title: Reduced threshold for luminal Ca2+ activation of RyR1 underlies a causal mechanism of porcine malignant hyperthermia.
abstract: Naturally occurring mutations in the skeletal muscle Ca(2+) release channel/ryanodine receptor RyR1 are linked to malignant hyperthermia (MH), a life-threatening complication of general anesthesia. Although it has long been recognized that MH results from uncontrolled or spontaneous Ca(2+) release from the sarcoplasmic reticulum, how MH RyR1 mutations render the sarcoplasmic reticulum susceptible to volatile anesthetic-induced spontaneous Ca(2+) release is unclear. Here we investigated the impact of the porcine MH mutation, R615C, the human equivalent of which also causes MH, on the intrinsic properties of the RyR1 channel and the propensity for spontaneous Ca(2+) release during store Ca(2+) overload, a process we refer to as store overload-induced Ca(2+) release (SOICR). Single channel analyses revealed that the R615C mutation markedly enhanced the luminal Ca(2+) activation of RyR1. Moreover, HEK293 cells expressing the R615C mutant displayed a reduced threshold for SOICR compared with cells expressing wild type RyR1. Furthermore, the MH-triggering agent, halothane, potentiated the response of RyR1 to luminal Ca(2+) and SOICR. Conversely, dantrolene, an effective treatment for MH, suppressed SOICR in HEK293 cells expressing the R615C mutant, but not in cells expressing an RyR2 mutant. These data suggest that the R615C mutation confers MH susceptibility by reducing the threshold for luminal Ca(2+) activation and SOICR, whereas volatile anesthetics trigger MH by further reducing the threshold, and dantrolene suppresses MH by increasing the SOICR threshold. Together, our data support a view in which altered luminal Ca(2+) regulation of RyR1 represents a primary causal mechanism of MH.
authors.affiliation: Libin Cardiovascular Institutes of Alberta, Department of Physiology, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Dawei
authors.initials: D
authors.lastname: Jiang
authors.forename: Wenqian
authors.initials: W
authors.lastname: Chen
authors.forename: Jianmin
authors.initials: J
authors.lastname: Xiao
authors.forename: Ruiwu
authors.initials: R
authors.lastname: Wang
authors.forename: Huihui
authors.initials: H
authors.lastname: Kong
authors.forename: Peter P
authors.initials: PP
authors.lastname: Jones
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.forename: Bradley
authors.initials: B
authors.lastname: Fruen
authors.forename: S R Wayne
authors.initials: SR
authors.lastname: Chen
mesh_terms: D018685:Anesthetics, Inhalation / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D003600:Cytosol / Q000378:metabolism; D003620:Dantrolene / Q000494:pharmacology; D005334:Fever; D006221:Halothane / Q000494:pharmacology; D006801:Humans; D008305:Malignant Hyperthermia / Q000378:metabolism*; D009125:Muscle Relaxants, Central / Q000494:pharmacology; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D013552:Swine
chemical_list: D018685:Anesthetics, Inhalation; D009125:Muscle Relaxants, Central; D003620:Dantrolene; D002118:Calcium; D006221:Halothane