Yasuda T, Mukaida K, Migita T, Hamada H, Kawamoto M. Mutated p.4894 RyR1 function related to malignant hyperthermia and congenital neuromuscular disease with uniform type 1 fiber (CNMDU1). Anesth Analg. 2011;113(6):1461–7.

  58. 58.

    Zhou H, Lillis S, Loy RE, Ghassemi F, Rose MR, Norwood F, Mills K, Al-Sarraj S, Lane RJ, Feng L, et al. Multi-minicore disease and atypical periodic paralysis associated with novel mutations in the skeletal muscle ryanodine receptor (RYR1) gene. Neuromuscul Disord. 2010;20(3):166–73.

  59. 59.

    Sato K, Pollock N, Stowell KM. Functional studies of RYR1 mutations in the skeletal muscle ryanodine receptor using human RYR1 complementary DNA. Anesthesiology. 2010;112(6):1350–4.

  60. 60.

    Merritt A, Booms P, Duke A, Sato K, Stowell K, Steele D, Hopkins PM. Functional analysis of the p.Gly3990Val RYR1 variant using a human cDNA clone in HEK293 cells. Br J Anaesth. 2010;105(5):718P–9P.

  61. 61.

    Migita T, Mukaida K, Hamada H, Yasuda T, Haraki T, Nishino I, Murakami N, Kawamoto M. Functional analysis of ryanodine receptor type 1 p.R2508C mutation in exon 47. J Anesth. 2009;23(3):341–6.

  62. 62.

    Migita T. Do Ca2+ channel blockers improve malignant hyperthermia crisis? Eur J Anaesthesiol. 2009;26(45):124.

  63. 63.

    Ghassemi F, Vukcevic M, Xu L, Zhou H, Meissner G, Muntoni F, Jungbluth H, Zorzato F, Treves S. A recessive ryanodine receptor 1 mutation in a CCD patient increases channel activity. Cell Calcium. 2009;45(2):192–7.

  64. 64.

    Jiang D, Chen W, Xiao J, Wang R, Kong H, Jones PP, Zhang L, Fruen B, Chen SR. Reduced threshold for luminal Ca2+ activation of RyR1 underlies a causal mechanism of porcine malignant hyperthermia. J Biol Chem. 2008;283(30):20813–20.

  65. 65.

    Rossi D, De Smet P, Lyfenko A, Galli L, Lorenzini S, Franci D, Petrioli F, Orrico A, Angelini C, Tegazzin V, et al. A truncation in the RYR1 gene associated with central core lesions in skeletal muscle fibres. J Med Genet. 2007;44(2):e67.

  66. 66.

    Lyfenko AD, Ducreux S, Wang Y, Xu L, Zorzato F, Ferreiro A, Meissner G, Treves S, Dirksen RT. Two central core disease (CCD) deletions in the C-terminal region of RYR1 alter muscle excitation-contraction (EC) coupling by distinct mechanisms. Hum Mutat. 2007;28(1):61–8.

  67. 67.

    Zhou H, Yamaguchi N, Xu L, Wang Y, Sewry C, Jungbluth H, Zorzato F, Bertini E, Muntoni F, Meissner G, et al. Characterization of recessive RYR1 mutations in core myopathies. Hum Mol Genet. 2006;15(18):2791–803.

  68. 68.

    Xu L, Wang Y, Gillespie D, Meissner G. Two rings of negative charges in the cytosolic vestibule of type-1 ryanodine receptor modulate ion fluxes. Biophys J. 2006;90(2):443–53.

  69. 69.

    Wang Y, Xu L, Pasek DA, Gillespie D, Meissner G. Probing the role of negatively charged amino acid residues in ion permeation of skeletal muscle ryanodine receptor. Biophys J. 2005;89(1):256–65.

  70. 70.

    Brini M, Manni S, Pierobon N, Du GG, Sharma P, MacLennan DH, Carafoli E. Ca2+ signaling in HEK-293 and skeletal muscle cells expressing recombinant ryanodine receptors harboring malignant hyperthermia and central core disease mutations. J Biol Chem. 2005;280(15):15380–9.

  71. 71.

    Du GG, Khanna VK, Guo X, MacLennan DH. Central core disease mutations R4892W, I4897T and G4898E in the ryanodine receptor isoform 1 reduce the Ca2+ sensitivity and amplitude of Ca2+−dependent Ca2+ release. Biochem J. 2004;382(Pt 2):557–64.

  72. 72.

    Zorzato F, Yamaguchi N, Xu L, Meissner G, Müller CR, Pouliquin P, Muntoni F, Sewry C, Girard T, Treves S. Clinical and functional effects of a deletion in a COOH-terminal lumenal loop of the skeletal muscle ryanodine receptor. Hum Mol Genet. 2003;12(4):379–88.

  73. 73.

    Stange M, Xu L, Balshaw D, Yamaguchi N, Meissner G. Characterization of recombinant skeletal muscle (Ser-2843) and cardiac muscle (Ser-2809) ryanodine receptor phosphorylation mutants. J Biol Chem. 2003;278(51):51693–702.

  74. 74.

    Loke JC, Kraev N, Sharma P, Du G, Patel L, Kraev A, MacLennan DH. Detection of a novel ryanodine receptor subtype 1 mutation (R328W) in a malignant hyperthermia family by sequencing of a leukocyte transcript. Anesthesiology. 2003;99(2):297–302.

  75. 75.

    Yamaguchi N, Xin C, Meissner G. Identification of apocalmodulin and Ca2+−calmodulin regulatory domain in skeletal muscle Ca2+ release channel, ryanodine receptor. J Biol Chem. 2001;276(25):22579–85.

  76. 76.

    Sun J, Xin C, Eu JP, Stamler JS, Meissner G. Cysteine-3635 is responsible for skeletal muscle ryanodine receptor modulation by NO. Proc Natl Acad Sci U S A. 2001;98(20):11158–62.

  77. 77.

    Gaburjakova M, Gaburjakova J, Reiken S, Huang F, Marx SO, Rosemblit N, Marks AR. FKBP12 binding modulates ryanodine receptor channel gating. J Biol Chem. 2001;276(20):16931–5.

  78. 78.

    Du GG, Oyamada H, Khanna VK, MacLennan DH. Mutations to Gly2370, Gly2373 or Gly2375 in malignant hyperthermia domain 2 decrease caffeine and cresol sensitivity of the rabbit skeletal-muscle Ca2+−release channel (ryanodine receptor isoform 1). Biochem J. 2001;360(Pt 1):97–105.

  79. 79.

    Monnier N, Romero NB, Lerale J, Nivoche Y, Qi D, MacLennan DH, Fardeau M, Lunardi J. An autosomal dominant congenital myopathy with cores and rods is associated with a neomutation in the RYR1 gene encoding the skeletal muscle ryanodine receptor. Hum Mol Genet. 2000;9(18):2599–608.

  80. 80.

    Gao L, Balshaw D, Xu L, Tripathy A, Xin C, Meissner G. Evidence for a role of the lumenal M3-M4 loop in skeletal muscle Ca(2+) release channel (ryanodine receptor) activity and conductance. Biophys J. 2000;79(2):828–40.

  81. 81.

    Tong J, McCarthy TV, MacLennan DH. Measurement of resting cytosolic Ca2+ concentrations and Ca2+ store size in HEK-293 cells transfected with malignant hyperthermia or central core disease mutant Ca2+ release channels. J Biol Chem. 1999;274(2):693–702.

  82. 82.

    Lynch PJ, Tong J, Lehane M, Mallet A, Giblin L, Heffron JJ, Vaughan P, Zafra G, MacLennan DH, McCarthy TV. A mutation in the transmembrane/luminal domain of the ryanodine receptor is associated with abnormal Ca2+ release channel function and severe central core disease. Proc Natl Acad Sci U S A. 1999;96(7):4164–9.

  83. 83.

    Tong J, Oyamada H, Demaurex N, Grinstein S, McCarthy TV, MacLennan DH. Caffeine and halothane sensitivity of intracellular Ca2+ release is altered by 15 calcium release channel (ryanodine receptor) mutations associated with malignant hyperthermia and/or central core disease. J Biol Chem. 1997;272(42):26332–9.

  84. 84.

    Treves S, Larini F, Menegazzi P, Steinberg TH, Koval M, Vilsen B, Andersen JP, Zorzato F. Alteration of intracellular Ca2+ transients in COS-7 cells transfected with the cDNA encoding skeletal-muscle ryanodine receptor carrying a mutation associated with malignant hyperthermia. Biochem J. 1994;301(Pt 3):661–5.

  85. 85.

    Altafaj X, Cheng W, Esteve E, Urbani J, Grunwald D, Sabatier JM, Coronado R, De Waard M, Ronjat M. Maurocalcine and domain A of the II-III loop of the dihydropyridine receptor Cav 1.1 subunit share common binding sites on the skeletal ryanodine receptor. J Biol Chem. 2005;280(6):4013–6.

