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首页> 外文期刊>Scientific reports. >Regulation of Shootin1 Gene Expression Involves NGF-induced Alternative Splicing during Neuronal Differentiation of PC12 Cells
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Regulation of Shootin1 Gene Expression Involves NGF-induced Alternative Splicing during Neuronal Differentiation of PC12 Cells

机译：Sheain1基因表达的调节涉及在PC12细胞的神经元分化期间的NGF诱导的替代剪接

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Shootin1 is a protein involved in neuronal polarization, and has been shown to be a key molecule for the positiveegative feedback loop for axon induction required during neuronal symmetry breaking. To better understand the molecular basis of shootin1 dynamics, we analysed the regulatory pathways and the expressional status of shootin1 gene during NGF-induced neuronal differentiation. We demonstrated that the isoform-1 and isoform-2 of shootin1 is differentially expressed during neuronal differentiation. By blocking individual downstream pathways of NGF signalling, we found that PI3K/Akt pathway plays a major role in the expression of shootin1 isoform-2. Western blot and RT-PCR results showed that the isoform-1 of shootin1 is constitutively expressed, while the isoform-2 is expressed in a manner that is strictly dependent on NGF-stimulation. Isoform-specific RT-PCR results demonstrated that the differential expression of the isoform-1 and isoform-2 of shootin1 is a consequence of alternative splicing of shootin1 pre-mRNA, in response to NGF-signalling. Collectively these findings provide the first information on the molecular mechanisms regulating the expression of shootin1 gene and represent the first example of NGF-induced alternative splicing process that has a regulatory role in neuritogenesis.
机译：ShootIn1是参与神经元极化的蛋白质，并且已被证明是神经元对称性断裂所需的正/负反馈回路的关键分子。为了更好地理解ShootIn1动态的分子基础，我们分析了NGF诱导神经元分化期间SAMM1基因的调节途径和表达状态。我们证明，在神经元分化期间，SAMEN1的异紫色-1和异紫外线-2差异表达。通过阻断NGF信号传导的个体下游途径，我们发现PI3K / AKT途径在SALLIN1同种型-2的表达中起主要作用。蛋白质印迹和RT-PCR结果表明，SAMEN1的同种型-1组成型表达，而异紫色-2以严格依赖于NGF刺激的方式表达。异构体特异性RT-PCR结果表明，异紫外线-1和异紫外线-2的差异表达为SAOPIN1的结果是响应于NGF信号传导的SHOWIN1前mRNA的替代剪接。总的来说这些发现提供了调节Shootin1基因表达的分子机制的第一信息，并代表NGF诱导的替代剪接过程的第一个例子，其在神经发生中具有调节作用。

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