GluN3A基因缺失影响小鼠抑郁样行为及内侧前额叶皮质与海马mRNA转录组的实验研究

Genetic deletion of GluN3A affects depression-like behaviors and mRNA transcriptome in medial prefrontal cortex and hippocampus in mice

目的 探究GluN3A基因缺失参与抑郁样行为的潜在分子机制。方法 纳入成年雄性GluN3A基因敲除(knockout,KO)小鼠14只和同窝出生野生型(wild type,WT)小鼠15只,糖水偏好实验和强迫游泳实验后收集小鼠(每组5只)内侧前额叶皮质(medial prefrontal cortex,mPFC)和海马组织进行mRNA转录组测序,以DESeq2软件筛选差异表达基因并选取部分基因以qPCR进行验证,应用clusterProfiler软件对差异基因进行基因本体(Gene Ontology,GO)富集分析,应用string在线数据库和Cytoscape软件进行蛋白互作网络分析。结果 与WT小鼠相比,GluN3A KO小鼠糖水偏好(P<0.05)和糖水消耗量(P<0.05)显著下降,强迫游泳不动时间显著增加(P<0.01);在GluN3A KO小鼠mPFC和海马组织中分别筛选出740个和666个差异基因;GO分析显示,mPFC中下调差异基因显著富集在血管生成、神经前体细胞的增殖等条目,海马中下调差异基因主要富集在突触组装、轴突发育、树突发育、轴突引导、突触膜等条目,mPFC和海马中上调差异基因均主要与胞质核糖体和核糖体亚基有关;蛋白互作网络分析显示mPFC和海马中核心基因大部分为核糖体蛋白基因。结论 GluN3A基因缺失可能通过调控血管生成、神经前体细胞增殖、突触形成及蛋白合成等途径参与抑郁样行为的发生。

Objective To explore the potential molecular mechanism of GluN3A gene deletion involved in depression-like behaviors.Methods Fourteen male adult GluN3A knockout(KO) mice and 15 male wild type(WT) mice were recruited in this study.Sucrose preference test(SPT) and forced swim test(FST) were conducted to measure depression-like behaviors.Then tissue samples of medial prefrontal cortex(mPFC) and hippocampus(HP) were collected from 5 mice in each group for mRNA transcriptome sequencing.Differentially expressed genes(DEGs) were screened using DESeq2 software,and some DEGs were selected and validated by qPCR.Gene Ontology(GO) enrichment analysis was performed using clusterProfiler software.Protein interaction network analysis was performed using string online database together with Cytoscape software.Results GluN3A KO mice exhibited decreased sucrose preference(P<0.05) and sucrose consumption(P<0.05) in FST and prolonged immobile time(P<0.01) in FST.There were 740 and 666 DEGs found in the mPFC and the hippocampus of GluN3A KO mice,respectively.GO analysis showed that down-regulated DEGs in the mPFC were enriched in angiogenesis and neural precursor cell proliferation,while down-regulated DEGs in the hippocampus were mainly enriched in synapse assembly,axon development,dendrite development,axon guidance and synaptic membrane.And up-regulated DEGs in both mPFC and hippocampus were mainly associated with cytosolic ribosome and ribosomal subunit.Protein interaction network analysis revealed that most of the core genes in the mPFC and hippocampus were ribosomal protein genes.Conclusion GluN3A deletion may be involved in the occurrence of depression-like behaviors by regulating angiogenesis,neural precursor cell proliferation,synapse formation and protein synthesis.