Excessive Accumulation of Ca2 + in Mitochondria of Y522S-RYR1 Knock-in Mice: A Link Between Leak From the Sarcoplasmic Reticulum and Altered Redox State
Mice (Y522S or YS), carrying a mutation of the sarcoplasmic reticulum (SR) Ca2+ release channel of skeletal muscle fibers (ryanodine receptor type-1, RyR1) which causes Ca2+ leak, are a widely accepted and intensively studied model for human malignant hyperthermia (MH) susceptibility. Since the invo...
Main Authors: | , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2019-09-01
|
Series: | Frontiers in Physiology |
Subjects: | |
Online Access: | https://www.frontiersin.org/article/10.3389/fphys.2019.01142/full |
id |
doaj-95fb6d4c1e4747f793e73027fb9100f5 |
---|---|
record_format |
Article |
spelling |
doaj-95fb6d4c1e4747f793e73027fb9100f52020-11-25T00:43:34ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2019-09-011010.3389/fphys.2019.01142446003Excessive Accumulation of Ca2 + in Mitochondria of Y522S-RYR1 Knock-in Mice: A Link Between Leak From the Sarcoplasmic Reticulum and Altered Redox StateMarta Canato0Paola Capitanio1Lina Cancellara2Luigi Leanza3Anna Raffaello4Denis Vecellio Reane5Lorenzo Marcucci6Antonio Michelucci7Antonio Michelucci8Feliciano Protasi9Feliciano Protasi10Carlo Reggiani11Carlo Reggiani12Department of Biomedical Sciences, School of Medicine and Surgery, University of Padova, Padua, ItalyDepartment of Biomedical Sciences, School of Medicine and Surgery, University of Padova, Padua, ItalyDepartment of Biomedical Sciences, School of Medicine and Surgery, University of Padova, Padua, ItalyDepartment of Biology, University of Padova, Padua, ItalyDepartment of Biomedical Sciences, School of Medicine and Surgery, University of Padova, Padua, ItalyDepartment of Biomedical Sciences, School of Medicine and Surgery, University of Padova, Padua, ItalyDepartment of Biomedical Sciences, School of Medicine and Surgery, University of Padova, Padua, ItalyCenter for Advanced Studies and Technology, Università degli Studi “G. d’Annunzio” Chieti–Pescara, Chieti, ItalyDepartment of Medicine and Aging Sciences, Università degli Studi “G. d’Annunzio” Chieti–Pescara, Chieti, ItalyCenter for Advanced Studies and Technology, Università degli Studi “G. d’Annunzio” Chieti–Pescara, Chieti, ItalyDepartment of Medicine and Aging Sciences, Università degli Studi “G. d’Annunzio” Chieti–Pescara, Chieti, ItalyDepartment of Biomedical Sciences, School of Medicine and Surgery, University of Padova, Padua, ItalyInstitute for Kinesiology Research, Science and Research Center of Koper, Koper, SloveniaMice (Y522S or YS), carrying a mutation of the sarcoplasmic reticulum (SR) Ca2+ release channel of skeletal muscle fibers (ryanodine receptor type-1, RyR1) which causes Ca2+ leak, are a widely accepted and intensively studied model for human malignant hyperthermia (MH) susceptibility. Since the involvement of reactive oxygen species (ROS) and of mitochondria in MH crisis has been previously debated, here we sought to determine Ca2+ uptake in mitochondria and its possible link with ROS production in single fibers isolated from flexor digitorum brevis (FDB) of YS mice. We found that Ca2+ concentration in the mitochondrial matrix, as detected with the ratiometric FRET-based 4mtD3cpv probe, was higher in YS than in wild-type (WT) fibers at rest and after Ca2+ release from SR during repetitive electrical stimulation or caffeine administration. Also mitochondrial ROS production associated with contractile activity (detected with Mitosox probe) was much higher in YS fibers than in WT. Importantly, the inhibition of mitochondrial Ca2+ uptake achieved by silencing MCU reduced ROS accumulation in the matrix and Ca2+ release from SR. Finally, inhibition of mitochondrial ROS accumulation using Mitotempo reduced SR Ca2+ release in YS fibers exposed to caffeine. The present results support the view that mitochondria take up larger amounts of Ca2+ in YS than in WT fibers and that mitochondrial ROS production substantially contributes to the increased caffeine-sensitivity and to the enhanced Ca2+ release from SR in YS fibers.https://www.frontiersin.org/article/10.3389/fphys.2019.01142/fullexcitation–contraction couplingmitochondriareactive oxygen speciesryanodine receptormalignant hyperthermia |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Marta Canato Paola Capitanio Lina Cancellara Luigi Leanza Anna Raffaello Denis Vecellio Reane Lorenzo Marcucci Antonio Michelucci Antonio Michelucci Feliciano Protasi Feliciano Protasi Carlo Reggiani Carlo Reggiani |
spellingShingle |