  86. 86.

    Lefebvre R, Legrand C, Groom L, Dirksen RT, Jacquemond V. Ca2+ release in muscle fibers expressing R4892W and G4896V type 1 ryanodine receptor disease mutants. PLoS One. 2013;8(1):e54042.

  87. 87.

    Groom L, Muldoon SM, Tang ZZ, Brandom BW, Bayarsaikhan M, Bina S, Lee HS, Qiu X, Sambuughin N, Dirksen RT. Identical de novo mutation in the type 1 ryanodine receptor gene associated with fatal, stress-induced malignant hyperthermia in two unrelated families. Anesthesiology. 2011;115(5):938–45.

  88. 88.

    Booms P, Duke AM, Steele D, Shaw MA, Carpenter D, Robinson RL, Halsall PJ, Allen PD, Yang T, Iles DE, et al. Concentration dependence of caffeine-induced Ca2+ release in dyspedic skeletal myotubes transfected with ryanodine receptor isoform-1 (RYR1) cDNAs. Br J Anaesth. 2009;103(2):315P.

  89. 89.

    Yang T, Esteve E, Pessah IN, Molinski TF, Allen PD, Lopez JR. Elevated resting [ca(2+)](i) in myotubes expressing malignant hyperthermia RyR1 cDNAs is partially restored by modulation of passive calcium leak from the SR. Am J Phys Cell Phys. 2007;292(5):C1591–8.

  90. 90.

    Yang T, Allen PD, Pessah IN, Lopez JR. Enhanced excitation-coupled calcium entry in myotubes is associated with expression of RyR1 malignant hyperthermia mutations. J Biol Chem. 2007;282(52):37471–8.

  91. 91.

    Goonasekera SA, Beard NA, Groom L, Kimura T, Lyfenko AD, Rosenfeld A, Marty I, Dulhunty AF, Dirksen RT. Triadin binding to the C-terminal luminal loop of the ryanodine receptor is important for skeletal muscle excitation contraction coupling. J Gen Physiol. 2007;130(4):365–78.

  92. 92.

    Lee EH, Song DW, Lee JM, Meissner G, Allen PD, Kim DH. Occurrence of atypical Ca2+ transients in triadin-binding deficient-RYR1 mutants. Biochem Biophys Res Commun. 2006;351(4):909–14.

  93. 93.

    Aracena-Parks P, Goonasekera SA, Gilman CP, Dirksen RT, Hidalgo C, Hamilton SL. Identification of cysteines involved in S-nitrosylation, S-glutathionylation, and oxidation to disulfides in ryanodine receptor type 1. J Biol Chem. 2006;281(52):40354–68.

  94. 94.

    Hurne AM, O'Brien JJ, Wingrove D, Cherednichenko G, Allen PD, Beam KG, Pessah IN. Ryanodine receptor type 1 (RyR1) mutations C4958S and C4961S reveal excitation-coupled calcium entry (ECCE) is independent of sarcoplasmic reticulum store depletion. J Biol Chem. 2005;280(44):36994–7004.

  95. 95.

    Cheng W, Altafaj X, Ronjat M, Coronado R. Interaction between the dihydropyridine receptor Ca2+ channel beta-subunit and ryanodine receptor type 1 strengthens excitation-contraction coupling. Proc Natl Acad Sci U S A. 2005;102(52):19225–30.

  96. 96.

    Du GG, Avila G, Sharma P, Khanna VK, Dirksen RT, MacLennan DH. Role of the sequence surrounding predicted transmembrane helix M4 in membrane association and function of the Ca(2+) release channel of skeletal muscle sarcoplasmic reticulum (ryanodine receptor isoform 1). J Biol Chem. 2004;279(36):37566–74.

  97. 97.

    Dirksen RT, Avila G. Distinct effects on Ca2+ handling caused by malignant hyperthermia and central core disease mutations in RyR1. Biophys J. 2004;87(5):3193–204.

  98. 98.

    Zhu X, Ghanta J, Walker JW, Allen PD, Valdivia HH. The calmodulin binding region of the skeletal ryanodine receptor acts as a self-modulatory domain. Cell Calcium. 2004;35(2):165–77.

  99. 99.

    Yang T, Ta TA, Pessah IN, Allen PD. Functional defects in six ryanodine receptor isoform-1 (RyR1) mutations associated with malignant hyperthermia and their impact on skeletal excitation-contraction coupling. J Biol Chem. 2003;278(28):25722–30.

  100. 100.

    Avila G, O'Connell KM, Dirksen RT. The pore region of the skeletal muscle ryanodine receptor is a primary locus for excitation-contraction uncoupling in central core disease. J Gen Physiol. 2003;121(4):277–86.

  101. 101.

    Avila G, Lee EH, Perez CF, Allen PD, Dirksen RT. FKBP12 binding to RyR1 modulates excitation-contraction coupling in mouse skeletal myotubes. J Biol Chem. 2003;278(25):22600–8.

  102. 102.

    O'Connell KM, Yamaguchi N, Meissner G, Dirksen RT. Calmodulin binding to the 3614-3643 region of RyR1 is not essential for excitation-contraction coupling in skeletal myotubes. J Gen Physiol. 2002;120(3):337–47.

  103. 103.

    O'Brien JJ, Feng W, Allen PD, Chen SR, Pessah IN, Beam KG. Ca2+ activation of RyR1 is not necessary for the initiation of skeletal-type excitation-contraction coupling. Biophys J. 2002;82(5):2428–35.

  104. 104.

    Feng W, Tu J, Yang T, Vernon PS, Allen PD, Worley PF, Pessah IN. Homer regulates gain of ryanodine receptor type 1 channel complex. J Biol Chem. 2002;277(47):44722–30.

  105. 105.

    Fessenden JD, Chen L, Wang Y, Paolini C, Franzini-Armstrong C, Allen PD, Pessah IN. Ryanodine receptor point mutant E4032A reveals an allosteric interaction with ryanodine. Proc Natl Acad Sci U S A. 2001;98(5):2865–70.

  106. 106.

    Avila G, O'Connell KM, Groom LA, Dirksen RT. Ca2+ release through ryanodine receptors regulates skeletal muscle L-type Ca2+ channel expression. J Biol Chem. 2001;276(21):17732–8.

  107. 107.

    Avila G, O'Brien JJ, Dirksen RT. Excitation--contraction uncoupling by a human central core disease mutation in the ryanodine receptor. Proc Natl Acad Sci U S A. 2001;98(7):4215–20.

  108. 108.

    Avila G, Dirksen RT. Functional effects of central core disease mutations in the cytoplasmic region of the skeletal muscle ryanodine receptor. J Gen Physiol. 2001;118(3):277–90.

  109. 109.

    Moore RA, Nguyen H, Galceran J, Pessah IN, Allen PD. A transgenic myogenic cell line lacking ryanodine receptor protein for homologous expression studies: reconstitution of Ry1R protein and function. J Cell Biol. 1998;140(4):843–51.

  110. 110.

    Zullo A, Perrotta G, D'Angelo R, Ruggiero L, Gravino E, Del Vecchio L, Santoro L, Salvatore F, Carsana A. RYR1 sequence variants in myopathies: expression and functional studies in two families. Biomed Res Int. 2019;2019:7638946.

  111. 111.

    Johannsen S, Treves S, Muller CR, Mogele S, Schneiderbanger D, Roewer N, Schuster F. Functional characterization of the RYR1 mutation p.Arg4737Trp associated with susceptibility to malignant hyperthermia. Neuromuscul Disord. 2016;26(1):21–5.

  112. 112.

    Schiemann AH, Paul N, Parker R, Pollock N, Bulger TF, Stowell KM. Functional characterization of 2 known ryanodine receptor mutations causing malignant hyperthermia. Anesth Analg. 2014;118(2):375–80.

  113. 113.

    Attali R, Aharoni S, Treves S, Rokach O, Becker Cohen M, Fellig Y, Straussberg R, Dor T, Daana M, Mitrani-Rosenbaum S, et al. Variable myopathic presentation in a single family with novel skeletal RYR1 mutation. PLoS One. 2013;8(7):e69296.

  114. 114.

    Vukcevic M, Broman M, Islander G, Bodelsson M, Ranklev-Twetman E, Muller CR, Treves S: Functional properties of RYR1 mutations identified in Swedish patients with malignant hyperthermia and central core disease. Anesth Analg 2010, 111(1):185–190.

  115. 115.

    Grievink H, Stowell KM. Allele-specific differences in ryanodine receptor 1 mRNA expression levels may contribute to phenotypic variability in malignant hyperthermia. Orphanet J Rare Dis. 2010;5:10.

  116. 116.

    Zullo A, Klingler W, De Sarno C, Ferrara M, Fortunato G, Perrotta G, Gravino E, Di Noto R, Lehmann-Horn F, Melzer W, et al. Functional characterization of ryanodine receptor (RYR1) sequence variants using a metabolic assay in immortalized B-lymphocytes. Hum Mutat. 2009;30(4):E575–90.

