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在PC12细胞中表达的Dp71亚型的鉴定:亚细胞定位以及与β-肌营养不良聚糖和α1-肌营养不良蛋白聚糖的共定位

Identification of Dp71 Isoforms Expressed in PC12 Cells: Subcellular Localization and Colocalization with β-Dystroglycan and α1-Syntrophin.

作者信息

Aragón Jorge, Martínez-Herrera Alejandro, Romo-Yáñez José, Ceja Víctor, Azotla-Vilchis Coztli, Siqueiros-Márquez Lourdes, Soid-Raggi Gabriela, Herrera-Salazar Alma, Montañez Cecilia

机构信息

Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Av. IPN No. 2508, San Pedro Zacatenco, C.P. 07360, México, D. F., Mexico.

Departamento de Genómica Humana, Instituto Nacional de Perinatología, C.P. 11000, México, D.F., Mexico.

出版信息

J Mol Neurosci. 2016 Feb;58(2):201-9. doi: 10.1007/s12031-015-0657-8. Epub 2015 Sep 28.

Abstract

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 β-dystroglycan and α1-syntrophin in the periphery/cytoplasm, while Dp71c and Dp71ec were mainly localized in the cell periphery and showed less colocalization with β-dystroglycan and α1-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.

摘要

已经描述了几种肌营养不良蛋白Dp71信使核糖核酸(mRNA)可变剪接变体。根据外显子78或内含子77的剪接情况,Dp71蛋白可分为Dp71d、Dp71f和Dp71e,每组都有一个特定的C末端。在本研究中,我们在互补脱氧核糖核酸(cDNA)水平上探索了Dp71亚型的表达以及重组Myc-Dp71蛋白在PC12细胞中的亚细胞定位。我们确定PC12细胞表达Dp71a、Dp71c、Dp71ab、Dp71e和Dp71ec mRNA剪接变体。在未分化和神经生长因子分化的PC12 Tet-ON细胞中,发现Dp71a、Dp71ab和Dp71e在外周/细胞质中与β-肌营养不良聚糖和α1-肌营养不良蛋白共定位,而Dp71c和Dp71ec主要定位于细胞外周,与β-肌营养不良聚糖和α1-肌营养不良蛋白的共定位较少。与未分化细胞相比,分化的PC12 Tet-ON细胞细胞核中Dp71a、Dp71e和Dp71ec的水平升高。Dp71亚型也定位于神经突延伸和生长锥中。

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