Marta Canato Paola Capitanio Lina Cancellara Luigi Leanza Anna Raffaello Denis Vecellio Reane Lorenzo Marcucci Antonio Michelucci Antonio Michelucci Feliciano Protasi Feliciano Protasi Carlo Reggiani Carlo Reggiani Excessive Accumulation of Ca2 + in Mitochondria of Y522S-RYR1 Knock-in Mice: A Link Between Leak From the Sarcoplasmic Reticulum and Altered Redox State Frontiers in Physiology excitation–contraction coupling mitochondria reactive oxygen species ryanodine receptor malignant hyperthermia |
author_facet |
Marta Canato Paola Capitanio Lina Cancellara Luigi Leanza Anna Raffaello Denis Vecellio Reane Lorenzo Marcucci Antonio Michelucci Antonio Michelucci Feliciano Protasi Feliciano Protasi Carlo Reggiani Carlo Reggiani |
author_sort |
Marta Canato |
title |
Excessive Accumulation of Ca2 + in Mitochondria of Y522S-RYR1 Knock-in Mice: A Link Between Leak From the Sarcoplasmic Reticulum and Altered Redox State |
title_short |
Excessive Accumulation of Ca2 + in Mitochondria of Y522S-RYR1 Knock-in Mice: A Link Between Leak From the Sarcoplasmic Reticulum and Altered Redox State |
title_full |
Excessive Accumulation of Ca2 + in Mitochondria of Y522S-RYR1 Knock-in Mice: A Link Between Leak From the Sarcoplasmic Reticulum and Altered Redox State |
title_fullStr |
Excessive Accumulation of Ca2 + in Mitochondria of Y522S-RYR1 Knock-in Mice: A Link Between Leak From the Sarcoplasmic Reticulum and Altered Redox State |
title_full_unstemmed |
Excessive Accumulation of Ca2 + in Mitochondria of Y522S-RYR1 Knock-in Mice: A Link Between Leak From the Sarcoplasmic Reticulum and Altered Redox State |
title_sort |
excessive accumulation of ca2 + in mitochondria of y522s-ryr1 knock-in mice: a link between leak from the sarcoplasmic reticulum and altered redox state |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Physiology |
issn |
1664-042X |
publishDate |
2019-09-01 |
description |
Mice (Y522S or YS), carrying a mutation of the sarcoplasmic reticulum (SR) Ca2+ release channel of skeletal muscle fibers (ryanodine receptor type-1, RyR1) which causes Ca2+ leak, are a widely accepted and intensively studied model for human malignant hyperthermia (MH) susceptibility. Since the involvement of reactive oxygen species (ROS) and of mitochondria in MH crisis has been previously debated, here we sought to determine Ca2+ uptake in mitochondria and its possible link with ROS production in single fibers isolated from flexor digitorum brevis (FDB) of YS mice. We found that Ca2+ concentration in the mitochondrial matrix, as detected with the ratiometric FRET-based 4mtD3cpv probe, was higher in YS than in wild-type (WT) fibers at rest and after Ca2+ release from SR during repetitive electrical stimulation or caffeine administration. Also mitochondrial ROS production associated with contractile activity (detected with Mitosox probe) was much higher in YS fibers than in WT. Importantly, the inhibition of mitochondrial Ca2+ uptake achieved by silencing MCU reduced ROS accumulation in the matrix and Ca2+ release from SR. Finally, inhibition of mitochondrial ROS accumulation using Mitotempo reduced SR Ca2+ release in YS fibers exposed to caffeine. The present results support the view that mitochondria take up larger amounts of Ca2+ in YS than in WT fibers and that mitochondrial ROS production substantially contributes to the increased caffeine-sensitivity and to the enhanced Ca2+ release from SR in YS fibers. |
topic |
excitation–contraction coupling mitochondria reactive oxygen species ryanodine receptor malignant hyperthermia |
url |
https://www.frontiersin.org/article/10.3389/fphys.2019.01142/full |
work_keys_str_mv |
AT martacanato excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT paolacapitanio excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT linacancellara excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT luigileanza excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT annaraffaello excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT denisvecellioreane excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT lorenzomarcucci excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT antoniomichelucci excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT antoniomichelucci excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT felicianoprotasi excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT felicianoprotasi excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT carloreggiani excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate AT carloreggiani excessiveaccumulationofca2inmitochondriaofy522sryr1knockinmicealinkbetweenleakfromthesarcoplasmicreticulumandalteredredoxstate |
_version_ |
1725277632046563328 |