Skeletal Muscle. Ca Release Control Inside the Sarcoplasmic Reticulum.

Summary

Principal Investigator: Eduardo Rios
Abstract: DESCRIPTION (provided by applicant): Ca2+ signaling is a universal language used by cells to react and change. In skeletal muscle its patterns of interest cover multiple time scales: milliseconds -Ca2+ movements that determine contraction and relaxation-;seconds to minutes -when sustained activity may lead to myogenic fatigue- and hours to weeks -patterns that cause changes in gene expression and long-term adaptation-. This study is about inside its cellular store;its quantity, and its concentration, [Ca2+]SR, which conditions Ca2+ signals in every time scale. We ask (1) whether and how [Ca2+]SR controls Ca2+ release from the store, and (2) whether and how calsequestrin and triadin, two strategically located SR proteins, contribute to this control. A technical task, which we call "aim 0", is to image and measure [Ca2+]SR. This was accomplished in the current period and will continue in the next, using novel biosensors -molecules made by the cells themselves- and new hybrid monitors, consisting of high performance small synthetic sensors placed into cells manipulated to make special bio-anchors. To answer questions 1 and 2, we will respectively manipulate [Ca2+]SR while we measure it (aim 1) and force cells to change their endowment of calsequestrin and triadin (aim 2). These goals are now feasible in living animals thanks to a DNA transfection method that works with every protein and can be used also to prevent their synthesis. We propose that [Ca2+]SR -which decays when muscles fatigue- is sustained by SOCE, a universal Ca2+ entry pathway, crucial for mobilizing transcription factors that control gene expression. Using SOCE measures developed in the first period, we propose as aim 3 to define the role of newly discovered molecules of SOCE in the control of [Ca2+]SR. These molecules could be bulwarks against fatigue, and provide powerful tools for experimental alterations of [Ca2+]SR in iterative approaches to the main questions. Ca2+ signals deteriorate in disease, fatigue and aging. Fast Ca2+ signals fail in diseases like hypo-PP, MH susceptibility and central core and minicore, as well as in ageing muscle. Mid-range signaling is affected in fatigue and in an MH-like phenotype of mice lacking calsequestrin. Diseases of long term Ca2+ signals, which show striking parallels in muscle and the immune system, include SCID, a familial immune defect that combines loss of SOCE in lymphocytes and a myogenic myopathy. Our work will advance understanding of these deficits by evaluating roles of specific molecules and their interactions. While only fatigue will be specifically addressed in the present project, questions on the relationships among deficits of function, the intricate pathophysiology and the rational design of therapeutic corrections will be addressed better as we understand what controls stored calcium, and what the stored calcium controls. PUBLIC HEALTH RELEVANCE: This project and our lab's work deal with movements of calcium inside muscle. So-called "calcium signaling" is a universal language used by cells to react and change. In skeletal muscle these signals make the difference between rest and motion, thus influencing multiple aspects of our life, including physical conditioning, metabolism, and even body weight. Because calcium is used for many tasks, including killing cells, the signals are dangerous and must be kept under control. In this project we ask what controls the movements of calcium, in particular focusing on roles of calcium storage areas inside the cell. We also ask how the intra-store calcium determines how much will move, or when the movement will cease. Therefore, our work should help understand why signals fail, or get out of hand. This occurs in diseases, including muscular dystrophies, malignant hyperthermia, and others. Signals also decay during fatigue -their decay actually causing fatigue-- and in old age. In muscle fatigue, the stores are depleted. We propose that two newly discovered molecules are important factors that delay this depletion. These molecules are known to play roles in organs and cells other than muscle, including cells that combat infections. As a result there will be multiple repercussions to their defects or failures. It can be anticipated, therefore, that our research will have consequences outside its field of muscle and exercise.
Funding Period: 2003-01-01 - 2014-04-30
more information: NIH RePORT

