Frog α- and β-Ryanodine Receptors Provide Distinct Intracellular Ca2+ Signals in a Myogenic Cell Line 英文参考文献.docVIP

Froga-andb-RyanodineReceptorsProvideDistinct IntracellularCa SignalsinaMyogenicCellLine 2+ TakuKashiyama1,TakashiMurayama1*,ErikaSuzuki1,PaulD.Allen2,YasuoOgawa1 1Department of Pharmacology, Juntendo University School of Medicine, Tokyo, Japan, 2Department of Anesthesia, Perioperative and Pain Medicine, Brigham and Women’sHospital,Boston,Massachusetts,UnitedStatesofAmerica Abstract Background:Infrogskeletalmuscle,tworyanodinereceptor(RyR)isoforms,a-RyRandb-RyR,areexpressedinnearlyequal amounts.However, therolesandsignificance ofthetwoisoforms inexcitation-contraction (E-C)couplingremains tobe elucidated. Methodology/PrincipalFindings:Inthisstudy,weexpressedeitherorbotha-RyRandb-RyRin1B5RyR-deficientmyotubes usingtheherpessimplexvirus1helper-freeampliconsystem.Immunologicalcharacterizationsrevealedthata-RyRandb- RyR are appropriately expressed and targeted at the junctions in 1B5 myotubes. In Ca imaging studies, each isoform 2+ 2+ 2+ 2+ 2+ exhibitedcaffeine-inducedCa transients,anindicativeofCa -inducedCa release(CICR).However,thefashionofCa release events was fundamentally different: a-RyR mediated graded and sustained Ca release observed uniformly throughoutthecytoplasm,whereasb-RyRsupportedall-or-nonetyperegenerativeCa oscillationsandwaves.a-RyRbut 2+ 2+ notb-RyRexhibitedCa transientstriggeredbymembranedepolarizationwithhigh[K+]othatwerenifedipine-sensitive, indicating that only a-RyR mediates depolarization-induced Ca release. Myotubes co-expressing a-RyR and b-RyR demonstratedhigh[K ]o-inducedCa transientswhichwereindistinguishablefromthosewithmyotubesexpressinga-RyR alone. Furthermore, procaine did not affect the peak height of high [K ]o-induced Ca transients, suggesting minor amplificationofCa releasebyb-RyRviaCICRin1B5myotubes. 2+ 2+ + 2+ + 2+ 2+ Conclusions/Significance: These findings suggest that a-RyR and b-RyR provide distinct intracellular Ca2+ signals in a myogeniccellline.Thesedistinctpropertiesmayalsooccurinfrogskeletalmus