  117. 117.

    Levano S, Vukcevic M, Singer M, Matter A, Treves S, Urwyler A, Girard T. Increasing the number of diagnostic mutations in malignant hyperthermia. Hum Mutat. 2009;30(4):590–8.

  118. 118.

    Anderson AA, Brown RL, Polster B, Pollock N, Stowell KM. Identification and biochemical characterization of a novel ryanodine receptor gene mutation associated with malignant hyperthermia. Anesthesiology. 2008;108(2):208–15.

  119. 119.

    Ducreux S, Zorzato F, Ferreiro A, Jungbluth H, Muntoni F, Monnier N, Muller CR, Treves S. 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. Biochem J. 2006;395(2):259–66.

  120. 120.

    Tilgen N, Zorzato F, Halliger-Keller B, Muntoni F, Sewry C, Palmucci LM, Schneider C, Hauser E, Lehmann-Horn F, Muller CR, et al. Identification of four novel mutations in the C-terminal membrane spanning domain of the ryanodine receptor 1: association with central core disease and alteration of calcium homeostasis. Hum Mol Genet. 2001;10(25):2879–87.

  121. 121.

    Girard T, Cavagna D, Padovan E, Spagnoli G, Urwyler A, Zorzato F, Treves S. B-lymphocytes from malignant hyperthermia-susceptible patients have an increased sensitivity to skeletal muscle ryanodine receptor activators. J Biol Chem. 2001;276(51):48077–82.

  122. 122.

    Hoppe K, Hack G, Lehmann-Horn F, Jurkat-Rott K, Wearing S, Zullo A, Carsana A, Klingler W. Hypermetabolism in B-lymphocytes from malignant hyperthermia susceptible individuals. Sci Rep. 2016;6:33372.

  123. 123.

    Suman M, Sharpe JA, Bentham RB, Kotiadis VN, Menegollo M, Pignataro V, Molgó J, Muntoni F, Duchen MR, Pegoraro E, et al. Inositol trisphosphate receptor-mediated Ca2+ signalling stimulates mitochondrial function and gene expression in core myopathy patients. Hum Mol Genet. 2018;27(13):2367–82.

  124. 124.

    Choi RH, Koenig X, Launikonis BS. Dantrolene requires Mg<sup>2+</sup> to arrest malignant hyperthermia. Proc Natl Acad Sci. 2017;114(18):4811–5.

  125. 125.

    Kaufmann A, Kraft B, Michalek-Sauberer A, Weindlmayr M, Kress HG, Steinboeck F, Weigl LG. Novel double and single ryanodine receptor 1 variants in two Austrian malignant hyperthermia families. Anesth Analg. 2012;114(5):1017–25.

  126. 126.

    Treves S, Vukcevic M, Jeannet P-Y, Levano S, Girard T, Urwyler A, Fischer D, Voit T, Jungbluth H, Lillis S, et al. Enhanced excitation-coupled Ca2+ entry induces nuclear translocation of NFAT and contributes to IL-6 release from myotubes from patients with central core disease. Hum Mol Genet. 2010;20(3):589–600.

  127. 127.

    Kobayashi M, Mukaida K, Migita T, Hamada H, Kawamoto M, Yuge O. Analysis of human cultured myotubes responses mediated by ryanodine receptor 1. Anaesth Intensive Care. 2011;39(2):252–61.

  128. 128.

    Migita T, Mukaida K, Kawamoto M, Kobayashi M, Nishino I, Yuge O. Propofol-induced changes in myoplasmic calcium concentrations in cultured human skeletal muscles from RYR1 mutation carriers. Anaesth Intensive Care. 2007;35(6):894–8.

  129. 129.

    Broman M, Gehrig A, Islander G, Bodelsson M, Ranklev-Twetman E, Rüffert H, Müller CR. Mutation screening of the RYR1-cDNA from peripheral B-lymphocytes in 15 Swedish malignant hyperthermia index cases. Br J Anaesth. 2009;102(5):642–9.

  130. 130.

    Zhou H, Brockington M, Jungbluth H, Monk D, Stanier P, Sewry CA, Moore GE, Muntoni F. Epigenetic allele silencing unveils recessive RYR1 mutations in core myopathies. Am J Hum Genet. 2006;79(5):859–68.

  131. 131.

    Weigl L. 4-Chloro-m-cresol cannot detect malignant hyperthermia equivocal cells in an alternative minimally invasive diagnostic test of malignant hyperthermia susceptibility. Anesth Analg. 2004;99(1):103–7.

  132. 132.

    Wehner M, Rueffert H, Koenig F, Olthoff D. Functional characterization of malignant hyperthermia-associated RyR1 mutations in exon 44, using the human myotube model. Neuromuscul Disord. 2004;14(7):429–37.

  133. 133.

    Ducreux S, Zorzato F, Muller C, Sewry C, Muntoni F, Quinlivan R, Restagno G, Girard T, Treves S. Effect of ryanodine receptor mutations on interleukin-6 release and intracellular calcium homeostasis in human myotubes from malignant hyperthermia-susceptible individuals and patients affected by central core disease. J Biol Chem. 2004;279(42):43838–46.

  134. 134.

    Wehner M, Rueffert H, Koenig F, Olthoff D. Calcium release from sarcoplasmic reticulum is facilitated in human myotubes derived from carriers of the ryanodine receptor type 1 mutations Ile2182Phe and Gly2375Ala. Genet Test. 2003;7(3):203–11.

  135. 135.

    Wehner M, Rueffert H, Koenig F, Meinecke CD, Olthoff D. The Ile2453Thr mutation in the ryanodine receptor gene 1 is associated with facilitated calcium release from sarcoplasmic reticulum by 4-chloro-m-cresol in human myotubes. Cell Calcium. 2003;34(2):163–8.

  136. 136.

    Wehner M, Rueffert H, Koenig F, Neuhaus J, Olthoff D. Increased sensitivity to 4-chloro-m-cresol and caffeine in primary myotubes from malignant hyperthermia susceptible individuals carrying the ryanodine receptor 1 Thr2206Met (C6617T) mutation. Clin Genet. 2002;62(2):135–46.

  137. 137.

    Sei Y, Brandom BW, Bina S, Hosoi E, Gallagher KL, Wyre HW, Pudimat PA, Holman SJ, Venzon DJ, Daly JW, et al. Patients with malignant hyperthermia demonstrate an altered calcium control mechanism in B lymphocytes. Anesthesiology. 2002;97(5):1052–8.

  138. 138.

    Girard T, Treves S, Censier K, Mueller CR, Zorzato F, Urwyler A. Phenotyping malignant hyperthermia susceptibility by measuring halothane-induced changes in myoplasmic calcium concentration in cultured human skeletal muscle cells. Br J Anaesth. 2002;89(4):571–9.

  139. 139.

    Brinkmeier H, Krämer J, Krämer R, Iaizzo PA, Baur C, Lehmann-Horn F, Rüdel R. Malignant hyperthermia causing Gly2435Arg mutation of the ryanodine receptor facilitates ryanodine-induced calcium release in myotubes. Br J Anaesth. 1999;83(6):855–61.

  140. 140.

    Censier K, Urwyler A, Zorzato F, Treves S. Intracellular calcium homeostasis in human primary muscle cells from malignant hyperthermia-susceptible and normal individuals. Effect of overexpression of recombinant wild-type and Arg163Cys mutated ryanodine receptors. J Clin Invest. 1998;101(6):1233–42.

  141. 141.

    Zullo A, Textor M, Elischer P, Mall S, Alt A, Klingler W, Melzer W. Voltage modulates halothane-triggered Ca(2+) release in malignant hyperthermia-susceptible muscle. J Gen Physiol. 2018;150(1):111–25.

  142. 142.

    O-Uchi J, Mishra J, Jhun BS, Sheu S-S. Malignant hyperthermia-associated mutation of RyR1 induces mitochondrial Ca2+ overload in the cardiomyocytes. FASEB J. 2017;31(1_supplement):1080.1085.

  143. 143.

    Abeele FV, Lotteau S, Ducreux S, Dubois C, Monnier N, Hanna A, Gkika D, Romestaing C, Noyer L, Flourakis M, et al. TRPV1 variants impair intracellular Ca2+ signaling and may confer susceptibility to malignant hyperthermia. Genet Med. 2019;21(2):441–50.

  144. 144.

    Michelucci A, Paolini C, Boncompagni S, Canato M, Reggiani C, Protasi F. Strenuous exercise triggers a life-threatening response in mice susceptible to malignant hyperthermia. FASEB J. 2017;31(8):3649–62.

  145. 145.

    Michelucci A, De Marco A, Guarnier FA, Protasi F, Boncompagni S. Antioxidant treatment reduces formation of structural cores and improves muscle function in RYR1(Y522S/WT) mice. Oxidative Med Cell Longev. 2017;2017:6792694.

