RNA sequencing of poly(A+) transcriptome of quadriceps muscles from wildtype and CaMK2gVV/CaMK2gVV mice under sedentary and post-acute exercise conditions

Reactive oxygen species (ROS) contribute to health and disease. CaMKII is a widely expressed enzyme whose activation by oxidation of regulatory domain methionines (ox-CaMKII) contributes to cardiovascular disease, asthma, and cancer. Here we integrate comparative genomic and experimental data to show that CaMKII activation by ROS arose more than half-a-billion years ago on the vertebrate stem lineage where it constituted a bridge between ROS and increased intracellular Ca2+ release, exercise responsive gene transcription, and improved performance in skeletal muscle. These enhancements to fight-or-flight physiology were likely key in facilitating a well-evidenced shift in the behavioural ecology of our immediate chordate ancestors, and, in turn, the evolutionary success of vertebrates. Still, the ox-CaMKII innovation must be considered a critical evolutionary trade-off, as it rendered us more susceptible to common and often fatal diseases by providing ROS an expanded role in those processes. This poly(A+) transcriptome profiling study examined the acute transcriptional response of mouse skeletal muscles three hours after a single bout of submaximal exericse in wildtype and mutant mice whose CaMKIIg cannot be activated by reactive oxygen species. Wildtype and CaMK2gVV/CaMK2gVV mice were either kept sedentary or forced to performed a submaximal running exercise. Four mice per genotype were used for each condition. Three hours after the group of exercised mice finished running, all mice were sacrificed for mRNA sequencing by Illumina NovaSeq 6000.活性氧（Reactive oxygen species，ROS）参与机体健康与疾病进程。钙调蛋白依赖性蛋白激酶II（CaMKII）是一种广泛表达的酶，其通过调控结构域甲硫氨酸氧化而激活的氧化型钙调蛋白依赖性蛋白激酶II（ox-CaMKII）可引发心血管疾病、哮喘与癌症。本研究整合比较基因组学与实验数据，证实ROS介导的CaMKII激活起源于超过5亿年前的脊椎动物干演化支系，该机制充当了ROS与细胞内钙离子释放增加、运动应答基因转录以及骨骼肌功能改善之间的桥梁。这些对“战斗或逃跑”应激生理机能的增强，可能是推动我们近缘脊索动物祖先的行为生态学发生已有充分证据支持的转变的关键因素，进而助力脊椎动物的演化成功。然而，ox-CaMKII的演化革新需被视为一次关键的演化权衡：它通过扩大ROS在相关生理过程中的作用，使人类更易罹患常见且往往致命的疾病。本poly(A)+转录组谱分析（poly(A)+ transcriptome profiling）研究探究了单次亚最大强度运动3小时后，野生型与CaMKIIγ无法被活性氧激活的突变型小鼠的骨骼肌急性转录应答。将野生型及CaMK2gVV/CaMK2gVV小鼠分为静息组与强迫亚最大强度跑台运动组，每种基因型每组使用4只小鼠。运动组小鼠完成跑步训练3小时后，对所有小鼠实施安乐死，采集样本通过Illumina NovaSeq 6000平台进行mRNA测序。