Division of Genetics, Program in Genomics, Children's Hospital, Boston, MA 02115, USA.
Proc Natl Acad Sci U S A. 2011 Mar 29;108(13):5331-6. doi: 10.1073/pnas.1102116108. Epub 2011 Mar 14.
Two known zebrafish dystrophin mutants, sapje and sapje-like (sap(c/100)), represent excellent small-animal models of human muscular dystrophy. Using these dystrophin-null zebrafish, we have screened the Prestwick chemical library for small molecules that modulate the muscle phenotype in these fish. With a quick and easy birefringence assay, we have identified seven small molecules that influence muscle pathology in dystrophin-null zebrafish without restoration of dystrophin expression. Three of seven candidate chemicals restored normal birefringence and increased survival of dystrophin-null fish. One chemical, aminophylline, which is known to be a nonselective phosphodiesterase (PDE) inhibitor, had the greatest ability to restore normal muscle structure and up-regulate the cAMP-dependent PKA pathway in treated dystrophin-deficient fish. Moreover, other PDE inhibitors also reduced the percentage of affected sapje fish. The identification of compounds, especially PDE inhibitors, that moderate the muscle phenotype in these dystrophin-null zebrafish validates the screening protocol described here and may lead to candidate molecules to be used as therapeutic interventions in human muscular dystrophy.
两种已知的斑马鱼肌营养不良蛋白突变体，sapje 和 sapje-like（sap(c/100)），代表了人类肌肉营养不良症的优秀小型动物模型。我们使用这些肌营养不良蛋白缺失的斑马鱼，在 Prestwick 化学文库中筛选了能够调节这些鱼类肌肉表型的小分子。通过快速简便的双折射测定法，我们已经鉴定出七种小分子，它们可以影响肌营养不良蛋白缺失的斑马鱼的肌肉病理学，而不恢复肌营养不良蛋白的表达。在七种候选化学物质中，有三种能够恢复正常的双折射并提高肌营养不良蛋白缺失鱼类的存活率。一种化学物质氨茶碱，已知是一种非选择性磷酸二酯酶（PDE）抑制剂，它具有最大的能力来恢复正常的肌肉结构，并上调治疗后肌营养不良蛋白缺陷鱼类中的 cAMP 依赖性 PKA 途径。此外，其他 PDE 抑制剂也降低了受影响的 sapje 鱼的百分比。在这些肌营养不良蛋白缺失的斑马鱼中，能够调节肌肉表型的化合物的鉴定，特别是 PDE 抑制剂，验证了这里描述的筛选方案，并可能导致候选分子被用作人类肌肉营养不良症的治疗干预措施。