  146. 146.

    Lopez RJ, Byrne S, Vukcevic M, Sekulic-Jablanovic M, Xu L, Brink M, Alamelu J, Voermans N, Snoeck M, Clement E, et al. An RYR1 mutation associated with malignant hyperthermia is also associated with bleeding abnormalities. Sci Signal. 2016;9(435):ra68.

  147. 147.

    O-Uchi J, Mishra J, Jhun BS, Hurst S, Fu D, Gomez L, Sheu S-S. Malignant hyperthermia-associated mutation of RyR1 induces mitochondrial damages and cellular oxidation in the heart. FASEB J. 2016;30(1_supplement):960.965.

  148. 148.

    O-Uchi J, Porter G, Kang SH, Boncompagni S, Sokolova N, Gross P, Jhun BS, Beutner G, Brookes P, Blaxall B, et al. RyR1 mutation associated with malignant hyperthermia facilitates catecholaminergic stress-included arrhythmia via mitochondrial injury and oxidative stress (893.8). FASEB J. 2014;28(1_supplement):893.898.

  149. 149.

    Yarotskyy V, Protasi F, Dirksen RT. Accelerated activation of SOCE current in myotubes from two mouse models of anesthetic- and heat-induced sudden death. PLoS One. 2013;8(10):e77633.

  150. 150.

    Vukcevic M, Zorzato F, Keck S, Tsakiris DA, Keiser J, Maizels RM, Treves S. Gain of function in the immune system caused by a ryanodine receptor 1 mutation. J Cell Sci. 2013;126(Pt 15):3485–92.

  151. 151.

    Manno C, Figueroa L, Royer L, Pouvreau S, Lee CS, Volpe P, Nori A, Zhou J, Meissner G, Hamilton SL, et al. Altered Ca2+ concentration, permeability and buffering in the myofibre Ca2+ store of a mouse model of malignant hyperthermia. J Physiol. 2013;591(18):4439–57.

  152. 152.

    Knoblauch M, Dagnino-Acosta A, Hamilton SL. Mice with RyR1 mutation (Y524S) undergo hypermetabolic response to simvastatin. Skelet Muscle. 2013;3(1):22.

  153. 153.

    Lanner JT, Georgiou DK, Dagnino-Acosta A, Ainbinder A, Cheng Q, Joshi AD, Chen Z, Yarotskyy V, Oakes JM, Lee CS, et al. AICAR prevents heat-induced sudden death in RyR1 mutant mice independent of AMPK activation. Nat Med. 2012;18(2):244–51.

  154. 154.

    O-Uchi J, Porter GA, Kang SH, Boncompagni S, Sokolova N, Gross P, Jhun BS, Beutner G, Brookes P, Blaxall BC, et al. Abstract 370: malignant hyperthermia mutation of RyR1 (Y522S) increases catecholamine-induced cardiac arrhythmia through mitochondrial injury. Circ Res. 2012;111(suppl_1):A370.

  155. 155.

    Loy RE, Lueck JD, Mostajo-Radji MA, Carrell EM, Dirksen RT. Allele-specific gene silencing in two mouse models of autosomal dominant skeletal myopathy. PLoS One. 2012;7(11):e49757.

  156. 156.

    Wei L, Salahura G, Boncompagni S, Kasischke KA, Protasi F, Sheu SS, Dirksen RT. Mitochondrial superoxide flashes: metabolic biomarkers of skeletal muscle activity and disease. FASEB J. 2011;25(9):3068–78.

  157. 157.

    Corona BT, Hamilton SL, Ingalls CP. Effect of prior exercise on thermal sensitivity of malignant hyperthermia-susceptible muscle. Muscle Nerve. 2010;42(2):270–2.

  158. 158.

    Boncompagni S, Rossi AE, Micaroni M, Hamilton SL, Dirksen RT, Franzini-Armstrong C, Protasi F. Characterization and temporal development of cores in a mouse model of malignant hyperthermia. Proc Natl Acad Sci U S A. 2009;106(51):21996–2001.

  159. 159.

    Andronache Z, Hamilton SL, Dirksen RT, Melzer W. A retrograde signal from RyR1 alters DHP receptor inactivation and limits window Ca2+ release in muscle fibers of Y522S RyR1 knock-in mice. Proc Natl Acad Sci U S A. 2009;106(11):4531–6.

  160. 160.

    Durham WJ, Aracena-Parks P, Long C, Rossi AE, Goonasekera SA, Boncompagni S, Galvan DL, Gilman CP, Baker MR, Shirokova N, et al. RyR1 S-nitrosylation underlies environmental heat stroke and sudden death in Y522S RyR1 knockin mice. Cell. 2008;133(1):53–65.

  161. 161.

    Corona BT, Rouviere C, Hamilton SL, Ingalls CP. Eccentric contractions do not induce rhabdomyolysis in malignant hyperthermia susceptible mice. J Appl Physiol. 2008;105(5):1542–53.

  162. 162.

    Chelu MG, Goonasekera SA, Durham WJ, Tang W, Lueck JD, Riehl J, Pessah IN, Zhang P, Bhattacharjee MB, Dirksen RT, et al. Heat- and anesthesia-induced malignant hyperthermia in an RyR1 knock-in mouse. FASEB J. 2006;20(2):329–30.

  163. 163.

    Lee CS, Hanna AD, Wang H, Dagnino-Acosta A, Joshi AD, Knoblauch M, Xia Y, Georgiou DK, Xu J, Long C, et al. A chemical chaperone improves muscle function in mice with a RyR1 mutation. Nat Commun. 2017;8:14659.

  164. 164.

    Zvaritch E, MacLennan DH. Muscle spindles exhibit core lesions and extensive degeneration of intrafusal fibers in the Ryr1(I4895T/wt) mouse model of core myopathy. Biochem Biophys Res Commun. 2015;460(1):34–9.

  165. 165.

    De Crescenzo V, Fogarty KE, Lefkowitz JJ, Bellve KD, Zvaritch E, MacLennan DH, Walsh JV Jr. Type 1 ryanodine receptor knock-in mutation causing central core disease of skeletal muscle also displays a neuronal phenotype. Proc Natl Acad Sci U S A. 2012;109(2):610–5.

  166. 166.

    Loy RE, Orynbayev M, Xu L, Andronache Z, Apostol S, Zvaritch E, MacLennan DH, Meissner G, Melzer W, Dirksen RT. Muscle weakness in Ryr1I4895T/WT knock-in mice as a result of reduced ryanodine receptor Ca2+ ion permeation and release from the sarcoplasmic reticulum. J Gen Physiol. 2011;137(1):43–57.

  167. 167.

    Boncompagni S, Loy RE, Dirksen RT, Franzini-Armstrong C. The I4895T mutation in the type 1 ryanodine receptor induces fiber-type specific alterations in skeletal muscle that mimic premature aging. Aging Cell. 2010;9(6):958–70.

  168. 168.

    Zvaritch E, Kraeva N, Bombardier E, McCloy RA, Depreux F, Holmyard D, Kraev A, Seidman CE, Seidman JG, Tupling AR, et al. Ca2+ dysregulation in Ryr1(I4895T/wt) mice causes congenital myopathy with progressive formation of minicores, cores, and nemaline rods. Proc Natl Acad Sci U S A. 2009;106(51):21813–8.

  169. 169.

    Truong KM, Pessah IN. Comparison of chlorantraniliprole and flubendiamide activity toward wild-type and malignant hyperthermia-susceptible ryanodine receptors and heat stress intolerance. Toxicol Sci. 2019;167(2):509–23.

  170. 170.

    Eltit JM, Ding X, Pessah IN, Allen PD, Lopez JR. Nonspecific sarcolemmal cation channels are critical for the pathogenesis of malignant hyperthermia. FASEB J. 2013;27(3):991–1000.

  171. 171.

    Estève E, Eltit J, Bannister R, Liub K, Pessahd I, Beam K, Allen P, Lopez JR. Malignant hyperthermia mutation alters excitation-coupled Ca2+entry in MH RyR1-R163C knock-in myotubes. Fundam Clin Pharmacol. 2010;24(s1):1–106..

  172. 172.

    Giulivi C, Ross-Inta C, Omanska-Klusek A, Napoli E, Sakaguchi D, Barrientos G, Allen PD, Pessah IN. Basal bioenergetic abnormalities in skeletal muscle from ryanodine receptor malignant hyperthermia-susceptible R163C knock-in mice. J Biol Chem. 2011;286(1):99–113.

  173. 173.

    Feng W, Barrientos GC, Cherednichenko G, Yang T, Padilla IT, Truong K, Allen PD, Lopez JR, Pessah IN. Functional and biochemical properties of ryanodine receptor type 1 channels from heterozygous R163C malignant hyperthermia-susceptible mice. Mol Pharmacol. 2011;79(3):420–31.

