A study of pathogenesis for Duchenne muscular dystrophy

(1)抗肌萎缩蛋白和抗肌萎缩蛋白相关蛋白：为了确定抗肌萎缩蛋白的定位和功能意义，我们研究了几乎整个身体的各种组织的控制和mdx小鼠，与免疫组化，使用8种抗肌萎缩蛋白的单克隆和多克隆抗体。我们证实肌营养不良蛋白确实存在于神经肌肉接头、肌腱接头和梭内肌纤维等神经元和突触上。用RT-PCR方法在视网膜上发现了dystrophin及其四种亚型。肌营养不良蛋白在中枢神经系统和突触中的定位表明其在传导系统中的生理功能而不是机械功能。肌营养不良蛋白相关蛋白（DRP）也存在于与肌营养不良蛋白剂量相同的区域。在mdx组织中，突触上的DRP增加似乎是对dystrophin缺失的代偿反应。这一结果也提示Dystrophin和DRP在传导系统中具有相同的生理功能。(2)神经生长因子受体：用神经生长因子受体（NGFR）抗体对肌营养不良患者的肌肉组织进行免疫组化和免疫组织化学研究。结果显示，肌营养不良患者活检肌肉标本中血管外膜图尼卡上NGFR免疫反应性较强，但非诊断对照标本中几乎不存在。这表明在交感神经系统中的一个过程，涉及肌肉营养不良症的血管。(3)点突变和携带者的检测：我们证明了TOP分析在DMD患者和携带者女性中检测来自血液样品的材料的无义突变的有效性。

(1) Dystrophin and Dystophin-related protein: To determine the localization and functional significance of dystrophin, we studied various tissues from almost the entire body of control and mdx mice, with immunohistochemically, immunoelectrophoretically, using eight kinds of monoclonal and polyclonal antibodies against dystrophin. We confirmed that dystrophin is really exist on neurons and synapses such as neuromuscular junctions, myotendinous junctions, and intrafusal muscle fibers. We also found dystrophin and its four isoforms on retina with RT-PCR method. The localization of dystrophin in central nervous system and synapses suggests its physiological function in the conduction system rather than a mechanical one. Dystrophin-related protein (DRP) is also exist on the same regions as dystrophin dose. DRP seems to increase on synapses in mdx tissue as compensatory reaction for absent of dystrophin. The result also suggest that dystrophin and DRP have the same physiological function in the conduction system.(2) Nerve growth factor receptor: Immunohistochemical and immunoelectrophoretical studies were performed using nerve growth factor receptor (NGFR) antibody for muscle tissues from patients with muscular dystrophies. The results showed strong NGFR immunoreactivity on the tunica adventitia of blood vessels in biopsied muscle specimens from muscular dystrophy patients, but it was almost absent in specimens from non-diagnostic controls. This suggests a process in the sympathetic nervous system involving blood vessels in muscular dystrophies.(3) Detection of point mutation and carrier: We demonstrated its effectiveness of TOP-analysis to detect nonsense mutations in both DMD-patients and carrier females on material derived from blood samples.