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DP71 和 DP40 同工型在增殖和分化神经干细胞中的差异表达:DP40 剪接变异体的鉴定。

Differential expression of Dp71 and Dp40 isoforms in proliferating and differentiated neural stem cells: Identification of Dp40 splicing variants.

机构信息

Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV), Av. Instituto Politécnico Nacional 2508, Col. San Pedro Zacatenco, 07360, Ciudad de México, Mexico.

Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV), Av. Instituto Politécnico Nacional 2508, Col. San Pedro Zacatenco, 07360, Ciudad de México, Mexico.

出版信息

Biochem Biophys Res Commun. 2021 Jun 30;560:152-158. doi: 10.1016/j.bbrc.2021.03.142. Epub 2021 May 12.

Abstract

Dp71 and Dp40 are the main products of the DMD gene in the central nervous system, and they are developmentally regulated from the early stages of embryonic development to adulthood. To further study the roles of Dp71 and Dp40 during cell proliferation and neural differentiation, we analyzed Dp71/Dp40 isoform expression at the mRNA level by RT-PCR assays to identify alternative splicing (AS) in the isoforms expressed in rat neural stem/progenitor cells (NSPCs) and in differentiated cells (neurons and glia). We found that proliferating NSPCs expressed Dp71d, Dp71d, Dp71f, Dp71f, Dp71d and Dp40, as well as two Dp40 isoforms: Dp40 and Dp40. In differentiated cells we also found the expression of Dp71d, Dp71d, Dp71f, Dp71f and Dp40. However, the expression frequencies were different in both stages. In addition, in differentiated cells, we found Dp71f, and interestingly, we did not find the expression of Dp71d or the newly identified Dp40 isoforms. In this work we show that NSPC differentiation is accompanied by changes in Dp71/Dp40 isoform expression, suggesting different roles for these isoforms in NSPCs proliferation and neuronal differentiation, and we describe, for the first time, alternative splicing of Dp40.

摘要

Dp71 和 Dp40 是 DMD 基因在中枢神经系统中的主要产物,它们从胚胎发育的早期到成年期都受到发育调控。为了进一步研究 Dp71 和 Dp40 在细胞增殖和神经分化过程中的作用,我们通过 RT-PCR 分析了 Dp71/Dp40 异构体在大鼠神经干细胞/祖细胞 (NSPCs) 和分化细胞 (神经元和神经胶质) 中的 mRNA 水平表达,以鉴定异构体中的选择性剪接 (AS)。我们发现增殖的 NSPCs 表达 Dp71d、Dp71d、Dp71f、Dp71f、Dp71d 和 Dp40,以及两种 Dp40 异构体:Dp40 和 Dp40。在分化细胞中,我们还发现了 Dp71d、Dp71d、Dp71f、Dp71f 和 Dp40 的表达。然而,在这两个阶段的表达频率不同。此外,在分化细胞中,我们发现了 Dp71f,有趣的是,我们没有发现 Dp71d 或新鉴定的 Dp40 异构体的表达。在这项工作中,我们表明 NSPC 分化伴随着 Dp71/Dp40 异构体表达的变化,表明这些异构体在 NSPCs 增殖和神经元分化中具有不同的作用,并且我们首次描述了 Dp40 的选择性剪接。

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