  174. 174.

    Estève E, Eltit JM, Bannister RA, Liu K, Pessah IN, Beam KG, Allen PD, López JR. A malignant hyperthermia-inducing mutation in RYR1 (R163C): alterations in Ca2+ entry, release, and retrograde signaling to the DHPR. J Gen Physiol. 2010;135(6):619–28.

  175. 175.

    Bannister RA, Estève E, Eltit JM, Pessah IN, Allen PD, López JR, Beam KG. A malignant hyperthermia-inducing mutation in RYR1 (R163C): consequent alterations in the functional properties of DHPR channels. J Gen Physiol. 2010;135(6):629–40.

  176. 176.

    Cherednichenko G, Ward CW, Feng W, Cabrales E, Michaelson L, Samso M, Lopez JR, Allen PD, Pessah IN. Enhanced excitation-coupled calcium entry in myotubes expressing malignant hyperthermia mutation R163C is attenuated by dantrolene. Mol Pharmacol. 2008;73(4):1203–12.

  177. 177.

    Yang T, Riehl J, Esteve E, Matthaei KI, Goth S, Allen PD, Pessah IN, Lopez JR. Pharmacologic and functional characterization of malignant hyperthermia in the R163C RyR1 knock-in mouse. Anesthesiology. 2006;105(6):1164–75.

  178. 178.

    Brennan S, Garcia-Castaneda M, Michelucci A, Sabha N, Malik S, Groom L, Wei LaPierre L, Dowling JJ, Dirksen RT. Mouse model of severe recessive RYR1-related myopathy. Hum Mol Genet. 2019;28(18):3024–36.

  179. 179.

    Elbaz M, Ruiz A, Bachmann C, Eckhardt J, Pelczar P, Venturi E, Lindsay C, Wilson AD, Alhussni A, Humberstone T, et al. Quantitative RyR1 reduction and loss of calcium sensitivity of RyR1Q1970fsX16+A4329D cause cores and loss of muscle strength. Hum Mol Genet. 2019;28(18):2987–99.

  180. 180.

    Elbaz M, Ruiz A, Eckhardt J, Pelczar P, Muntoni F, Boncompagni S, Treves S, Zorzato F. Quantitative reduction of RyR1 protein caused by a single-allele frameshift mutation in RYR1 ex36 impairs the strength of adult skeletal muscle fibres. Hum Mol Genet. 2019;28(11):1872–84.

  181. 181.

    RYR-1 mice. Accessed 17 Dec 2019.

  182. 182.

    Edamame mice. Accessed 17 Dec 2019.

  183. 183.

    Lopez JR, Kaura V, Diggle CP, Hopkins PM, Allen PD. Malignant hyperthermia, environmental heat stress, and intracellular calcium dysregulation in a mouse model expressing the p.G2435R variant of RYR1. Br J Anaesth. 2018;121(4):953–61.

  184. 184.

    Hernandez-Ochoa EO, Melville Z, Vanegas C, Varney KM, Wilder PT, Melzer W, Weber DJ, Schneider MF. Loss of S100A1 expression leads to Ca(2+) release potentiation in mutant mice with disrupted CaM and S100A1 binding to CaMBD2 of RyR1. Phys Rep. 2018;6(15):e13822.

  185. 185.

    Bannister RA, Sheridan DC, Beam KG. Distinct components of retrograde Ca(V)1.1-RyR1 coupling revealed by a lethal mutation in RyR1. Biophys J. 2016;110(4):912–21.

  186. 186.

    Hanson MG, Wilde JJ, Moreno RL, Minic AD, Niswander L. Potassium dependent rescue of a myopathy with core-like structures in mouse. eLife. 2015;4:e02923.

  187. 187.

    Gartz Hanson M, Niswander LA. Rectification of muscle and nerve deficits in paralyzed ryanodine receptor type 1 mutant embryos. Dev Biol. 2015;404(2):76–87.

  188. 188.

    Yuen B, Boncompagni S, Feng W, Yang T, Lopez JR, Matthaei KI, Goth SR, Protasi F, Franzini-Armstrong C, Allen PD, et al. Mice expressing T4826I-RYR1 are viable but exhibit sex- and genotype-dependent susceptibility to malignant hyperthermia and muscle damage. FASEB J. 2012;26(3):1311–22.

  189. 189.

    Barrientos GC, Feng W, Truong K, Matthaei KI, Yang T, Allen PD, Lopez JR, Pessah IN. Gene dose influences cellular and calcium channel dysregulation in heterozygous and homozygous T4826I-RYR1 malignant hyperthermia-susceptible muscle. J Biol Chem. 2012;287(4):2863–76.

  190. 190.

    Andersson DC, Betzenhauser MJ, Reiken S, Umanskaya A, Shiomi T, Marks AR. Stress-induced increase in skeletal muscle force requires protein kinase A phosphorylation of the ryanodine receptor. J Physiol. 2012;590(24):6381–7.

  191. 191.

    Andersson DC, Betzenhauser MJ, Reiken S, Meli AC, Umanskaya A, Xie W, Shiomi T, Zalk R, Lacampagne A, Marks AR. Ryanodine receptor oxidation causes intracellular calcium leak and muscle weakness in aging. Cell Metab. 2011;14(2):196–207.

  192. 192.

    Yamaguchi N, Prosser BL, Ghassemi F, Xu L, Pasek DA, Eu JP, Hernandez-Ochoa EO, Cannon BR, Wilder PT, Lovering RM, et al. Modulation of sarcoplasmic reticulum Ca2+ release in skeletal muscle expressing ryanodine receptor impaired in regulation by calmodulin and S100A1. Am J Phys Cell Phys. 2011;300(5):C998–c1012.

  193. 193.

    Felder E, Protasi F, Hirsch R, Franzini-Armstrong C, Allen PD. Morphology and molecular composition of sarcoplasmic reticulum surface junctions in the absence of DHPR and RyR in mouse skeletal muscle. Biophys J. 2002;82(6):3144–9.

  194. 194.

    Filipova D, Henry M, Rotshteyn T, Brunn A, Carstov M, Deckert M, Hescheler J, Sachinidis A, Pfitzer G, Papadopoulos S. Distinct transcriptomic changes in E14.5 mouse skeletal muscle lacking RYR1 or Cav1.1 converge at E18.5. PLoS One. 2018;13(3):e0194428.

  195. 195.

    Filipova D, Walter A, Gaspar J, Brunn A, Deckert M, Sachinidis A, Pfitzer G, Papadopoulos S. Transcriptomic changes during skeletal muscle development in the presence and absence of the type 1 ryanodine receptor (RYR1). Acta Physiol. 2017;219:38–9.

  196. 196.

    Filipova D, Walter AM, Gaspar JA, Brunn A, Linde NF, Ardestani MA, Deckert M, Hescheler J, Pfitzer G, Sachinidis A, et al. Gene profiling of embryonic skeletal muscle lacking type I ryanodine receptor Ca(2+) release channel. Sci Rep. 2016;6:20050.

  197. 197.

    Filipova D, Walter AM, Gasper JA, Brunn A, Deckert M, Sachinidis A, Pfitzer G, Papadopoulos S. Profound changes in the gene expression of skeletal muscle lacking the type 1 ryanodine receptor (RYR1). Acta Physiol. 2016;216:207–8.

  198. 198.

    Filipova D, Walter A, Gaspar JA, Pfitzer G, Sachinidis A, Papadopoulos S. Genomic profiling of ryanodine receptor type 1 (RYR1)-deficient skeletal muscle. Acta Physiol. 2015;213:152.

  199. 199.

    Bhattacharya D, Mehle A, Kamp TJ, Balijepalli RC. Intramolecular ex vivo fluorescence resonance energy transfer (FRET) of dihydropyridine receptor (DHPR) beta1a subunit reveals conformational change induced by RYR1 in mouse skeletal myotubes. PLoS One. 2015;10(6):e0131399.

  200. 200.

    Hanson MG, Niswander LA. An explant muscle model to examine the refinement of the synaptic landscape. J Neurosci Methods. 2014;238:95–104.

  201. 201.

    Komazaki S, Ikemoto T, Takeshima H, Iino M, Endo M, Nakamura H. Morphological abnormalities of adrenal gland and hypertrophy of liver in mutant mice lacking ryanodine receptors. Cell Tissue Res. 1998;294(3):467–73.

  202. 202.

    Barone V, Bertocchini F, Bottinelli R, Protasi F, Allen PD, Franzini Armstrong C, Reggiani C, Sorrentino V. Contractile impairment and structural alterations of skeletal muscles from knockout mice lacking type 1 and type 3 ryanodine receptors. FEBS Lett. 1998;422(2):160–4.

  203. 203.

    Ikemoto T, Komazaki S, Takeshima H, Nishi M, Noda T, Iino M, Endo M. Functional and morphological features of skeletal muscle from mutant mice lacking both type 1 and type 3 ryanodine receptors. J Physiol. 1997;501(Pt 2):305–12.

