Establishment of stable zebrafish genetic models for studying myotonic dystrophy

项目总结/摘要 强直性肌营养不良症（DM）是肌营养不良症的最常见形式之一， 每八千人中就有一人感染糖尿病患者常见消化道症状， 涉及改变肠道运动，包括胃排空减慢， 运动、腹痛、腹泻和便秘。肠道细菌发挥重要作用 在消化中，许多DM患者有这些细菌的过度生长。四分之一DM 患者将消化道症状描述为疾病的最大致残方面，但这些 症状知之甚少。这项拟议的工作使用斑马鱼来研究DM，因为 消化症状可以直接在活的透明幼虫中观察到， 产生大量的后代，可用于筛选药物，以改善糖尿病 症状 DM最常见的形式是由含有长重复序列的RNA表达引起的 CUG核苷酸，其结合并隔离MBNL蛋白质家族，并防止它们 调节一个重要的分子过程称为选择性剪接。斑马鱼已经被 - 产生通过mbnl基因突变模拟DM中MBNL蛋白功能丧失的蛋白，和 这些鱼显示出一些与人类疾病中相同的选择性剪接变化。 通过表达CUG重复RNA，鱼也被制造成以另一种方式模拟DM。 使用新的DM模式鱼，第一个目标是确定它们是否具有DM样 症状，并确定有助于这些症状的分子变化。下一个目标 是使用显微镜测量活体斑马鱼DM模型中肠道运动的变化， 确定细菌是否有助于这些变化，并确定哪些组织类型 对这些症状很重要。最后，对候选药物进行小规模试验， 将进行模型鱼的DM样症状。这将作为以下方面的原理证明： 未来大规模筛选改善鱼类DM样症状的药物， 目的是确定减少或消除糖尿病患者症状的药物。 总的来说，这些研究将提供重要的洞察力，了解鲜为人知的消化道 糖尿病的症状及其背后的分子机制。他们还将创造新的 动物模型工具，这将是非常宝贵的研究其他方面的糖尿病病理学和 筛选药物以改善糖尿病患者的生活。 !

Project Summary/Abstract Myotonic dystrophy (DM) is one of the most common forms of muscular dystrophy, affecting 1 in 8,000 people. It is common for DM patients to have digestive symptoms, often involving altered gut movement, including slowed stomach emptying, slowed intestinal movement, abdominal pain, diarrhea, and constipation. Intestinal bacteria play important roles in digestion, and many DM patients have overgrowth of these bacteria. A quarter of DM patients describe digestive symptoms as the most disabling aspect of the disease, yet these symptoms are poorly understood. The proposed work uses zebrafish to study DM because digestive symptoms can be observed directly in live, transparent larvae, and because zebrafish produce large numbers of offspring that can be used to screen for drugs to improve DM symptoms. The most common form of DM is caused by expression of RNA containing long repeats of CUG nucleotides, which bind to and sequester the MBNL family of proteins and prevent them from regulating an important molecular process called alternative splicing. Zebrafish have been generated that mimic the loss of MBNL protein function in DM by mutation of mbnl genes, and these fish show some of the same alternative splicing changes that are seen in human disease. Fish are also being made that mimic DM in another way, by expressing CUG repeat RNA. Using the new DM model fish, the first goal is to determine whether they have DM-like symptoms and to identify molecular changes that contribute to these symptoms. The next goal is to measure changes in gut movement in live zebrafish DM models using microscopy, to determine whether bacteria contribute to these changes, and to determine which tissue types are important for these symptoms. Finally, a small-scale test of candidate drugs to ameliorate the DM-like symptoms of model fish will be performed. This will serve as a proof-of-principle for future large-scale screening for drugs that improve DM-like symptoms in fish with the long term goal of identifying drugs to reduce or eliminate symptoms in DM patients. Overall, these studies will provide important insight into the poorly understood digestive symptoms of DM and the molecular mechanisms behind them. They will also generate novel animal model tools that will be invaluable for studying other aspects of DM pathology and for screening for drugs to improve the lives of individuals with DM. !