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Functional Properties of RYR1 Mutations Identified in Swedish Patients with Malignant Hyperthermia and Central Core Disease

Vukcevic, Mirko; Broman, Marcus; Islander, Gunilla LU ; Bodelsson, Mikael LU ; Ranklev Twetman, Eva LU ; Mueller, Clemens R. and Treves, Susan (2010) In Anesthesia and Analgesia 111(1). p.185-190
Abstract
BACKGROUND: A diagnosis of malignant hyperthermia susceptibility by in vitro contraction testing can often only be performed at specialized laboratories far away from where patients live. Therefore, we have designed a protocol for genetic screening of the RYR1-cDNA and for functional testing of newly identified ryanodine receptor 1 (RYR1) gene variants in B lymphocytes isolated from peripheral blood samples drawn at local primary care centers. METHODS: B lymphocytes were isolated for the extraction of RYR1-mRNA and genomic DNA and for establishment of lymphoblastoid B cell lines in 5 patients carrying yet unclassified mutations in the RYR1. The B lymphoblastoid cell lines were used to study resting cytoplasmic calcium concentration, the... (More)
BACKGROUND: A diagnosis of malignant hyperthermia susceptibility by in vitro contraction testing can often only be performed at specialized laboratories far away from where patients live. Therefore, we have designed a protocol for genetic screening of the RYR1-cDNA and for functional testing of newly identified ryanodine receptor 1 (RYR1) gene variants in B lymphocytes isolated from peripheral blood samples drawn at local primary care centers. METHODS: B lymphocytes were isolated for the extraction of RYR1-mRNA and genomic DNA and for establishment of lymphoblastoid B cell lines in 5 patients carrying yet unclassified mutations in the RYR1. The B lymphoblastoid cell lines were used to study resting cytoplasmic calcium concentration, the peak calcium transient induced by the sarco(endo)plasmic reticulum Ca-ATPase inhibitor thapsigargin, and the dose-dependent calcium release induced by the ryanodine receptor agonist 4-chloro-m-cresol. RESULTS: It was possible to extract mRNA for cDNA synthesis and to create B lymphocyte clones from all samples. All B lymphoblastoid cell lines carrying RYR1 candidate mutations showed significantly increased resting cytoplasmic calcium levels as well as a shift to lower concentrations of 4-chloro-m-cresol inducing calcium release compared with controls. CONCLUSIONS: Peripheral blood samples are stable regarding RNA and DNA extraction and establishment of lymphoblastoid B cell lines after transportation at ambient temperature over large distances by ordinary mail. Functional tests on B cells harboring the newly identified amino acid substitutions indicate that they alter intracellular Ca2+ homeostasis and are most likely causative of malignant hyperthermia. (Anesth Analg 2010;111:185-90) (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Anesthesia and Analgesia
volume
111
issue
1
pages
185 - 190
publisher
Lippincott Williams & Wilkins
external identifiers
  • wos:000279281500032
  • scopus:77954684290
ISSN
1526-7598
DOI
10.1213/ANE.0b013e3181cbd815
language
English
LU publication?
yes
id
37d8bf76-2edc-4762-aeea-4aff7a013ae1 (old id 1629389)
alternative location
http://www.anesthesia-analgesia.org/content/111/1/185.abstract
date added to LUP
2010-07-23 11:57:19
date last changed
2018-05-29 10:16:16
@article{37d8bf76-2edc-4762-aeea-4aff7a013ae1,
  abstract     = {BACKGROUND: A diagnosis of malignant hyperthermia susceptibility by in vitro contraction testing can often only be performed at specialized laboratories far away from where patients live. Therefore, we have designed a protocol for genetic screening of the RYR1-cDNA and for functional testing of newly identified ryanodine receptor 1 (RYR1) gene variants in B lymphocytes isolated from peripheral blood samples drawn at local primary care centers. METHODS: B lymphocytes were isolated for the extraction of RYR1-mRNA and genomic DNA and for establishment of lymphoblastoid B cell lines in 5 patients carrying yet unclassified mutations in the RYR1. The B lymphoblastoid cell lines were used to study resting cytoplasmic calcium concentration, the peak calcium transient induced by the sarco(endo)plasmic reticulum Ca-ATPase inhibitor thapsigargin, and the dose-dependent calcium release induced by the ryanodine receptor agonist 4-chloro-m-cresol. RESULTS: It was possible to extract mRNA for cDNA synthesis and to create B lymphocyte clones from all samples. All B lymphoblastoid cell lines carrying RYR1 candidate mutations showed significantly increased resting cytoplasmic calcium levels as well as a shift to lower concentrations of 4-chloro-m-cresol inducing calcium release compared with controls. CONCLUSIONS: Peripheral blood samples are stable regarding RNA and DNA extraction and establishment of lymphoblastoid B cell lines after transportation at ambient temperature over large distances by ordinary mail. Functional tests on B cells harboring the newly identified amino acid substitutions indicate that they alter intracellular Ca2+ homeostasis and are most likely causative of malignant hyperthermia. (Anesth Analg 2010;111:185-90)},
  author       = {Vukcevic, Mirko and Broman, Marcus and Islander, Gunilla and Bodelsson, Mikael and Ranklev Twetman, Eva and Mueller, Clemens R. and Treves, Susan},
  issn         = {1526-7598},
  language     = {eng},
  number       = {1},
  pages        = {185--190},
  publisher    = {Lippincott Williams & Wilkins},
  series       = {Anesthesia and Analgesia},
  title        = {Functional Properties of RYR1 Mutations Identified in Swedish Patients with Malignant Hyperthermia and Central Core Disease},
  url          = {http://dx.doi.org/10.1213/ANE.0b013e3181cbd815},
  volume       = {111},
  year         = {2010},
}