Identification of Dp71 Isoforms Expressed in PC12 Cells: Subcellular Localization and Colocalization with β-Dystroglycan and α1-Syntrophin
Identification of Dp71 Isoforms Expressed in PC12 Cells: Subcellular Localization and Colocalization with β-Dystroglycan and α1-Syntrophin
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DOI：
10.1007/s12031-015-0657-8
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发表时间：
2016-02-01
影响因子：
3.1
通讯作者：
Montanez, Cecilia
中科院分区：
文献类型：
--
作者：
Aragon, Jorge;Martinez-Herrera, Alejandro;Montanez, Cecilia
已经描述了几种抗肌萎缩蛋白Dp 71信使RNA（mRNA）选择性剪接变体。根据外显子78或内含子77的剪接，Dp 71蛋白被分为Dp 71 d、Dp 71 f和Dp 71 e，并且每组都具有特定的C末端。在这项研究中，我们探讨了Dp 71异构体在互补DNA（cDNA）水平的表达和亚细胞定位的重组Myc-Dp 71蛋白在PC 12细胞。我们确定PC 12细胞表达Dp 71 a、Dp 71 c、Dp 71 ab、Dp 71 e和Dp 71 ec mRNA剪接变体。在未分化和神经生长因子分化的PC 12 Tet-ON细胞中，发现Dp 71 a、Dp 71 ab和Dp 71 e与β-肌营养不良蛋白聚糖和α 1-突触营养蛋白在外周/细胞质中定位和共定位，而Dp 71 c和Dp 71 ec主要定位在细胞外周，并且与β-肌营养不良蛋白聚糖和α 1-突触营养蛋白的共定位较少。与未分化的细胞相比，分化的PC 12 Tet-ON细胞的细胞核中Dp 71 a、Dp 71 e和Dp 71 ec的水平增加。Dp 71亚型也定位于神经突延伸和生长锥。
Several dystrophin Dp71 messenger RNA (mRNA) alternative splice variants have been described. According to the splicing of exon 78 or intron 77, Dp71 proteins are grouped as Dp71d, Dp71f, and Dp71e, and each group has a specific C-terminal end. In this study, we explored the expression of Dp71 isoforms at the complementary DNA (cDNA) level and the subcellular localization of recombinant Myc-Dp71 proteins in PC12 cells. We determined that PC12 cells express Dp71a, Dp71c, Dp71ab, Dp71e, and Dp71ec mRNA splice variants. In undifferentiated and nerve growth factor-differentiated PC12 Tet-ON cells, Dp71a, Dp71ab, and Dp71e were found to localize and colocalize with beta-dystroglycan and alpha 1-syntrophin in the periphery/cytoplasm, while Dp71c and Dp71ec were mainly localized in the cell periphery and showed less colocalization with beta-dystroglycan and alpha 1-syntrophin. The levels of Dp71a, Dp71e, and Dp71ec were increased in the nucleus of differentiated PC12 Tet-ON cells compared to undifferentiated cells. Dp71 isoforms were also localized in neurite extensions and growth cones.