Top Publications

  1. ncbi A probable role of dihydropyridine receptors in repression of Ca2+ sparks demonstrated in cultured mammalian muscle
    Jingsong Zhou
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago 60612, USA
    Am J Physiol Cell Physiol 290:C539-53. 2006
  2. pmc Confocal imaging of transmembrane voltage by SEER of di-8-ANEPPS
    Carlo Manno
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 141:371-87. 2013
  3. pmc Using two dyes with the same fluorophore to monitor cellular calcium concentration in an extended range
    Lourdes Figueroa
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, Illinois, USA
    PLoS ONE 8:e55778. 2013
  4. pmc Dynamic measurement of the calcium buffering properties of the sarcoplasmic reticulum in mouse skeletal muscle
    Carlo Manno
    Section of Cellular Signaling Department of Molecular Biophysics and Physiology, Rush University School of Medicine, 1750 W Harrison St, Chicago, IL 60612, USA
    J Physiol 591:423-42. 2013
  5. pmc Properties of Ca2+ sparks revealed by four-dimensional confocal imaging of cardiac muscle
    Vyacheslav M Shkryl
    Department of Molecular Biophysics and Physiology, Section of Cellular Signaling, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 139:189-207. 2012
  6. pmc Synthetic localized calcium transients directly probe signalling mechanisms in skeletal muscle
    Lourdes Figueroa
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University School of Medicine, 1750 W Harrison St, Suite 1279JS, Chicago, IL 60612, USA
    J Physiol 590:1389-411. 2012
  7. pmc Mitochondrial calcium uptake regulates rapid calcium transients in skeletal muscle during excitation-contraction (E-C) coupling
    Jianxun Yi
    Department of Molecular Biophysics and Physiology, Rush University School of Medicine, Chicago, Illinois 60612, USA
    J Biol Chem 286:32436-43. 2011
  8. pmc Measurement of RyR permeability reveals a role of calsequestrin in termination of SR Ca(2+) release in skeletal muscle
    Monika Sztretye
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 138:231-47. 2011
  9. pmc D4cpv-calsequestrin: a sensitive ratiometric biosensor accurately targeted to the calcium store of skeletal muscle
    Monika Sztretye
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 138:211-29. 2011
  10. pmc Paradoxical buffering of calcium by calsequestrin demonstrated for the calcium store of skeletal muscle
    Leandro Royer
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 136:325-38. 2010

Detail Information

Publications17

  1. ncbi A probable role of dihydropyridine receptors in repression of Ca2+ sparks demonstrated in cultured mammalian muscle
    Jingsong Zhou
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago 60612, USA
    Am J Physiol Cell Physiol 290:C539-53. 2006
    ..These data suggest that DHPRs play a critical role in reducing the rate of spontaneous opening of Ca(2+) release channels and/or their susceptibility to Ca(2+)-induced activation, thereby suppressing the production of Ca(2+) sparks...
  2. pmc Confocal imaging of transmembrane voltage by SEER of di-8-ANEPPS
    Carlo Manno
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 141:371-87. 2013
    ..These images resolved the radially varying lag associated with propagation at a finite velocity...
  3. pmc Using two dyes with the same fluorophore to monitor cellular calcium concentration in an extended range
    Lourdes Figueroa
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, Illinois, USA
    PLoS ONE 8:e55778. 2013
    ..As illustrative application of the enhanced measurement, we use fluo-4 and fluo-4FF to image the calcium wave produced by a cardiac myocyte in response to a small artificial calcium spark...
  4. pmc Dynamic measurement of the calcium buffering properties of the sarcoplasmic reticulum in mouse skeletal muscle
    Carlo Manno
    Section of Cellular Signaling Department of Molecular Biophysics and Physiology, Rush University School of Medicine, 1750 W Harrison St, Chicago, IL 60612, USA
    J Physiol 591:423-42. 2013
    ..The reduction in B during release might reflect changes in calsequestrin conformation upon calcium loss...
  5. pmc Properties of Ca2+ sparks revealed by four-dimensional confocal imaging of cardiac muscle
    Vyacheslav M Shkryl
    Department of Molecular Biophysics and Physiology, Section of Cellular Signaling, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 139:189-207. 2012
    ....
  6. pmc Synthetic localized calcium transients directly probe signalling mechanisms in skeletal muscle
    Lourdes Figueroa
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University School of Medicine, 1750 W Harrison St, Suite 1279JS, Chicago, IL 60612, USA
    J Physiol 590:1389-411. 2012
    ..The differences in flux and threshold may be ascribed to the absence of ryanodine receptor 3 (RyR3) isoforms in adult mammalian muscle...
  7. pmc Mitochondrial calcium uptake regulates rapid calcium transients in skeletal muscle during excitation-contraction (E-C) coupling
    Jianxun Yi
    Department of Molecular Biophysics and Physiology, Rush University School of Medicine, Chicago, Illinois 60612, USA
    J Biol Chem 286:32436-43. 2011
    ....
  8. pmc Measurement of RyR permeability reveals a role of calsequestrin in termination of SR Ca(2+) release in skeletal muscle
    Monika Sztretye
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 138:231-47. 2011
    ..This observation indicates that [Ca(2+)](SR), sensed by Casq and transmitted to the channels presumably via connecting proteins, is determinant to cause the closure that terminates Ca(2+) release...
  9. pmc D4cpv-calsequestrin: a sensitive ratiometric biosensor accurately targeted to the calcium store of skeletal muscle
    Monika Sztretye
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 138:211-29. 2011
    ..It is demonstrably small in D4cpv. D4cpv-Casq1 therefore provides substantial improvements in sensitivity, specificity, and reproducibility over existing monitors of SR free Ca(2+) concentration...
  10. pmc Paradoxical buffering of calcium by calsequestrin demonstrated for the calcium store of skeletal muscle
    Leandro Royer
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, Chicago, IL 60612, USA
    J Gen Physiol 136:325-38. 2010
    ..The study revealed a surprisingly modest loss of Ca(2+) storage capacity in null cells, which may reflect concurrent changes, rather than detract from the physiological importance of calsequestrin...
  11. pmc The cell boundary theorem: a simple law of the control of cytosolic calcium concentration
    Eduardo Rios
    Section of Cellular Signaling, Department of Molecular Biophysics and Physiology, Rush University, 1750 Chicago, IL 60612, USA
    J Physiol Sci 60:81-4. 2010
    ..While the law is discussed in terms of Ca(2+) homeostasis, it applies to any solute that may be transported by the plasma membrane...
  12. pmc Hyperactive intracellular calcium signaling associated with localized mitochondrial defects in skeletal muscle of an animal model of amyotrophic lateral sclerosis
    Jingsong Zhou
    Department of Molecular Biophysics and Physiology, Rush University Medical School, Chicago, Illinois 60612, USA
    J Biol Chem 285:705-12. 2010
    ..Our data reveal that mitochondria regulate Ca(2+) signaling in skeletal muscle, and loss of this capacity may contribute to the progression of muscle atrophy in ALS...
  13. pmc Voltage-gated proton channels maintain pH in human neutrophils during phagocytosis
    Deri Morgan
    Department of Molecular Biophysics and Physiology, Rush University Medical Center, Chicago, IL 60612, USA
    Proc Natl Acad Sci U S A 106:18022-7. 2009
    ..Both the rate and extent of acidification in HVCN1-deficient cells were twice larger than in control cells. In summary, acid extrusion by proton channels is essential to the production of reactive oxygen species during phagocytosis...
  14. pmc Deconstructing calsequestrin. Complex buffering in the calcium store of skeletal muscle
    Leandro Royer
    Department of Molecular Biophysics and Physiology, Rush University School of Medicine, Chicago, IL 60612, USA
    J Physiol 587:3101-11. 2009
    ..The experiments therefore suggest that other molecules are capable of providing sites for reversible binding of large amounts of Ca(2+) inside the sarcoplasmic reticulum...
  15. pmc Indo-1 derivatives for local calcium sensing
    Michael Bannwarth
    Institute of Chemical Sciences and Engineering, Ecole Polytechnique Federale de Lausanne EPFL, CH 1015, Lausanne, Switzerland
    ACS Chem Biol 4:179-190. 2009
    ....
  16. pmc Altered Ca2+ concentration, permeability and buffering in the myofibre Ca2+ store of a mouse model of malignant hyperthermia
    Carlo Manno
    S L Hamilton E RĂ­os Rush University School of Medicine, Department of Molecular Biophysics and Physiology, 1750 West Harrison St, Suite 1279JS, Chicago, IL 60612, USA Email
    J Physiol 591:4439-57. 2013
    ..The unstable SR buffering, mimicked by silencing of calsequestrin, may help precipitate the loss of Ca2+ control that defines a fulminant MH event...

