RNA Sequencing of Trigeminal Ganglia in <i>Rattus Norvegicus</i> after Glyceryl Trinitrate Infusion with Relevance to Migraine

Introduction Infusion of glyceryl trinitrate (GTN), a donor of nitric oxide, induces immediate headache in humans that in migraineurs is followed by a delayed migraine attack. In order to achieve increased knowledge of mechanisms activated during GTN-infusion this present study aims to investigate transcriptional responses to GTN-infusion in the rat trigeminal ganglia. Methods Rats were infused with GTN or vehicle and trigeminal ganglia were isolated either 30 or 90 minutes post infusion. RNA sequencing was used to investigate transcriptomic changes in response to the treatment. Furthermore, we developed a novel method for Gene Set Analysis Of Variance (GSANOVA) to identify gene sets associated with transcriptional changes across time. Results 15 genes displayed significant changes in transcription levels in response to GTN-infusion. Ten of these genes showed either sustained up- or down-regulation in the 90-minute period after infusion. The GSANOVA analysis demonstrate enrichment of pathways pointing towards an increase in immune response, signal transduction, and neuroplasticity in response to GTN-infusion. Future functional in-depth studies of these mechanisms are expected to increase our understanding of migraine pathogenesis.引言 硝酸甘油（glyceryl trinitrate, GTN）作为一氧化氮供体，其输注可使人体产生即刻头痛，偏头痛患者在头痛后还会继发延迟性偏头痛发作。为深入解析GTN输注过程中激活的分子机制，本研究旨在探究大鼠三叉神经节对GTN输注的转录应答反应。 方法：将大鼠分为GTN输注组与溶剂对照组，分别于输注后30分钟、90分钟采集三叉神经节组织。采用RNA测序（RNA sequencing）技术分析处理后样本的转录组变化。此外，本研究开发了一种全新的方差基因集分析（Gene Set Analysis Of Variance, GSANOVA）方法，用于识别不同时间点下与转录变化相关的基因集。 结果：共有15个基因的转录水平在GTN输注后出现显著变化，其中10个基因在输注后90分钟内呈现持续上调或下调趋势。GSANOVA分析结果显示，与GTN输注应答相关的通路显著富集于免疫应答、信号转导及神经可塑性相关通路。未来针对这些机制的深入功能研究，将有助于进一步阐明偏头痛的发病机制。