Generation and characterization of a Cre-Lox regulated transgenic zebrafish model of SBMA

脊髓性和延髓性肌萎缩症(SBMA或肯尼迪病)是一种缓慢进展的X连锁疾病 影响男性的神经肌肉疾病。它是由雄激素内CAG重复序列的扩张引起的。 受体(AR)基因，编码蛋白质中的谷氨酰胺链。在没有治愈或治疗的情况下，发现潜力 针对这一患者群体的治疗干预迫在眉睫。我们的目标是创建一个CRE诱导型、低成本和 使用斑马鱼的SBMA高通量模型。斑马鱼研究的发现是非常重要的 可翻译给人类，因为斑马鱼是脊椎动物，与人类具有广泛的结构同源性 神经解剖学和肌肉生理学。更具体地说，对于SBMA患者，斑马鱼表达的AR是 在结构和功能上与人类AR高度同源。人AR转基因的Cre诱导性将使 生产能够表达高水平有毒的多谷氨酰胺扩展AR的方正菌株。我们 假设这将导致斑马鱼幼体的强烈运动功能障碍。这些斑马鱼幼体模拟SBMA 将服从于快速(超过1-2周)化学和药物库筛选，或基因修改(S)，以 确定运动功能的调节器。这样的分析可以在几天内以非常低的成本建立起来 与啮齿动物模型中的此类检测相比。此外，组织特异性Cre斑马鱼品系可以与 这些SBMA斑马鱼在未来的研究中将探讨运动神经元相对于骨骼肌的相对作用 在SBMA发病机制中的作用。更好地理解其中每一个的相对作用 疾病中的细胞类型将支持SBMA患者的临床试验设计。此外，因为SBMA共享财产 对于其他神经退行性疾病、神经肌肉疾病和多谷氨酰胺疾病，在 SBMA斑马鱼模型可能也适用于其中一些其他疾病。

Spinal and bulbar muscular atrophy (SBMA or Kennedy's disease) is a slowly progressive, X-linked neuromuscular disease affecting men. It is caused by the expansion of a CAG repeat within the androgen receptor (AR) gene, encoding a glutamine tract in the protein. With no cure or therapy, identifying potential therapeutic interventions for this patient population is pressing. We aim to create a Cre-inducible, low cost, and high-throughput model of SBMA using zebrafish. Discoveries made from zebrafish research are highly translatable to humans since zebrafish are vertebrates with extensive structural homology to human neuroanatomy and muscle physiology. More specifically for SBMA patients, zebrafish express an AR that is highly homologous in structure and function to human AR. Cre-inducibility of the human AR transgene will enable the production of founder lines that are able to express high levels of the toxic polyglutamine-expanded AR. We hypothesize this will lead to robust motor dysfunction in larval zebrafish. These larval zebrafish modeling SBMA will be amenable to quick (over 1-2 weeks) chemical and drug library screens, or genetic modification(s) to identify modulators of motor function. Such assays could be set up within only a few days and at a very low cost compared to such assays in rodent models. Moreover, tissue specific Cre zebrafish lines can be crossed with these SBMA zebrafish in future studies to investigate the relative role of motor neurons verses skeletal muscle in the onset and progression of SBMA pathogenesis. A better understanding of the relative roles of each of these cell types in disease will support clinical trial design for SBMA patients. Further, because SBMA shares properties with other neurodegenerative, neuromuscular, and polyglutamine diseases, therapeutic targets identified in the SBMA zebrafish model may also be applicable to some of these other disorders.