Research Grants30

  1. FKBP Interaction with RYR Calcium Channels in Heart
    Donald M Bers; Fiscal Year: 2013
    ..We will conduct fundamental quantitative molecular fluorescence studies that will clarify both structurally and functionally how FKBP binds to and regulates RyR function in the heart. ..
  2. DEGENERATIVE AND DEMENTING DISEASES OF AGING
    Stanley B Prusiner; Fiscal Year: 2013
    ..The ultimate goal of all the proposed studies is to define the molecular events that feature in the formation of human prions in order to develop therapeutics that cure the human prion diseases. ..
  3. Roles of voltage sensor, S100A1 and calmodulin in skeletal muscle Ca2+ signaling
    Martin F Schneider; Fiscal Year: 2013
    ..Thus, this project has high impact for multiple disciplines, and for problems of both locomotion and breathing common to a variety of advanced diseased states and aging. ..
  4. Excitation-contraction Coupling in Normal and Dystrophic Mammalian Muscle
    Julio L Vergara; Fiscal Year: 2013
    ..The results will significantly broaden our understanding of muscle disease mechanisms and will potentially provide therapeutic molecular tools which are deemed necessary for further advances in gene therapy. ..
  5. Mechanism and function of a novel purinergic signaling cascade in skeletal muscle
    ANDREW ALVIN VOSS; Fiscal Year: 2013
    ....
  6. Molecular Analyses and Interventions for Biodefense and Emerging Pathogens
    Olaf Schneewind; Fiscal Year: 2013
    ..Research and training at the GLRCE is governed by a mechanism involving ongoing review of scientific excellence and translational goals, inter-institutional advisory boards and external scientific advisory bodies. ..
  7. Ca Signaling in Progression of Amyotrophic Lateral Sclerosis in Skeletal Muscle
    Jingsong Zhou; Fiscal Year: 2013
    ....
  8. Neurohumoral control of veins in hypertension
    Gregory D Fink; Fiscal Year: 2013
    ..This project tests the idea that altered structure or function of veins also may cause hypertension, and that it may be possible to treat hypertension using drugs that affect veins. ..
  9. Cellular Senescence and Aging
    James L Kirkland; Fiscal Year: 2013
    ..Our approach will provide timely, innovative, and clinically relevant interventional results based on addressing the fundamental question of the role of cellular senescence that has remained unanswered for many years. ..