  204. 204.

    Nakai J, Dirksen RT, Nguyen HT, Pessah IN, Beam KG, Allen PD. Enhanced dihydropyridine receptor channel activity in the presence of ryanodine receptor. Nature. 1996;380(6569):72–5.

  205. 205.

    Yarotskyy V, Dirksen RT. Temperature and RyR1 regulate the activation rate of store-operated Ca(2)+ entry current in myotubes. Biophys J. 2012;103(2):202–11.

  206. 206.

    Cacheux M, Blum A, Sebastien M, Wozny AS, Brocard J, Mamchaoui K, Mouly V, Roux-Buisson N, Rendu J, Monnier N, et al. Functional characterization of a central core disease RyR1 mutation (p.Y4864H) associated with quantitative defect in RyR1 protein. J Neuromuscul Dis. 2015;2(4):421–32.

  207. 207.

    Bannister RA. Dantrolene-induced inhibition of skeletal L-type Ca2+ current requires RyR1 expression. Biomed Res Int. 2013;2013:390493.

  208. 208.

    Bannister RA, Beam KG. The cardiac alpha(1C) subunit can support excitation-triggered Ca2+ entry in dysgenic and dyspedic myotubes. Channels (Austin). 2009;3(4):268–73.

  209. 209.

    Bannister RA, Beam KG. Ryanodine modification of RyR1 retrogradely affects L-type Ca(2+) channel gating in skeletal muscle. J Muscle Res Cell Motil. 2009;30(5–6):217–23.

  210. 210.

    Sheridan DC, Takekura H, Franzini-Armstrong C, Beam KG, Allen PD, Perez CF. Bidirectional signaling between calcium channels of skeletal muscle requires multiple direct and indirect interactions. Proc Natl Acad Sci U S A. 2006;103(52):19760–5.

  211. 211.

    Perez CF, Lopez JR, Allen PD. Expression levels of RyR1 and RyR3 control resting free Ca2+ in skeletal muscle. Am J Phys Cell Phys. 2005;288(3):C640–9.

  212. 212.

    Beutner G, Sharma VK, Lin L, Ryu SY, Dirksen RT, Sheu SS. Type 1 ryanodine receptor in cardiac mitochondria: transducer of excitation-metabolism coupling. Biochim Biophys Acta. 2005;1717(1):1–10.

  213. 213.

    Protasi F, Shtifman A, Julian FJ, Allen PD. All three ryanodine receptor isoforms generate rapid cooling responses in muscle cells. Am J Phys Cell Phys. 2004;286(3):C662–70.

  214. 214.

    Lorenzon NM, Haarmann CS, Norris EE, Papadopoulos S, Beam KG. Metabolic biotinylation as a probe of supramolecular structure of the triad junction in skeletal muscle. J Biol Chem. 2004;279(42):44057–64.

  215. 215.

    Sheridan DC, Carbonneau L, Ahern CA, Nataraj P, Coronado R. Ca2+−dependent excitation-contraction coupling triggered by the heterologous cardiac/brain DHPR beta2a-subunit in skeletal myotubes. Biophys J. 2003;85(6):3739–57.

  216. 216.

    Perez CF, Mukherjee S, Allen PD. Amino acids 1-1,680 of ryanodine receptor type 1 hold critical determinants of skeletal type for excitation-contraction coupling. Role of divergence domain D2. J Biol Chem. 2003;278(41):39644–52.

  217. 217.

    Fessenden JD, Perez CF, Goth S, Pessah IN, Allen PD. Identification of a key determinant of ryanodine receptor type 1 required for activation by 4-chloro-m-cresol. J Biol Chem. 2003;278(31):28727–35.

  218. 218.

    Fessenden JD, Feng W, Pessah IN, Allen PD. Mutational analysis of putative calcium binding motifs within the skeletal ryanodine receptor isoform, RyR1. J Biol Chem. 2004;279(51):53028–35.

  219. 219.

    Kashiyama T, Murayama T, Suzuki E, Allen PD, Ogawa Y. Frog alpha- and beta-ryanodine receptors provide distinct intracellular Ca2+ signals in a myogenic cell line. PLoS One. 2010;5(7):e11526.

  220. 220.

    Ahern CA, Sheridan DC, Cheng W, Mortenson L, Nataraj P, Allen P, De Waard M, Coronado R. Ca2+ current and charge movements in skeletal myotubes promoted by the beta-subunit of the dihydropyridine receptor in the absence of ryanodine receptor type 1. Biophys J. 2003;84(2 Pt 1):942–59.

  221. 221.

    Protasi F, Paolini C, Nakai J, Beam KG, Franzini-Armstrong C, Allen PD. Multiple regions of RyR1 mediate functional and structural interactions with alpha(1S)-dihydropyridine receptors in skeletal muscle. Biophys J. 2002;83(6):3230–44.

  222. 222.

    Proenza C, O'Brien J, Nakai J, Mukherjee S, Allen PD, Beam KG. Identification of a region of RyR1 that participates in allosteric coupling with the alpha(1S) (Ca(V)1.1) II-III loop. J Biol Chem. 2002;277(8):6530–5.

  223. 223.

    Ward CW, Protasi F, Castillo D, Wang Y, Chen SR, Pessah IN, Allen PD, Schneider MF. Type 1 and type 3 ryanodine receptors generate different Ca(2+) release event activity in both intact and permeabilized myotubes. Biophys J. 2001;81(6):3216–30.

  224. 224.

    Kimura T, Nakamori M, Lueck JD, Pouliquin P, Aoike F, Fujimura H, Dirksen RT, Takahashi MP, Dulhunty AF, Sakoda S. Altered mRNA splicing of the skeletal muscle ryanodine receptor and sarcoplasmic/endoplasmic reticulum Ca2+−ATPase in myotonic dystrophy type 1. Hum Mol Genet. 2005;14(15):2189–200.

  225. 225.

    Estrada M, Cardenas C, Liberona JL, Carrasco MA, Mignery GA, Allen PD, Jaimovich E. Calcium transients in 1B5 myotubes lacking ryanodine receptors are related to inositol trisphosphate receptors. J Biol Chem. 2001;276(25):22868–74.

  226. 226.

    Ward CW, Schneider MF, Castillo D, Protasi F, Wang Y, Chen SR, Allen PD. Expression of ryanodine receptor RyR3 produces Ca2+ sparks in dyspedic myotubes. J Physiol. 2000;525(Pt 1):91–103.

  227. 227.

    Protasi F, Takekura H, Wang Y, Chen SR, Meissner G, Allen PD, Franzini-Armstrong C. RYR1 and RYR3 have different roles in the assembly of calcium release units of skeletal muscle. Biophys J. 2000;79(5):2494–508.

  228. 228.

    O'Connell KM, Dirksen RT. Prolonged depolarization promotes fast gating kinetics of L-type Ca2+ channels in mouse skeletal myotubes. J Physiol. 2000;529(Pt 3):647–59.

  229. 229.

    Fessenden JD, Wang Y, Moore RA, Chen SR, Allen PD, Pessah IN. Divergent functional properties of ryanodine receptor types 1 and 3 expressed in a myogenic cell line. Biophys J. 2000;79(5):2509–25.

  230. 230.

    Conklin MW, Ahern CA, Vallejo P, Sorrentino V, Takeshima H, Coronado R. Comparison of Ca(2+) sparks produced independently by two ryanodine receptor isoforms (type 1 or type 3). Biophys J. 2000;78(4):1777–85.

  231. 231.

    Avila G, Dirksen RT. Functional impact of the ryanodine receptor on the skeletal muscle L-type Ca(2+) channel. J Gen Physiol. 2000;115(4):467–80.

  232. 232.

    Takekura H, Franzini-Armstrong C. Correct targeting of dihydropyridine receptors and triadin in dyspedic mouse skeletal muscle in vivo. Dev Dyn. 1999;214(4):372–80.

  233. 233.

    Flucher BE, Conti A, Takeshima H, Sorrentino V. Type 3 and type 1 ryanodine receptors are localized in triads of the same mammalian skeletal muscle fibers. J Cell Biol. 1999;146(3):621–30.

  234. 234.

    Protasi F, Franzini-Armstrong C, Allen PD. Role of ryanodine receptors in the assembly of calcium release units in skeletal muscle. J Cell Biol. 1998;140(4):831–42.

  235. 235.

    Nakai J, Sekiguchi N, Rando TA, Allen PD, Beam KG. Two regions of the ryanodine receptor involved in coupling with L-type Ca2+ channels. J Biol Chem. 1998;273(22):13403–6.

  236. 236.

    Yamazawa T, Takeshima H, Shimuta M, Iino M. A region of the ryanodine receptor critical for excitation-contraction coupling in skeletal muscle. J Biol Chem. 1997;272(13):8161–4.

