frog α- and β-ryanodine receptors provide distinct intracellular ca2+ signals in a myogenic cell line青蛙α-和β-ryanodine受体提供不同的肌原性的细胞系的细胞内钙离子信号.pdfVIP
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frogα-andβ-ryanodinereceptorsprovidedistinctintracellularca2signalsinamyogeniccellline青蛙α-和β-ryanodine受体提供不同的肌原性的细胞系的细胞内钙离子信号
Frog a- and b-Ryanodine Receptors Provide Distinct Intracellular Ca2+ Signals in a Myogenic Cell Line 1 1 1 2 1 Taku Kashiyama , Takashi Murayama *, Erika Suzuki , Paul D. Allen , Yasuo Ogawa 1 Department of Pharmacology, Juntendo University School of Medicine, Tokyo, Japan, 2 Department of Anesthesia, Perioperative and Pain Medicine, Brigham and Women’s Hospital, Boston, Massachusetts, United States of America Abstract Background: In frog skeletal muscle, two ryanodine receptor (RyR) isoforms, a-RyR and b-RyR, are expressed in nearly equal amounts. However, the roles and significance of the two isoforms in excitation-contraction (E-C) coupling remains to be elucidated. Methodology/Principal Findings: In this study, we expressed either or both a-RyR and b-RyR in 1B5 RyR-deficient myotubes using the herpes simplex virus 1 helper-free amplicon system. Immunological characterizations revealed that a-RyR and b- RyR are appropriately expressed and targeted at the junctions in 1B5 myotubes. In Ca2+ imaging studies, each isoform exhibited caffeine-induced Ca2+ transients, an indicative of Ca2+-induced Ca2+ release (CICR). However, the fashion of Ca2+ release events was fundamentally different: a-RyR mediated graded and sustained Ca2+ release observed uniformly throughout the cytoplasm, whereas b-RyR supported all-or-none type regenerative Ca2+ oscillations and waves. a-RyR but 2+ + not b-RyR exhibited Ca transients triggered by membrane depolarization with high [K ]o that were nifedipine-sensitive, indicating that only a-RyR mediates depolarizatio
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