Aleman V, Osorio B, Chavez O, Rendon A, Mornet D, Martinez D
Department of Fisiologia, Biofisica y Neurociencias, Cinvestav, IPN. Apdo. Postal 14-740, Mexico, DF, CP 07000, Mexico.
Histochem Cell Biol. 2001 Mar;115(3):243-54. doi: 10.1007/s004180000221.
It has been suggested that the absence or altered structure of Dp71, a C-terminal dystrophin gene encoded protein, is responsible for mental alterations observed in about 30% of Duchenne muscular dystrophy patients. Most of these patients have premature translational termination or point mutations at the C-terminal domain of this gene. In brain, Dp71 is the major protein product of the dystrophin gene. To determine the function of Dp71 isoforms in this organ, it is important to document their presence and intracellular localization in brain cells. Extracts from cultured hippocampal neurons and forebrain astrocytes and 5F3 and Dys 2 monoclonal antibodies were thus used for western blots. In these conditions, two Dp71 isoforms spliced or not at exon 78 were detected in both cells (Dp71f and Dp71d, respectively). By immunocytochemistry, we mapped Dp71f and Dp71d in the Golgi complex (GC) and in neuronal nuclei. Only Dp71d was found in cytoplasmic neurofilaments. In astrocytes, these isoforms were detected in the GC. These cell localization data suggest that these Dp71 isoforms may have different functions in the same cell or organelle, as well as in the two different cells analyzed.
有人提出，Dp71（一种由肌营养不良蛋白基因C端编码的蛋白质）的缺失或结构改变，是导致约30%的杜氏肌营养不良患者出现精神改变的原因。这些患者中的大多数在该基因的C端结构域存在过早的翻译终止或点突变。在大脑中，Dp71是肌营养不良蛋白基因的主要蛋白质产物。为了确定Dp71亚型在该器官中的功能，记录它们在脑细胞中的存在情况和细胞内定位非常重要。因此，使用来自培养的海马神经元和前脑星形胶质细胞的提取物以及5F3和Dys 2单克隆抗体进行蛋白质印迹分析。在这些条件下，在两种细胞中均检测到了在第78外显子处剪接或未剪接的两种Dp71亚型（分别为Dp71f和Dp71d）。通过免疫细胞化学，我们将Dp71f和Dp71d定位在高尔基体复合体（GC）和神经元细胞核中。仅在细胞质神经丝中发现了Dp71d。在星形胶质细胞中，在GC中检测到了这些亚型。这些细胞定位数据表明，这些Dp71亚型在同一细胞或细胞器以及所分析的两种不同细胞中可能具有不同的功能。
