Murine Reagents

它是核心B的核心租户，并且该项目项目认为，研究MH的最合适方法是在表达经常观察到的人MH突变的鼠模型和基因分型的人组织中。 小鼠模型提供了足够的组织分子，生物化学，细胞，生理和形态学分析的多学科小组的集体目标是了解这种肌病。基因分型的人肌管将允许人类和小鼠之间观察到的表型的直接相关性， 突变和固定和冷冻的人体组织将允许形态学和生物化学比较。 核心B将执行支持所有三个项目和核心D所必需的重复性任务。该核心将产生4只现有MH“敲入”小鼠和2只转基因小鼠并对其进行基因分型，并繁殖、基因分型和维持与dnTrp 6和SERCA 1过表达转基因小鼠杂交的MH小鼠。它还将确保使用水杨胺和4-OH-BDE 49进行长期药理干预。它将 根据需要将这些小鼠或冷冻肌肉分配给所有三个项目进行研究，并将不同年龄和性别的这些小鼠的固定肌肉提供给核心D。 核心B将从所有杂合和纯合MH“敲入”小鼠中制备成肌细胞系，供所有3个项目使用。如果需要，在存在纯合致死性的情况下，如R163 C和Y 522 S RyRI小鼠的情况，将从来自定时交配的E18胚胎获得纯合成肌细胞。 核心B将接收、扩增和维持来自核心C的基因分型人MHS成肌细胞，并将其分发给项目。其还将向核心D分发基因型戊二醛固定的人体活检样本，并向项目2分发从核心C收到的冷冻肌肉样本。 核心B已经并将从核心C Discovery获得新的MHS突变，在项目3的哺乳动物表达载体中制备突变的cDNA构建体，以及在可能的情况下，将慢病毒载体和永久转导的细胞以及HSV载体分配给项目1和2进行分析。 该核心向所有三个项目和核心D统一和一致地提供完全相同的研究模型，将使恶性高热的研究成为一种真正的综合方法。

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.