Analysis and Therapy of Muscular Dystrophy in Zebrafish

在过去的5年里，我们一直致力于将斑马鱼建立为肌肉萎缩的模型。 营养不良在这方面，我们已经发表了斑马鱼缺乏抗肌萎缩蛋白和8- 肌聚糖，完成了一个大的遗传筛选，以分离额外的营养不良突变体，并确定了 突变体基因中的一种。利用我们在肌肉研究和建立 斑马鱼作为疾病模型，我们现在建议使用鱼类来研究肌肉萎缩症的发病机制。 营养不良和评估细胞疗法作为一种潜在的治疗选择。 在这个项目中的第一个目标是充分描述我们现有的两个营养不良的斑马鱼 模型（EMZ和SOF），目的是更好地了解肌营养不良症的发病机制。这些 突变体显示出与肌营养不良蛋白突变体（SAPJE）非常相似的肌肉变性表型， 这种表型是肌营养不良症的症状我们建议确定基因突变， 这些突变体使用传统的作图方法，然后测序候选基因，以确定 特殊突变如果这些人类基因目前与肌肉萎缩症无关， 这些基因将被认为是疾病候选基因，并在人类患者中测序， 肌肉萎缩症的病因不明。由于看似无关的蛋白质突变可以表现为 因此，在肌营养不良症中，鉴定额外的基因将有助于建立疾病途径。 其次，我们已经建立了在斑马鱼体内移植细胞群的方法， 发展阶段，现在建议使用这个系统来识别最有能力的细胞群， 植入并矫正患病肌肉。肌肉移植细胞的基因表达谱 将群体与非移植细胞进行比较，以鉴定主要在移植细胞中表达的基因。 移植细胞差异表达的基因将被认为是潜在的标记，并用于纯化 用于未来的实验和治疗。最后，我们计划解剖 通过分析斑马鱼的发育潜力来研究不同干细胞群体的谱系关系 肌肉祖细胞这将通过早期移植有限的标记细胞群来实现 然后随着鱼的成熟而改变命运

Over the last 5 years, we have been working to establish the zebrafish as a model for muscular dystrophy. In this capacity, we have published the phenotype of zebrafish lacking dystrophin and 8- sarcoglycan, completed a large genetic screen to isolate additional dystrophic mutants, and identified the mutant gene in the runzelmutant. Using our experiences in muscle research and in establishing the zebrafish as a disease model, we now propose to use the fish to investigate the pathogenesis of muscular dystrophy and evaluate cell therapy as a potential treatment option. The first aim in this project proposes to fully characterize two of our available dystrophic zebrafish models (emz and sof) with the goal of better understanding the pathogenesis of muscular dystrophy. These mutants show a muscle degeneration phenotype very similar to the dystrophin mutant (sapje) suggesting that this phenotype is symptomatic of muscular dystrophy. We propose to identify the genetic mutations in these mutants using a traditional mapping approach and then sequencing candidate genes to identify the specific mutation. If the orthologous human genes are not currently associated with muscular dystrophy, these genes will be considered disease candidate genes and sequenced in human patients for which the cause of muscular dystrophy is unknown. Since mutations in seemingly unrelated proteins can manifest as muscular dystrophy, the identification of additional genes would be helpful for establishing disease pathways. Secondly, we have established methods to transplant cell populations in zebrafish at all developmental stages and now propose using this system to identify the cell population most capable of engrafting into and correcting the diseased muscle. Gene expression profiles of muscle engrafting cell populations will be compared with non-engrafting cells to identify genes expressed predominantly in the engrafting cells. Differentially expressed genes will be considered potential markers and used to purify analogous cell populations in mammals for future experimentation and therapy. Finally, we plan to dissect the lineage relationship of various stem cell populations by assaying the developmental potential of zebrafish muscle progenitor cells. This will be accomplished by transplanting limited populations of labeled cells early in development and then following their fate as the fish matures.