e5b54c9594084bce64c1015c472251ff 272313.pdf 23cc08d1cddf642cfefd64fc6d5c53858670b422 272313.pdf 60d1edaca05abeb6c8e88f1ec38b86ee6c7c2eda6001b7cf2a7784b024db15a3 272313.pdf Title: Impact of R-Carvedilol on 2-Adrenergic Receptor-Mediated Spontaneous Calcium Release in Human Atrial Myocytes Subject: A hallmark of atrial fibrillation is an excess of spontaneous calcium release events, which can be mimicked by 1- or 2-adrenergic stimulation. Because 1-adrenergic receptor blockers (1-blockers) are primarily used in clinical practice, we here examined the impact of 2-adrenergic stimulation on spontaneous calcium release and assessed whether the R- and S-enantiomers of the non-selective - blocker carvedilol could reverse these effects. For this purpose, human atrial myocytes were isolated from patients undergoing cardiovascular surgery and subjected to confocal calcium imaging or immunofluorescent labeling of the ryanodine receptor (RyR2). Interestingly, the 2-adrenergic agonist fenoterol increased the incidence of calcium sparks and waves to levels observed with the non-specific -adrenergic agonist isoproterenol. Moreover, fenoterol increased both the amplitude and duration of the sparks, facilitating their fusion into calcium waves. Subsequent application of the non -blocking R-Carvedilol enantiomer reversed these effects of fenoterol in a dose-dependent manner. R-Carvedilol also reversed the fenoterol-induced phosphorylation of the RyR2 at Ser-2808 dose-dependently, and 1 M of either R- or S-Carvedilol fully reversed the effect of fenoterol. Together, these findings demonstrate that 2-adrenergic stimulation alone stimulates RyR2 phosphorylation at Ser-2808 and spontaneous calcium release maximally, and points to carvedilol as a tool to attenuate the pathological activation of 2-receptors. Keywords: -adrenergic receptor blocker; human atrial myocyte; sarcoplasmic reticulum; calcium spark; arrhythmia; carvedilol Author: Sergi Casabella-Ramón, Verónica Jiménez-Sábado, Carmen Tarifa, Sandra Casellas, Tien Tina Lu, Paloma Izquierdo-Castro, Ignasi Gich, Marcel Jiménez, Antonino Ginel, José M. Guerra, S. R. Wayne Chen, Raul Benítez and Leif Hove-Madsen Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Jul 25 10:21:34 2022 CEST ModDate: Mon Jul 25 10:27:36 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 13 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3045509 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- ZYYJXI+EURB10 Type 1 Builtin yes yes no 10 0 JYPLJJ+VnURWPalladioL-Bold Type 1 Custom yes yes no 14 0 SXQPWE+VnURWPalladioL Type 1 Custom yes yes no 19 0 CAMXTK+EURM10 Type 1 Builtin yes yes no 23 0 RAZTEI+URWPalladioL-Bold Type 1 Custom yes yes no 27 0 JWNMGI+URWPalladioL-Roma Type 1 Custom yes yes no 32 0 BGRDOM+URWPalladioL-Ital Type 1 Custom yes yes no 36 0 HHFREY+EURM7 Type 1 Builtin yes yes no 40 0 EBFBTV+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 58 0 QMDDXF+CMSY10 Type 1 Builtin yes yes no 83 0 CRJHCU+PazoMath-Italic Type 1 Builtin yes yes no 90 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 103 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-04-01 02:30:44 CEST RepresentationInformation: 272313.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-03-31 13:23:42 CEST Size: 3045509 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 295 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Study Population Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Study Protocol Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: In Vitro Drug Testing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Immunofluorescent Labeling Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Confocal Calcium Imaging Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Data Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Children: Item: Title: Impact of 2-Adrenergic Stimulation on Intracellular Calcium Homeostasis in Human Atrial Myocytes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: R-Carvedilol Dose-Dependently Reverses 2-Adrenergic Stimulation of Spontaneous Ca2+ Release Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Impact of R- and S-Carvedilol Enantiomers on 2-Adrenergic Stimulation of RyR Phosphorylation at Ser-2808 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Children: Item: Title: Impact of 2-Adrenergic Stimulation on Intracellular Calcium Homeostasis in Human Atrial Myocytes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Ability of R- and S-Carvedilol Enantiomers to Reverse the Effects of 2-Adrenergic Stimulation on Ca2+ Homeostasis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Info: Title: Impact of R-Carvedilol on 2-Adrenergic Receptor-Mediated Spontaneous Calcium Release in Human Atrial Myocytes Author: Sergi Casabella-Ramón, Verónica Jiménez-Sábado, Carmen Tarifa, Sandra Casellas, Tien Tina Lu, Paloma Izquierdo-Castro, Ignasi Gich, Marcel Jiménez, Antonino Ginel, José M. Guerra, S. R. Wayne Chen, Raul Benítez and Leif Hove-Madsen Subject: A hallmark of atrial fibrillation is an excess of spontaneous calcium release events, which can be mimicked by 1- or 2-adrenergic stimulation. Because 1-adrenergic receptor blockers (1-blockers) are primarily used in clinical practice, we here examined the impact of 2-adrenergic stimulation on spontaneous calcium release and assessed whether the R- and S-enantiomers of the non-selective - blocker carvedilol could reverse these effects. For this purpose, human atrial myocytes were isolated from patients undergoing cardiovascular surgery and subjected to confocal calcium imaging or immunofluorescent labeling of the ryanodine receptor (RyR2). Interestingly, the 2-adrenergic agonist fenoterol increased the incidence of calcium sparks and waves to levels observed with the non-specific -adrenergic agonist isoproterenol. Moreover, fenoterol increased both the amplitude and duration of the sparks, facilitating their fusion into calcium waves. Subsequent application of the non -blocking R-Carvedilol enantiomer reversed these effects of fenoterol in a dose-dependent manner. R-Carvedilol also reversed the fenoterol-induced phosphorylation of the RyR2 at Ser-2808 dose-dependently, and 1 M of either R- or S-Carvedilol fully reversed the effect of fenoterol. Together, these findings demonstrate that 2-adrenergic stimulation alone stimulates RyR2 phosphorylation at Ser-2808 and spontaneous calcium release maximally, and points to carvedilol as a tool to attenuate the pathological activation of 2-receptors. 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