title: 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.
abstract: More than 80 mutations in the skeletal muscle ryanodine receptor gene have been found to be associated with autosomal dominant forms of malignant hyperthermia and central core disease, and with recessive forms of multi-minicore disease. Studies on the functional effects of pathogenic dominant mutations have shown that they mostly affect intracellular Ca2+ homoeostasis, either by rendering the channel hypersensitive to activation (malignant hyperthermia) or by altering the amount of Ca2+ released subsequent to physiological or pharmacological activation (central core disease). In the present paper, we show, for the first time, data on the functional effect of two recently identified recessive ryanodine receptor 1 amino acid substitutions, P3527S and V4849I, as well as that of R999H, another substitution that was identified in two siblings that were affected by multi-minicore disease. We studied the intracellular Ca2+ homoeostasis of EBV (Epstein-Barr virus)-transformed lymphoblastoid cells from the affected patients, their healthy relatives and control individuals. Our results show that the P3527S substitution in the homozygous state affected the amount of Ca2+ released after pharmacological activation with 4-chloro-m-cresol and caffeine, but did not affect the size of the thapsigargin-sensitive Ca2+ stores. The other substitutions had no effect on either the size of the intracellular Ca2+ stores, or on the amount of Ca2+ released after ryanodine receptor activation; however, both the P3527S and V4849I substitutions had a small but significant effect on the resting Ca2+ concentration.
authors.affiliation: Department of Anaesthesia and Research, Basel University Hospital, 4031 Basel, Switzerland.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Ducreux
authors.forename: Francesco
authors.initials: F
authors.lastname: Zorzato
authors.forename: Ana
authors.initials: A
authors.lastname: Ferreiro
authors.forename: Heinz
authors.initials: H
authors.lastname: Jungbluth
authors.forename: Francesco
authors.initials: F
authors.lastname: Muntoni
authors.forename: Nicole
authors.initials: N
authors.lastname: Monnier
authors.forename: Clemens R
authors.initials: CR
authors.lastname: Müller
authors.forename: Susan
authors.initials: S
authors.lastname: Treves
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics*; D002110:Caffeine / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects; D016022:Case-Control Studies; D002478:Cells, Cultured; D003408:Cresols / Q000493:pharmacokinetics; D004252:DNA Mutational Analysis; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D020512:Myopathy, Central Core / Q000378:metabolism*; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019284:Thapsigargin / Q000494:pharmacology
chemical_list: D003408:Cresols; D019837:Ryanodine Receptor Calcium Release Channel; C006984:chlorocresol; D002110:Caffeine; D019284:Thapsigargin