The Role of a Brain-specific Splice Variant of Ryanodine Receptor Type 1

Hayashi, Takahiro; Oyamada, Hideto; Yamazawa, Toshiko; Murayama, Takashi; Matsuoka, Toshifumi; Ohba, Keiichiro; Nakano, Michitaka; Oguchi, Katsuji

doi:10.15369/sujms.22.105

Cited by 1 publication

(1 citation statement)

References 25 publications

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“…The Flp-In TM T-REx TM Systems Invitrogen, Carlsbad, CA, USA was also employed as described by Hayashi et al 19 , but with additional refinement procedures to establish the stable cell lines expressing full-length RyR1. In brief, the Flp-In TREx293 cells Invitrgen were cultured in Dulbecco s modified Eagle s medium Wako Pure Chemicals Industries, Ltd., Osaka, Japan with 10 fetal bovine serum and 100 units of penicillin/mL, 100 µg of streptomycin/mL Wako Pure Chemicals Industries, Ltd. .…”

Section: Establishment Of Stable Cell Lines Expressing Doxycycline-inmentioning

confidence: 99%

Construction and Expression of Ryanodine Receptor Mutants Relevant to Malignant Hyperthermia Patients in Japan

Nakano

Oyamada

Yamazawa

et al. 2014

Showa Univ J Med Sci

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: Malignant hyperthermia MH is a potentially fatal pharmacogenetic disorder triggered by exposure to commonly used volatile anesthetics. Pharmacological and genetic analyses implicated the type 1 of ryanodine receptor RyR1 /Ca 2 release channel as the main candidate gene for causing MH. Genetic diagnosis of MH was proposed to replace conventional methods using biopsied muscle samples that are painful for patients and require skillful diagnosticians to interpret. However, more than 250 RyR1 gene variants have now been reported in MH-susceptible patients, although most have yet to be associated with functional abnormalities using exogenous constructs of these mutants expressed in living cells. To directly compare the pharmacological characteristics of some of the MH-related RyR1 mutants, we have established doxycycline -inducible cell lines expressing two of the uncon rmed rabbit RyR1 mutants, Q156K or R534H corresponding to the Q155K or R533H mutations in human RyR1 reported in MH patients in Japan and a confirmed mutant, R164C RyR1 corresponding to the R163C mutation in human . The caffeine sensitivity of Q156K-expressing cells was remarkably enhanced compared to wild-type RyR1 and similarly to previously reported levels for R164C-expressing cells, while that of the R534H mutants was not different from wild-type cells. The resting cytosolic Ca 2 concentrations of cell lines expressing Q156K or R164C were much higher than those expressing R534H or wild-type RyR1. These results indicated that the RyR1 gene mutation causing the Q156K phenotype Q155K in human is potentially susceptible to MH, and that screening for this mutation could be useful for the noninvasive genetic diagnosis of MH in humans.

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Section: Establishment Of Stable Cell Lines Expressing Doxycycline-inmentioning

confidence: 99%