It Is the central tenant of Core B and this program project that the most appropriate method to study MH is in murine models that express frequently observed human MH mutations and in genotyped human tissue. Mouse models provide sufficient tissues for molecular, biochemical, cellular, physiological and morphologic analyses by a multidisciplinary team whose collective goal is to understand this myopathy. Genotyped human myotubes will allow direct correlation of the observed phenotypes between humans and mice with the same mutations and fixed and frozen human tissues will allow morphologic and biochemical comparisons. Core B will perform repetitive tasks that are necessary to support all three Projects and Core D. This core will produce and genotype the 4 existing MH "knock-in" mice, 2 transgenic mice, and breed, genotype and maintain MH mice crossed with dnTrp6 and SERCA1 overexpression transgenic mice. It will also assure that the long-term pharmacological interventions with salicylamine and 4-OH-BDE49 are carried out. It will distribute these mice or frozen muscles for study by all three Projects as needed and provide fixed muscles from these mice at different ages and genders to Core D. Core B will make myoblast cell lines from all heterozygous and homozygous MH "knock-in" mice, to be used by all 3 projects. If needed homozygous myoblasts will be obtained from El8 embryos from timed matings in cases where there is homozygous lethality as was the case for the R163C and Y522S RyRI mice. Core B will receive, expand and maintain genotyped human MHS myoblasts from Core C and distribute these to the projects. It will also distribute genotyped glutaraldehyde fixed human biopsy samples to Core D and frozen muscle samples to Project 2 received from Core C. Core B has and will receive new MHS mutations from Core C Discovery, prepare mutated cDNA constructs in mammalian expression vectors for project 3, and where possible lentiviral vectors and permanently transduced cells and where not ,HSV vectors to be distributed to projects 1 and 2 for their analyses. The uniform and consistent supply of exactly the same study models to all three Projects and Core D by this Core will allow a truly integrated approach to the study of malignant hyperthermia.
Core B will distribute genotyped malignant hyperthermia susceptible (MHS) mice or mouse muscle, cross breed mice and administer pharmacologic agents, create and distribute MHS murine myoblast cell lines, expand human genotyped myoblast cells supplied by Core C and will act as the distribution point for snap frozen discarded MHS human biopsy tissue, glutaraldehyde fixed discarded MHS human biopsy tissue and human MHS myoblasts supplied by Core C to all three projects and Core D.
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|Burr, Adam R; Millay, Douglas P; Goonasekera, Sanjeewa A et al. (2014) Na+ dysregulation coupled with Ca2+ entry through NCX1 promotes muscular dystrophy in mice. Mol Cell Biol 34:1991-2002|
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|Boncompagni, Simona; Protasi, Feliciano; Franzini-Armstrong, Clara (2012) Sequential stages in the age-dependent gradual formation and accumulation of tubular aggregates in fast twitch muscle fibers: SERCA and calsequestrin involvement. Age (Dordr) 34:27-41|
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|Yuen, Benjamin; Boncompagni, Simona; Feng, Wei et al. (2012) Mice expressing T4826I-RYR1 are viable but exhibit sex- and genotype-dependent susceptibility to malignant hyperthermia and muscle damage. FASEB J 26:1311-22|
|Giulivi, Cecilia; Ross-Inta, Catherine; Omanska-Klusek, Alicja et al. (2011) Basal bioenergetic abnormalities in skeletal muscle from ryanodine receptor malignant hyperthermia-susceptible R163C knock-in mice. J Biol Chem 286:99-113|
|Feng, Wei; Barrientos, Genaro C; Cherednichenko, Gennady et al. (2011) Functional and biochemical properties of ryanodine receptor type 1 channels from heterozygous R163C malignant hyperthermia-susceptible mice. Mol Pharmacol 79:420-31|
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