  237. 237.

    Nakai J, Ogura T, Protasi F, Franzini-Armstrong C, Allen PD, Beam KG. Functional nonequality of the cardiac and skeletal ryanodine receptors. Proc Natl Acad Sci U S A. 1997;94(3):1019–22.

  238. 238.

    Takeshima H, Yamazawa T, Ikemoto T, Takekura H, Nishi M, Noda T, Iino M. Ca(2+)-induced Ca2+ release in myocytes from dyspedic mice lacking the type-1 ryanodine receptor. EMBO J. 1995;14(13):2999–3006.

  239. 239.

    Popovski ZT, Tanaskovska B, Miskoska-Milevska E, Andonov S, Domazetovska S. Associations of biochemical changes and maternal traits with mutation 1843 (C>T) in the RYR1 gene as a common cause for porcine stress syndrome. Balkan J Med Genet. 2017;19(2):75–80.

  240. 240.

    Scheffler TL, Scheffler JM, Park S, Kasten SC, Wu Y, McMillan RP, Hulver MW, Frisard MI, Gerrard DE. Fiber hypertrophy and increased oxidative capacity can occur simultaneously in pig glycolytic skeletal muscle. Am J Phys Cell Phys. 2014;306(4):C354–63.

  241. 241.

    Bina S, Capacchione J, Muldoon S, Bayarsaikhan M, Bunger R. Lymphocyte-based determination of susceptibility to malignant hyperthermia: a pilot study in swine. Anesthesiology. 2010;113(4):917–24.

  242. 242.

    Liang X, Chen K, Fruen B, Hu J, Ma J, Hu X, Parness J. Impaired interaction between skeletal ryanodine receptors in malignant hyperthermia. Integr Biol. 2009;1(8–9):533–9.

  243. 243.

    Ta TA, Pessah IN. Ryanodine receptor type 1 (RyR1) possessing malignant hyperthermia mutation R615C exhibits heightened sensitivity to dysregulation by non-coplanar 2,2′,3,5′,6-pentachlorobiphenyl (PCB 95). Neurotoxicology. 2007;28(4):770–9.

  244. 244.

    Stinckens A, Van den Maagdenberg K, Luyten T, Georges M, De Smet S, Buys N. The RYR1 g.1843C>T mutation is associated with the effect of the IGF2 intron3-g.3072G>A mutation on muscle hypertrophy. Anim Genet. 2007;38(1):67–71.

  245. 245.

    Murayama T, Oba T, Hara H, Wakebe K, Ikemoto N, Ogawa Y. Postulated role of interdomain interaction between regions 1 and 2 within type 1 ryanodine receptor in the pathogenesis of porcine malignant hyperthermia. Biochem J. 2007;402(2):349–57.

  246. 246.

    Gallant EM, Hart J, Eager K, Curtis S, Dulhunty AF. Caffeine sensitivity of native RyR channels from normal and malignant hyperthermic pigs: effects of a DHPR II-III loop peptide. Am J Phys Cell Phys. 2004;286(4):C821–30.

  247. 247.

    Zhao F, Li P, Chen SR, Louis CF, Fruen BR. Dantrolene inhibition of ryanodine receptor Ca2+ release channels. Molecular mechanism and isoform selectivity. J Biol Chem. 2001;276(17):13810–6.

  248. 248.

    Gallant EM, Curtis S, Pace SM, Dulhunty AF. Arg(615) Cys substitution in pig skeletal ryanodine receptors increases activation of single channels by a segment of the skeletal DHPR II-III loop. Biophys J. 2001;80(4):1769–82.

  249. 249.

    Balog EM, Fruen BR, Shomer NH, Louis CF. Divergent effects of the malignant hyperthermia-susceptible Arg(615)-->Cys mutation on the Ca(2+) and Mg(2+) dependence of the RyR1. Biophys J. 2001;81(4):2050–8.

  250. 250.

    Dietze B, Henke J, Eichinger HM, Lehmann-Horn F, Melzer W. Malignant hyperthermia mutation Arg615Cys in the porcine ryanodine receptor alters voltage dependence of Ca2+ release, 526. J Physiol. 2000;(Pt 3):507–14.

  251. 251.

    Laver DR, Owen VJ, Junankar PR, Taske NL, Dulhunty AF, Lamb GD. Reduced inhibitory effect of Mg2+ on ryanodine receptor-Ca2+ release channels in malignant hyperthermia. Biophys J. 1997;73(4):1913–24.

  252. 252.

    Fruen BR, Mickelson JR, Louis CF. Dantrolene inhibition of sarcoplasmic reticulum Ca2+ release by direct and specific action at skeletal muscle ryanodine receptors. J Biol Chem. 1997;272(43):26965–71.

  253. 253.

    Bašić I, Tadić Z, Lacković V, Gomerčić A. Stress syndrome: ryanodine receptor (RYR1) gene in malignant hyperthermia in humans and pigs. Period Biol. 1997;99(3):313–7.

  254. 254.

    O'Driscoll S, McCarthy TV, Eichinger HM, Erhardt W, Lehmann-Horn F, Herrmann-Frank A. Calmodulin sensitivity of the sarcoplasmic reticulum ryanodine receptor from normal and malignant-hyperthermia-susceptible muscle. Biochem J. 1996;319(Pt 2):421–6.

  255. 255.

    Herrmann-Frank A, Richter M, Lehmann-Horn F. 4-Chloro-m-cresol: a specific tool to distinguish between malignant hyperthermia-susceptible and normal muscle. Biochem Pharmacol. 1996;52(1):149–55.

  256. 256.

    Vogeli P, Bolt R, Fries R, Stranzinger G. Co-segregation of the malignant hyperthermia and the Arg615-Cys615 mutation in the skeletal muscle calcium release channel protein in five European landrace and Pietrain pig breeds. Anim Genet. 1994;25(Suppl 1):59–66.

  257. 257.

    Ledbetter MW, Preiner JK, Louis CF, Mickelson JR. Tissue distribution of ryanodine receptor isoforms and alleles determined by reverse transcription polymerase chain reaction. J Biol Chem. 1994;269(50):31544–51.

  258. 258.

    Fagerlund T, Ording H, Bendixen D, Berg K. Search for three known mutations in the RYR1 gene in 48 Danish families with malignant hyperthermia. Clin Genet. 1994;46(6):401–4.

  259. 259.

    Otsu K, Phillips MS, Khanna VK, de Leon S, MacLennan DH. Refinement of diagnostic assays for a probable causal mutation for porcine and human malignant hyperthermia. Genomics. 1992;13(3):835–7.

  260. 260.

    Hogan K, Couch F, Powers PA, Gregg RG. A cysteine-for-arginine substitution (R614C) in the human skeletal muscle calcium release channel cosegregates with malignant hyperthermia. Anesth Analg. 1992;75(3):441–8.

  261. 261.

    Otsu K, Khanna VK, Archibald AL, MacLennan DH. Cosegregation of porcine malignant hyperthermia and a probable causal mutation in the skeletal muscle ryanodine receptor gene in backcross families. Genomics. 1991;11(3):744–50.

  262. 262.

    McKinney LC, Butler T, Mullen SP, Klein MG. Characterization of ryanodine receptor-mediated calcium release in human B cells: relevance to diagnostic testing for malignant hyperthermia. Anesthesiology. 2006;104(6):1191–201.

  263. 263.

    Gupta VA, Kuwada JY, Beggs AH. P.4.11 developing therapies for congenital myopathies by high throughput chemical screening in ryanodine receptor 1 mutant zebrafish. Neuromuscul Disord. 2013;23(9):762–3.

  264. 264.

    Dowling JJ, Arbogast S, McEvoy A, Nelson DD, Brooks SV, Kuwada JY, Bonnemann CG, Ferreiro A. Increased oxidative stress and successful antioxidant treatment in a vertebrate model of RYR1 related myopathy. Neuromuscul Disord. 2011;21(9–10):720–1.

  265. 265.

    Dowling JJ, McEvoy A, Arbogast S, Kuwada JY, Ferreiro A. Oxidative stress and RYR1-related myopathies. Neuromuscul Disord. 2010;20(9–10):612.

  266. 266.

    Dowling JJ, McEvoy A, Duncan P, Kuwada JY, Feldman EL. Oxidative stress and antioxidant therapy in a zebrafish model of multi minicore myopathy. Ann Neurol. 2009;66:S133.

  267. 267.

    Hirata H, Watanabe T, Hatakeyama J, Sprague SM, Saint-Amant L, Nagashima A, Cui WW, Zhou W, Kuwada JY. Zebrafish relatively relaxed mutants have a ryanodine receptor defect, show slow swimming and provide a model of multi-minicore disease. Development. 2007;134(15):2771–81.

  268. 268.

