Zhang Ya-Qi, Zhang Qing, Yang Yi, Yu Li-Li, Fan Ning-Lin, Wu Yong, Wang Jun-Yang, Dang Xing-Lun, Guo Ying-Qi, Li Cong, Ma Guo-Lan, Wang Lu, Guo Yong-Bo, Li Shi-Wu
State Key Laboratory of Genetic Evolution & Animal Models, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, China.
Yunnan Key Laboratory of Animal Models and Human Disease Mechanisms, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, China.
Mol Psychiatry. 2025 May 17. doi: 10.1038/s41380-025-03052-7.
Single nucleotide polymorphisms (SNPs) within 1p31.1 region have shown significant associations with depression, and our prior functional genomics pinpointed a regulatory variant rs3101339 among them. However, its precise role in depression pathogenesis remains elusive. In this study, we employed a series of analytical and functional approaches, including regulatory element annotation, brain expression quantitative trait loci (eQTL), reporter gene assay, electrophoretic mobility shift assay (EMSA), and precise genome editing. Our results confirmed that rs3101339 is a causal variant within 1p31.1 with its risk allele C upregulating NEGR1 expression. To further investigate the consequences of NEGR1 upregulation, we overexpressed NEGR1 in specific region of the mouse brain (including medial prefrontal cortex (mPFC) and ventral hippocampus (vHIP)) using stereotaxic injection. Behavioral assessments revealed that elevated NEGR1 levels in the brain, particularly in the vHIP, resulted in working memory impairment as well as anxiety- and depression-like behaviors in mice. Neuronal sparse labeling assay and transmission electron microscopy revealed that NEGR1 overexpressing in the vHIP leads to dendritic spine loss and synaptic ultrastructure abnormality. Immunoprecipitation-mass spectrometry (IP-MS) further identified 67 high-confidence proteins that interacted with NEGR1, many of which are involved in neurotransmitter exocytosis and synaptic vesicle endocytosis. Transcriptomic profiling revealed 94 differentially expressed genes in NEGR1-OE (vHIP) mice compared to control mice (P adj < 0.05), which were enriched in myelination-related signaling pathways (such as myelination, ensheathment of neurons, axon ensheathment in central nervous system, etc.). Together, our findings implicated that the overexpression of the NEGR1 gene in the mouse brain as a potential driver of anxiety- or depression-like phenotypes potentially through impairing synaptic function and myelination.
1p31.1区域内的单核苷酸多态性（SNP）已显示出与抑郁症有显著关联，我们之前的功能基因组学研究在其中确定了一个调控变体rs3101339。然而，其在抑郁症发病机制中的精确作用仍不清楚。在本研究中，我们采用了一系列分析和功能方法，包括调控元件注释、脑表达定量性状基因座（eQTL）、报告基因检测、电泳迁移率变动分析（EMSA）和精确的基因组编辑。我们的结果证实，rs3101339是1p31.1区域内的一个因果变体，其风险等位基因C上调NEGR1的表达。为了进一步研究NEGR1上调的后果，我们使用立体定位注射在小鼠脑的特定区域（包括内侧前额叶皮质（mPFC）和腹侧海马体（vHIP））过表达NEGR1。行为评估显示，脑中NEGR1水平升高，特别是在vHIP中，导致小鼠工作记忆受损以及出现焦虑样和抑郁样行为。神经元稀疏标记分析和透射电子显微镜显示，在vHIP中过表达NEGR1会导致树突棘丢失和突触超微结构异常。免疫沉淀-质谱（IP-MS）进一步鉴定出67种与NEGR1相互作用的高可信度蛋白质，其中许多参与神经递质胞吐和突触小泡内吞作用。转录组分析显示，与对照小鼠相比，NEGR1过表达（vHIP）小鼠中有94个差异表达基因（P校正<0.05），这些基因富集于髓鞘形成相关信号通路（如髓鞘形成、神经元包被、中枢神经系统轴突包被等）。总之，我们的研究结果表明，小鼠脑中NEGR1基因的过表达可能通过损害突触功能和髓鞘形成，成为焦虑样或抑郁样表型的潜在驱动因素。
