The neuromuscular junction (NMJ) is a prototypic chemical synapse between the spinal motor neuron and the motor endplate. Gene expression profiles of the motor endplate are not fully elucidated. Collagen Q (ColQ) is a collagenic tail subunit of asymmetric forms of acetylcholinesterase and is driven by two distinct promoters. pColQ1 is active throughout the slow-twitch muscle, whereas pColQ1a is active at the motor endplate of fast-twitch muscle. We made a transgenic mouse line that expresses nuclear localization signal (NLS)-attached Cre recombinase under the control of pColQ1a (pColQ1a-Cre mouse). RiboTag mouse expresses an HA-tagged ribosomal subunit, RPL22, in cells expressing Cre recombinase. We generated pColQ1a-Cre:RiboTag mouse, and confirmed that HA-tagged RPL22 was enriched at the NMJ of tibialis anterior (TA) muscle. Next, we confirmed that Chrne and Musk that are specifically expressed at the NMJ were indeed enriched in HA-immunoprecipitated (IP) RNA, whereas Sox10 and S100b, markers for Schwann cells, and Icam1, a marker for vascular endothelial cells, and Pax3, a marker for muscle satellite cells, were scarcely detected. Gene set enrichment analysis (GSEA) of RNA-seq data showed that “phosphatidylinositol signaling system” and “extracellular matrix receptor interaction” were enriched at the motor endplate. Subsequent analysis revealed that genes encoding diacylglycerol kinases, phosphatidylinositol kinases, phospholipases, integrins, and laminins were enriched at the motor endplate. We first characterized the gene expression profile under translation at the motor endplate of TA muscle using the RiboTag technique. We expect that our gene expression profiling will help elucidate molecular mechanisms of the development, maintenance, and pathology of the NMJ.

神经肌肉接头（NMJ）是脊髓运动神经元和运动终板之间的原型化学突触。尚未完全阐明电机终板的基因表达谱。胶原蛋白Q（ColQ）是不对称形式的乙酰胆碱酯酶的胶原蛋白尾部亚基，由两个不同的启动子驱动。p考1 在整个慢肌中活跃，而p甲型在快速抽搐肌肉的运动终板上活跃。我们制作了一个在p的控制下表达核定位信号（NLS）附着的Cre重组酶的转基因小鼠品系甲型 （p甲型-Cre鼠标）。RiboTag小鼠在表达Cre重组酶的细胞中表达HA标记的核糖体亚基RPL22。我们产生了p甲型-Cre：RiboTag小鼠，并确认HA标签的RPL22在胫前肌（TA）肌肉的NMJ处富集。接下来，我们确认r 和 麝香 在NMJ上特异性表达的那些确实富含HA免疫沉淀（IP）RNA，而 袜10 和 S100b，雪旺细胞标记和 Icam1，是血管内皮细胞的标记，以及 Pax3，几乎没有检测到肌卫星细胞的标志物。RNA-seq数据的基因集富集分析（GSEA）表明，“磷脂酰肌醇信号系统”和“细胞外基质受体相互作用”在运动终板上富集。随后的分析表明，编码二酰基甘油激酶，磷脂酰肌醇激酶，磷脂酶，整联蛋白和层粘连蛋白的基因在运动终板上富集。我们首先使用RiboTag技术表征了TA肌肉运动终板在翻译下的基因表达谱。我们希望我们的基因表达谱分析将有助于阐明NMJ发生，维持和病理的分子机制。