The long-term objective in Project 2 is to gain understanding of the biochemical and biophysical mechanisms by which mutations from each MH/CCD hot spot alter key regulatory functions of the RyR1 Ca2+ channel complex, and how these changes deregulate SR Ca2+ transport in skeletal muscle SR, and dendritic cells that express MH/CCD RyR1. HYPOTHESIS I) MH/CCD mutations alter cytoplasmic and luminal regulation of RyR1 by ligands by changing the activation energy (Ea) needed for conformational transitions of the channel. A1.1. Define differences among 11 MH/CCD and wild type (Wt) genotypes on expression of key triadic proteins, SR loading capacity and their relationship to altered cation interactions at H- and Lsites using equilibrium [3H]ryanodine ([3H]Ry) binding analysis. A1.2. To analyze mechanisms responsible for altered regulation by cytoplasmic Ca2+, Mg2+, and ATP and luminal Ca2+ anc calsequestrin (CSQ) using selected RyR1 channels. A1.3. Compare the differences in activation energy (Ea) for selected MH/CCD mutations. A1.4. Analyze how halothane and dantrolene differentially influence the stability of closed and open states of selected MH/CCD RyR1s. HYPOTHESIS II) MH/CCD mutations disrupt the redox-sensing properties of RyR1. A2.1. To establish if differences in transmembrane redox regulation and GSH/GSSG transport contribute to the pathophysiology of MH/CCD. HYPOTHESIS III) MH/CCD mutations produce a dysfunctional phenotype in dendritic cells. A3.1. Elucidate how MH/CCD mutations alter murine dendritic cell activation, dendritic cell secretion of IL-6 mediated by ATP acting at P2Y receptors, and if and how MH/CCD mutations influence dendritic cell activation of T-cells. A3.2. To extend these studies to DCs cultured from human blood.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Program Projects (P01)
Project #
5P01AR052354-05
Application #
8054283
Study Section
Special Emphasis Panel (ZAR1)
Project Start
Project End
Budget Start
2010-04-01
Budget End
2011-03-31
Support Year
5
Fiscal Year
2010
Total Cost
$270,659
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Lavorato, Manuela; Loro, Emanuele; Debattisti, Valentina et al. (2018) Elongated mitochondrial constrictions and fission in muscle fatigue. J Cell Sci 131:
Glaser, Nosta; Iyer, Ramesh; Gilly, William et al. (2018) Functionally Driven Modulation of Sarcomeric Structure and Membrane Systems in the Fast Muscles of a Copepod (Gaussia princeps). Anat Rec (Hoboken) 301:2164-2176
Polster, Alexander; Nelson, Benjamin R; Papadopoulos, Symeon et al. (2018) Stac proteins associate with the critical domain for excitation-contraction coupling in the II-III loop of CaV1.1. J Gen Physiol 150:613-624
Riazi, Sheila; Kraeva, Natalia; Hopkins, Philip M (2018) Malignant Hyperthermia in the Post-Genomics Era: New Perspectives on an Old Concept. Anesthesiology 128:168-180
Zheng, Jing; Chen, Juan; Zou, Xiaohan et al. (2018) Saikosaponin d causes apoptotic death of cultured neocortical neurons by increasing membrane permeability and elevating intracellular Ca2+ concentration. Neurotoxicology 70:112-121
Holland, Erika B; Goldstone, Jared V; Pessah, Isaac N et al. (2017) Ryanodine receptor and FK506 binding protein 1 in the Atlantic killifish (Fundulus heteroclitus): A phylogenetic and population-based comparison. Aquat Toxicol 192:105-115
Perni, Stefano; Lavorato, Manuela; Beam, Kurt G (2017) De novo reconstitution reveals the proteins required for skeletal muscle voltage-induced Ca2+ release. Proc Natl Acad Sci U S A 114:13822-13827
Lavorato, Manuela; Iyer, V Ramesh; Dewight, Williams et al. (2017) Increased mitochondrial nanotunneling activity, induced by calcium imbalance, affects intermitochondrial matrix exchanges. Proc Natl Acad Sci U S A 114:E849-E858
Zhang, Rui; Pessah, Isaac N (2017) Divergent Mechanisms Leading to Signaling Dysfunction in Embryonic Muscle by Bisphenol A and Tetrabromobisphenol A. Mol Pharmacol 91:428-436
Linsley, Jeremy W; Hsu, I-Uen; Groom, Linda et al. (2017) Congenital myopathy results from misregulation of a muscle Ca2+ channel by mutant Stac3. Proc Natl Acad Sci U S A 114:E228-E236

Showing the most recent 10 out of 78 publications