Murine Reagents

它是核心B和这个项目的中心租户，研究MH最合适的方法是在经常观察到的人类MH突变的小鼠模型中和在人类组织中进行基因分型。 小鼠模型为多学科小组进行分子、生化、细胞、生理和形态分析提供了足够的组织，他们的共同目标是了解这种肌病。基因分型的人类肌管将允许观察到的人类和小鼠之间的表型直接相关 突变和固定和冷冻的人体组织将可以进行形态和生化比较。 核心B将执行支持所有三个项目和核心D所需的重复性任务。该核心将生产现有的4只MH“敲入”小鼠和2只转基因小鼠并对其进行基因分型，并培育、分型和维持与dnTrp6和SERCA1高表达转基因小鼠杂交的MH小鼠。它还将确保对水杨胺和4-羟基-BDE49进行长期药理干预。会的 根据需要分配这些小鼠或冰冻肌肉供所有三个项目研究，并将这些小鼠不同年龄和性别的固定肌肉提供给Core D。 核心B将从所有杂合子和纯合子的MH“敲入”小鼠中制造成肌细胞系，供所有三个项目使用。如果需要，在存在纯合子致死性的情况下，可以从定时配对的El8胚胎中获得纯合子成肌细胞，就像R163C和Y522S RyRI小鼠一样。 核心B将接收、扩大和维护来自核心C的基因分型的人类MHS成肌细胞，并将这些细胞分发给项目。它还将向核心D分发戊二醛固定的人体活检样本，并将冷冻肌肉样本分发给从核心C接收的项目2。 核心B已经并将从核心C发现获得新的MHS突变，在项目3的哺乳动物表达载体中准备突变的cDNA结构，在可能的情况下准备慢病毒载体和永久转导细胞，如果没有，则将HSV载体分发到项目1和项目2进行分析。 这一核心公司向所有三个项目和核心D提供完全相同的研究模型的统一和一致的方法将使恶性热疗的研究真正整合起来。

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.