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脑源性神经营养因子可提高培养的大鼠皮质神经元中 hnRNPs Q 和 R 的轴突水平。

BDNF elevates the axonal levels of hnRNPs Q and R in cultured rat cortical neurons.

机构信息

Institute of Molecular Medicine, National Tsing Hua University, Hsinchu, Taiwan.

Institute of Systems Neuroscience, National Tsing Hua University, Hsinchu, Taiwan.

出版信息

Mol Cell Neurosci. 2019 Jul;98:97-108. doi: 10.1016/j.mcn.2019.06.005. Epub 2019 Jun 14.

Abstract

Local translation plays important roles in the maintenance and various functions of axons, and dysfunctions of local translation in axons are implicated in various neurological diseases. Heterogeneous nuclear ribonucleoproteins (hnRNPs) are RNA binding proteins with multiple functions in RNA metabolism. Here, we identified 20 hnRNPs in the axons of cultured rat cortical neurons by interrogating published axon mass spectrometric databases with rat protein databases. Among those identified in axons are highly related hnRNPs Q and R. RT-PCR analysis indicated that axons also contained low levels of hnRNPs Q and R mRNAs. We further found that BDNF treatments raised the levels of hnRNPs Q and R proteins in whole neurons and axons. BDNF also increased the level of poly(A) RNA as well as the proportion of poly(A) RNA granules containing hnRNPs Q and R in the axon. However, following severing the connection between the cell bodies and axons, BDNF did not affect the levels of hnRNPs Q and R, the content of poly(A) RNA, or the colocalization of poly(A) RNA and hnRNPs Q and R in the axon any more, although BDNF still stimulated the local translation in severed axons as it did in intact axons. The results are consistent with that BDNF enhances the axonal transport of RNA granules. The results further suggest that hnRNPs Q and R play a role in the mechanism underlying the enhancement of axonal RNA transport by BDNF.

摘要

局部翻译在轴突的维持和各种功能中发挥重要作用,轴突中局部翻译的功能障碍与各种神经疾病有关。异质核核糖核蛋白 (hnRNPs) 是具有多种 RNA 代谢功能的 RNA 结合蛋白。在这里,我们通过用大鼠蛋白质数据库查询已发表的轴突质谱数据库,鉴定了培养的大鼠皮质神经元轴突中的 20 种 hnRNPs。在鉴定的轴突中,hnRNPs Q 和 R 高度相关。RT-PCR 分析表明,轴突中也含有低水平的 hnRNPs Q 和 R mRNA。我们进一步发现 BDNF 处理提高了整个神经元和轴突中 hnRNPs Q 和 R 蛋白的水平。BDNF 还增加了多聚(A)RNA 的水平以及含有 hnRNPs Q 和 R 的多聚(A)RNA 颗粒在轴突中的比例。然而,在切断细胞体和轴突之间的连接后,BDNF 不再影响 hnRNPs Q 和 R 的水平、多聚(A)RNA 的含量,或多聚(A)RNA 和 hnRNPs Q 和 R 在轴突中的共定位,尽管 BDNF 仍像在完整的轴突中一样刺激切断的轴突中的局部翻译。结果与 BDNF 增强 RNA 颗粒的轴突运输一致。结果进一步表明,hnRNPs Q 和 R 在 BDNF 增强轴突 RNA 运输的机制中起作用。

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