title: Removal of FKBP12.6 does not alter the conductance and activation of the cardiac ryanodine receptor or the susceptibility to stress-induced ventricular arrhythmias.
abstract: The 12.6-kDa FK506-binding protein (FKBP12.6) is considered to be a key regulator of the cardiac ryanodine receptor (RyR2), but its precise role in RyR2 function is complex and controversial. In the present study we investigated the impact of FKBP12.6 removal on the properties of the RyR2 channel and the propensity for spontaneous Ca(2+) release and the occurrence of ventricular arrhythmias. Single channel recordings in lipid bilayers showed that FK506 treatment of recombinant RyR2 co-expressed with or without FKBP12.6 or native canine RyR2 did not induce long-lived subconductance states. [(3)H]Ryanodine binding studies revealed that coexpression with or without FKBP12.6 or treatment with or without FK506 did not alter the sensitivity of RyR2 to activation by Ca(2+) or caffeine. Furthermore, single cell Ca(2+) imaging analyses demonstrated that HEK293 cells co-expressing RyR2 and FKBP12.6 or expressing RyR2 alone displayed the same propensity for spontaneous Ca(2+) release or store overload-induced Ca(2+) release (SOICR). FK506 increased the amplitude and decreased the frequency of SOICR in HEK293 cells expressing RyR2 with or without FKBP12.6, indicating that the action of FK506 on SOICR is independent of FKBP12.6. As with recombinant RyR2, the conductance and ligand-gating properties of single RyR2 channels from FKBP12.6-null mice were indistinguishable from those of single wild type channels. Moreover, FKBP12.6-null mice did not exhibit enhanced susceptibility to stress-induced ventricular arrhythmias, in contrast to previous reports. Collectively, our results demonstrate that the loss of FKBP12.6 has no significant effect on the conduction and activation of RyR2 or the propensity for spontaneous Ca(2+) release and stress-induced ventricular arrhythmias.
authors.affiliation: Libin Cardiovascular Institute of Alberta, Department of Physiology and Biophysics, University of Calgary, Calgary, AB, T2N 4N1, Canada.
authors.forename: Jianmin
authors.initials: J
authors.lastname: Xiao
authors.forename: Xixi
authors.initials: X
authors.lastname: Tian
authors.forename: Peter P
authors.initials: PP
authors.lastname: Jones
authors.forename: Jeff
authors.initials: J
authors.lastname: Bolstad
authors.forename: Huihui
authors.initials: H
authors.lastname: Kong
authors.forename: Ruiwu
authors.initials: R
authors.lastname: Wang
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.forename: Henry J
authors.initials: HJ
authors.lastname: Duff
authors.forename: Anne M
authors.initials: AM
authors.lastname: Gillis
authors.forename: Sidney
authors.initials: S
authors.lastname: Fleischer
authors.forename: Michael
authors.initials: M
authors.lastname: Kotlikoff
authors.forename: Julio A
authors.initials: JA
authors.lastname: Copello
authors.forename: S R Wayne
authors.initials: SR
authors.lastname: Chen
mesh_terms: D000818:Animals; D001145:Arrhythmias, Cardiac / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D004285:Dogs; D004562:Electrocardiography / Q000379:methods; D006801:Humans; D008051:Lipid Bilayers / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D008954:Models, Biological; D032342:Muscle Cells / Q000378:metabolism; D051381:Rats; D012433:Ryanodine / Q000737:chemistry; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*; D022021:Tacrolimus Binding Proteins / Q000378:metabolism*
chemical_list: D002118:Calcium; D008051:Lipid Bilayers; D012433:Ryanodine; D019837:Ryanodine Receptor Calcium Release Channel; D022021:Tacrolimus Binding Proteins; C509259:FKBP1B protein, mouse; C403484:tacrolimus binding protein 1B