Centre for Brain Repair, University of Cambridge, Forvie Site, Cambridge CB2 0PY, United Kingdom.
RNA. 2011 Jan;17(1):85-98. doi: 10.1261/rna.2386111. Epub 2010 Nov 23.
mRNAs are transported, localized, and translated in axons of sensory neurons. However, little is known about the full repertoire of transcripts present in embryonic and adult sensory axons and how this pool of mRNAs dynamically changes during development. Here, we used a compartmentalized chamber to isolate mRNA from pure embryonic and adult sensory axons devoid of non-neuronal or cell body contamination. Genome-wide microarray analysis reveals that a previously unappreciated number of transcripts are localized in sensory axons and that this repertoire changes during development toward adulthood. Embryonic axons are enriched in transcripts encoding cytoskeletal-related proteins with a role in axonal outgrowth. Surprisingly, adult axons are enriched in mRNAs encoding immune molecules with a role in nociception. Additionally, we show Tubulin-beta3 (Tubb3) mRNA is present only in embryonic axons, with Tubb3 locally synthesized in axons of embryonic, but not adult neurons where it is transported, thus validating our experimental approach. In summary, we provide the first complete catalog of embryonic and adult sensory axonal mRNAs. In addition we show that this pool of axonal mRNAs dynamically changes during development. These data provide an important resource for studies on the role of local protein synthesis in axon regeneration and nociception during neuronal development.
mRNA 在感觉神经元的轴突中被运输、定位和翻译。然而,对于胚胎和成体感觉轴突中存在的完整转录本以及这些 mRNA 库在发育过程中如何动态变化,人们知之甚少。在这里,我们使用分隔室从纯净的胚胎和成体感觉轴突中分离 mRNA,这些轴突没有神经元或细胞体的污染。全基因组微阵列分析显示,大量以前未被识别的转录本定位于感觉轴突中,并且这个转录本谱在发育过程中向成年期发生变化。胚胎轴突富含与轴突生长有关的细胞骨架相关蛋白的编码转录本。令人惊讶的是,成年轴突富含编码免疫分子的 mRNA,这些分子在伤害感受中起作用。此外,我们还表明,Tubb3 mRNA 仅存在于胚胎轴突中,Tubb3 在胚胎轴突中局部合成,但在成年神经元中不合成,而是在成年神经元中运输,从而验证了我们的实验方法。总之,我们提供了第一个完整的胚胎和成体感觉轴突 mRNA 目录。此外,我们还表明,这个轴突 mRNA 库在发育过程中动态变化。这些数据为研究局部蛋白质合成在神经元发育过程中轴突再生和伤害感受中的作用提供了重要资源。