Identification and Biochemical Characterization of a Novel Ryanodine Receptor Gene Mutation Associated with Malignant Hyperthermia

Anderson, Ayuk A.; Brown, Rosemary L.; Polster, Brenda J.; Pollock, Neil; Stowell, Kathryn M.

doi:10.1097/01.anes.0000299431.81267.3e

Cited by 29 publications

(16 citation statements)

References 32 publications

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“…As H4833Y myoblasts were not available, LCLs were used for mRNA stability assays using the transcription inhibitor actinomycin D. LCLs are progressively being used to assess the role of RyR1 mutations in calcium release [11,13]. Possible allelic variations in RyR1 expression levels in LCLs can greatly affect the functional characterization of potentially causative MH mutations.…”

Section: Discussionmentioning

confidence: 99%

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Allele-specific differences in ryanodine receptor 1 mRNA expression levels may contribute to phenotypic variability in malignant hyperthermia

Grievink

Stowell

2010

Orphanet J Rare Dis

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BackgroundMalignant hyperthermia (MH) is a dominantly inherited skeletal muscle disorder that can cause a fatal hypermetabolic reaction to general anaesthetics. The primary locus of MH (MHS1 locus) in humans is linked to chromosome 19q13.1, the position of the gene encoding the ryanodine receptor skeletal muscle calcium release channel (RyR1).MethodsIn this study, an inexpensive allele-specific PCR (AS-PCR) assay was designed that allowed the relative quantification of the two RyR1 transcripts in heterozygous samples found to be susceptible to MH (MHS). Allele-specific differences in RyR1 expression levels can provide insight into the observed variable penetrance and variations in MH phenotypes between individuals. The presence/absence of the H4833Y mutation in RYR1 transcripts was employed as a marker that allowed discrimination between the two alleles.ResultsIn four skeletal muscle samples and two lymphoblastoid cell lines (LCLs) from different MHS patients, the wild type allele was found to be expressed at higher levels than the mutant RyR1 allele. For both LCLs, the ratios between the wild type and mutant RYR1 alleles did not change after different incubation times with actinomycin D. This suggests that there are no allele-specific differences in RyR1 mRNA stability, at least in these cells.ConclusionThe data presented here revealed for the first time allele-specific differences in RYR1 mRNA expression levels in heterozygous MHS samples, and can at least in part contribute to the observed variable penetrance and variations in MH clinical phenotypes.

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Section: Discussionmentioning

confidence: 99%

“…The causative H4833Y MH mutation was used as a marker to allow discrimination between the two RYR 1 transcripts [11]. Measuring the expression of each of the two alleles simultaneously in the one target tissue is optimal for detecting cis -acting differences as each allele serves as an internal control for the other.…”

Section: Introductionmentioning

confidence: 99%

Allele-specific differences in ryanodine receptor 1 mRNA expression levels may contribute to phenotypic variability in malignant hyperthermia

Grievink

Stowell

2010

Orphanet J Rare Dis

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“…If a genetic variant is found, this may be helpful to relatives because they can then have a focused examination of that exon as the first step in evaluating their risk of malignant hyperthermia. However, novel RYR1 variants are often found [7,8 ]. Considerable work is required to demonstrate that a novel variant is indeed a cause of malignant hyperthermia [7,8 ].…”

Section: Evaluation Of Malignant Hyperthermia Susceptibility Preoperamentioning

confidence: 99%

“…However, novel RYR1 variants are often found [7,8 ]. Considerable work is required to demonstrate that a novel variant is indeed a cause of malignant hyperthermia [7,8 ]. But if we could know the genetic risks that our patients bring with them we would be able to deliver the most efficient and appropriate anesthetic care in the ambulatory surgery environment.…”

Section: Evaluation Of Malignant Hyperthermia Susceptibility Preoperamentioning

confidence: 99%

Ambulatory surgery and malignant hyperthermia

Brandom¹

2009

Current Opinion in Anaesthesiology

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Because malignant hyperthermia is potentially lethal, families should be evaluated as thoroughly as possible when an individual who claims to be malignant hyperthermia susceptible presents for elective surgery. Genetic testing may facilitate the evaluation of families with a very strong history of malignant hyperthermia. The use of activated charcoal can speed the removal of potent inhalation anesthetics from anesthesia workstations. This should facilitate the anesthetic care of malignant hyperthermia susceptible patients.

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“…Thus far, more than 200 sequence variants have been identified in the RYR1 gene [Anderson et al, 2008;von der Hagen et al, 2008;Monnier et al, 2008;Sato et al, 2008;Kossugue et al, 2007;Lyfenko et al, 2007;Rossi et al, 2007;Zhou et al, 2007Zhou et al, , 2006Robinson et al, 2006;Wu et al, 2006]. Identification of novel RYR1 variants and their functional characterization help shed light on the molecular bases of the distinct pathophysiological characteristics of each disorderP P(drug-dependent hyperactivity in MH versus muscle weakness and core development in CCD and minicores in MmDP P [Treves et al, 2008]), and are an aid to the diagnosis of MH.…”

Section: Introductionmentioning

confidence: 99%

Functional characterization of ryanodine receptor (RYR1) sequence variants using a metabolic assay in immortalized B-lymphocytes

et al. 2009

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Mutations in the RYR1 gene are linked to malignant hyperthermia (MH), central core disease and multi-minicore disease. We screened by DHPLC the RYR1 gene in 24 subjects for mutations, and characterized functional alterations caused by some RYR1 variants. Three novel sequence variants and twenty novel polymorphisms were identified. Immortalized lymphoblastoid cell lines from patients with RYR1 variants and from controls were stimulated with 4-chloro-mcresol (4-CmC) and the rate of extracellular acidification was recorded. We demonstrate that the increased acidification rate of lymphoblastoid cells in response to 4-CmC is mainly due to RYR1 activation. Cells expressing RYR1 variants in the N-terminal and in the central region of the protein (p.Arg530His, p.Arg2163Pro, p.Asn2342Ser, p.Glu2371Gly and p.Arg2454His) displayed higher activity compared with controls; this could account for the MH-susceptible phenotype. Cell lines harboring RYR1P Cys4664Arg P were significantly less activated by 4-CmC. This result indicates that the p.Cys4664Arg variant causes a leaky channel and depletion of intracellular stores. The functional changes detected corroborate the variants analyzed as disease-causing alterations and the acidification rate measurements as a means to monitor CaP 2+ P-induced metabolic changes in cells harboring mutant RYR1 channels.

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Identification and Biochemical Characterization of a Novel Ryanodine Receptor Gene Mutation Associated with Malignant Hyperthermia

Abstract: DNA analysis to detect mutations which cosegregate with MH as well as biochemical assays on cultured lymphocytes obtained from blood can serve as useful diagnostic tools for MH susceptibility and genotype-phenotype correlations.

Cited by 29 publications

References 32 publications

Allele-specific differences in ryanodine receptor 1 mRNA expression levels may contribute to phenotypic variability in malignant hyperthermia

Allele-specific differences in ryanodine receptor 1 mRNA expression levels may contribute to phenotypic variability in malignant hyperthermia

Ambulatory surgery and malignant hyperthermia

Functional characterization of ryanodine receptor (RYR1) sequence variants using a metabolic assay in immortalized B-lymphocytes

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