    Oyamada H, Oguchi K, Saitoh N, Yamazawa T, Hirose K, Kawana Y, Wakatsuki K, Oguchi K, Tagami M, Hanaoka K, et al. Novel mutations in C-terminal channel region of the ryanodine receptor in malignant hyperthermia patients. Jpn J Pharmacol. 2002;88(2):159–66.

  269. 269.

    Vega AV, Ramos-Mondragón R, Calderón-Rivera A, Zarain-Herzberg A, Avila G. Calcitonin gene-related peptide restores disrupted excitation-contraction coupling in myotubes expressing central core disease mutations in RyR1. J Physiol. 2011;589(Pt 19):4649–69.

  270. 270.

    Lefebvre R, Legrand C, González-Rodríguez E, Groom L, Dirksen RT, Jacquemond V. Defects in Ca2+ release associated with local expression of pathological ryanodine receptors in mouse muscle fibres. J Physiol. 2011;589(Pt 22):5361–82.

  271. 271.

    Douris V, Papapostolou KM, Ilias A, Roditakis E, Kounadi S, Riga M, Nauen R, Vontas J. Investigation of the contribution of RyR target-site mutations in diamide resistance by CRISPR/Cas9 genome modification in Drosophila. Insect Biochem Mol Biol. 2017;87:127–35.

  272. 272.

    Gao S, Sandstrom DJ, Smith HE, High B, Marsh JW, Nash HA. Drosophila ryanodine receptors mediate general anesthesia by halothane. Anesthesiology. 2013;118(3):587–601.

  273. 273.

    Sullivan KM, Scott K, Zuker CS, Rubin GM. The ryanodine receptor is essential for larval development in Drosophila melanogaster. Proc Natl Acad Sci U S A. 2000;97(11):5942–7.

  274. 274.

    Wilberger MS, McKenzie EC, Payton ME, Rigas JD, Valberg SJ. Prevalence of exertional rhabdomyolysis in endurance horses in the Pacific Northwestern United States. Equine Vet J. 2015;47(2):165–70.

  275. 275.

    Nieto JE, Aleman M. A rapid detection method for the ryanodine receptor 1 (C7360G) mutation in quarter horses. J Vet Intern Med. 2009;23(3):619–22.

  276. 276.

    Aleman M, Nieto JE, Magdesian KG. Malignant hyperthermia associated with ryanodine receptor 1 (C7360G) mutation in quarter horses. J Vet Intern Med. 2009;23(2):329–34.

  277. 277.

    Aleman M, Riehl J, Aldridge BM, Lecouteur RA, Stott JL, Pessah IN. Association of a mutation in the ryanodine receptor 1 gene with equine malignant hyperthermia. Muscle Nerve. 2004;30(3):356–65.

  278. 278.

    Roberts MC, Mickelson JR, Patterson EE, Nelson TE, Armstrong PJ, Brunson DB, Hogan K. Autosomal dominant canine malignant hyperthermia is caused by a mutation in the gene encoding the skeletal muscle calcium release channel (RYR1). Anesthesiology. 2001;95(3):716–25.

  279. 279.

    Nicoll Baines K, Ferreira C, Hopkins PM, Shaw MA, Hope IA. Aging effects of caenorhabditis elegans ryanodine receptor variants corresponding to human myopathic mutations. G3. 2017;7(5):1451–61.

  280. 280.

    Baines KN, Shaw M-A, Hope IA. Caenorhabditis elegans as a model organism for RYR1 variants and muscle ageing. BMC Anesthesiol. 2014;14(Suppl 1):A21.

  281. 281.

    Hamada T, Sakube Y, Ahnn J, Kim DH, Kagawa H. Molecular dissection, tissue localization and Ca2+ binding of the ryanodine receptor of Caenorhabditis elegans. J Mol Biol. 2002;324(1):123–35.

  282. 282.

    Maryon EB, Saari B, Anderson P. Muscle-specific functions of ryanodine receptor channels in Caenorhabditis elegans. J Cell Sci. 1998;111(Pt 19):2885–95.

  283. 283.

    Sakube Y, Ando H, Kagawa H. An abnormal ketamine response in mutants defective in the ryanodine receptor gene ryr-1(unc-68) of Caenorhabditis elegans11Edited by J. Karn. J Mol Biol. 1997;267(4):849–64.

  284. 284.

    Maryon EB, Coronado R, Anderson P. unc-68 encodes a ryanodine receptor involved in regulating C. elegans body-wall muscle contraction. J Cell Biol. 1996;134(4):885–93.

  285. 285.

    Airey JA, Baring MD, Beck CF, Chelliah Y, Deerinck TJ, Ellisman MH, Houenou LJ, McKemy DD, Sutko JL, Talvenheimo J. Failure to make normal alpha ryanodine receptor is an early event associated with the crooked neck dwarf (cn) mutation in chicken. Dev Dyn. 1993;197(3):169–88.

  286. 286.

    Airey JA, Deerinck TJ, Ellisman MH, Houenou LJ, Ivanenko A, Kenyon JL, McKemy DD, Sutko JL. Crooked neck dwarf (cn) mutant chicken skeletal muscle cells in low density primary cultures fail to express normal alpha ryanodine receptor and exhibit a partial mutant phenotype. Dev Dyn. 1993;197(3):189–202.

  287. 287.

    Ivanenko A, McKemy DD, Kenyon JL, Airey JA, Sutko JL. Embryonic chicken skeletal muscle cells fail to develop normal excitation-contraction coupling in the absence of the alpha ryanodine receptor. Implications for a two-ryanodine receptor system. J Biol Chem. 1995;270(9):4220–3.

  288. 288.

    Oppenheim RW, Prevette D, Houenou LJ, Pincon-Raymond M, Dimitriadou V, Donevan A, O'Donovan M, Wenner P, McKemy DD, Allen PD. Neuromuscular development in the avian paralytic mutant crooked neck dwarf (cn/cn): further evidence for the role of neuromuscular activity in motoneuron survival. J Comp Neurol. 1997;381(3):353–72.

  289. 289.

    Hopkins PM, Rüffert H, Snoeck MM, Girard T, Glahn KPE, Ellis FR, Müller CR, Urwyler A, on behalf of the European Malignant Hyperthermia G, Bandschapp O, et al. European malignant hyperthermia group guidelines for investigation of malignant hyperthermia susceptibility. BJA. 2015;115(4):531–9.

  290. 290.

    Maffioletti SM, Sarcar S, Henderson ABH, Mannhardt I, Pinton L, Moyle LA, Steele-Stallard H, Cappellari O, Wells KE, Ferrari G, et al. Three-dimensional human iPSC-derived artificial skeletal muscles model muscular dystrophies and enable multilineage tissue engineering. Cell Rep. 2018;23(3):899–908.

  291. 291.

    Maleiner B, Tomasch J, Heher P, Spadiut O, Rünzler D, Fuchs C. The importance of biophysical and biochemical stimuli in dynamic skeletal muscle models. Front Physiol. 2018;9:1130.

  292. 292.

    Fusto A, Moyle LA, Gilbert PM, Pegoraro E. Cored in the act: the use of models to understand core myopathies. Dis Model Mech. 2019;12(12):dmm041368.

  293. 293.

    MacLennan DH, Chen SRW. Chapter 116 - ryanodine receptors. In: Bradshaw RA, Dennis EA, editors. Handbook of cell signaling. 2nd ed. San Diego: Academic Press; 2010. p. 927–35.

  294. 294.

    Bovo E, Dvornikov AV, Mazurek SR, de Tombe PP, Zima AV. Mechanisms of Ca2+ handling in zebrafish ventricular myocytes. Pflugers Arch. 2013;465(12):1775–84.

  295. 295.

    Wilson SW, Brand M, Eisen JS. Patterning the zebrafish central nervous system. In: Solnica-Krezel L, editor. Pattern formation in zebrafish. Berlin, Heidelberg: Springer Berlin Heidelberg; 2002. p. 181–215.

  296. 296.

    Maves L. Recent advances using zebrafish animal models for muscle disease drug discovery. Expert Opin Drug Discovery. 2014;9(9):1033–45.

  297. 297.

    Volpatti J, Endo Y, Groom L, Brennan S, Noche R, Zuercher W, Roy P, Dirksen RT, Dowling JJ. Identification of drug modifiers for RYR1 related myopathy using a multi-species discovery pipeline. bioRxiv. 2019;813097.

  298. 298.

    National Research Council Committee on Cost o, Payment for Animal R. The national academies collection: reports funded by National Institutes of Health. In: Strategies that influence cost containment in animal research facilities. Washington (DC): National Academies Press (US) National Academy of Sciences; 2000.

  299. 299.

    Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, Chen PJ, Wilson C, Newby GA, Raguram A, et al. Search-and-replace genome editing without double-strand breaks or donor DNA. Nature. 2019;576:149.

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The authors would like to thank Dr. Joan Austin, Indiana University School of Nursing, for her review of the manuscript.


This work was funded by the Intramural Research Program of the National Institute of Nursing Research (NINR).