CA1中央区NMDA受体G1uN3A亚基参与应激致抑郁和D-serine杭抑郁的细胞和环路机制
文献类型：学位论文
| 作者 | 张伟
|
| 答辩日期 | 2023-12 |
| 文献子类 | 博士 |
| 授予单位 | 中国科学院大学 |
| 授予地点 | 中国科学院心理研究所 |
| 其他责任者 | 王玮文 |
| 关键词 | 抑郁相关行为 NMDA受体G1uN3A亚基 海马CA1 锥体神经元 D一丝氨酸 |
| 学位名称 | 理学博士 |
| 学位专业 | 健康心理学 |
| 其他题名 | Cellular and circuit mechanisms involving CA1 intermediate area NMDA receptor G1uN3A subunit instress-induced denression and D-serine antidepressant action |
| 中文摘要 | Objective: Depression is a globally prevalent severe mental disorder, imposing substantial psychological, physiological, and economic burdens on individuals and society. Altered structure and function of N-methyl-D-aspartic acid (NMDA) receptors are known to be a critical molecular basis for the occurrence and treatment of depressive behaviors. G1uN3A, a subunit of the NMDA receptor, plays a crucial role in brain development and function related to NMDA receptor activity and has been implicated in various psychiatric disorders. However, the understanding of the relationship between G1uN3A subunits and depression remains limited. This study aims to explore the key role of G1uN3A subunits in the occurrence and treatment of depression, along with the underlying neural basis, providing new insights into the pathophysiology of depression and rapid antidepressant therapy. Methods: A mouse model of depression induced by chronic social defeat stress and transgenic mice with G1uN3A knockout (Grin3A-/一)were employed to investigate the relationship and mechanisms of G1uN3A subunits in depression-related behaviors. Firstly, the relationship between depression-related behaviors and changes in G1uN3A subunit expression in emotion-related brain areas was assessed through systematic behavioral and molecular biology tests. Secondly, in the hippocampal subregion with significant gene expression changes, the causal relationship between G1uN3A subunit expression and depression-related behaviors was established by locally injecting adeno-associated virus (AAV) vectors to selectively regulate G1uN3A subunit expression. Thirdly, the neurocellular and neural circuit basis of G1uN3A subunit regulation of depression-related behaviors in the hippocampal subregion was explored using a combination of immunohistochemistry, chemogenetic regulation, and tract tracing. Finally, the pharmacological role of G1uN3A subunits in the mediation of D-serine antidepressant effects was investigated through brain ventricular administration, chemogenetic regulation, and immunohistochemistry. Results: 1) Chronic social defeat stress induced depression-related behaviors in mice, including social interest loss, increased helpless behavior, and elevated anxiety behavior. In the emotion-related brain area, particularly the CA1 intermediate area (CAli) of the hippocampus, the mRNA and protein expression levels of G1uN3A subunits were significantly downregulated. Similarly, G1uN3A knockout mice exhibited a phenotype of depression-related behaviors, specifically increased helpless behavior, while social interest and anxiety-like behavior remained unchanged. These results suggest an association between insufficient G1uN3A subunit expression in the brain, particularly in the CAli, and depression-related behaviors, especially helpless behavior. 2) Further, the reduction of reduced G1uN3A subunit expression in the CAli of the hippocampus, achieved by locally injecting AAV virus to restore G1uN3A subunit expression, selectively reversed stress-induced depression-related helpless behavior. However, it lacked an effect on reduced social interest and anxiety behavior. Similarly, restoring G1uN3A subunit expression in the CAli of G1uN3A knockout mice using AAV virus also reversed their helpless behavior, indicating that the CAli of the hippocampus is a crucial site for G1uN3A subunit regulation of depression-related helpless behavior. 3) Further investigation of changes in the activity of CAli region neurons in response to stress, after the helpless behavior test, revealed that stress primarily led to a significant decrease in c-Fos-positive cells in pyramidal neurons of this region. This decrease could be restored by upregulating G1uN3A subunit expression in the CAli, indicating that G1uN3A subunits regulate the activity of pyramidal neurons in this region. In addition, during the helpless behavior test, chemogenetic tools (AAV-hM3Dq) were used to enhance the activation of CAli neurons, which reversed stress-induced depression-related helpless behavior. Conversely, inhibiting the activation of CAli neurons in normal animals through chemogenetic tools (AAV-hM4Di) directly led to an increase in helpless behavior. These results suggest that the functional activity of pyramidal neurons in the CAli region is the neural cellular basis for G1uN3A subunit regulation of depression-related helpless behavior. 4) Through tract tracing of CAli neurons' projection brain areas, it was found that they projected to emotion-related brain areas, including CA3, the Nucleus Accumbens (NAC), Basal Lateral Amygdala (BLA), Infralimbic Cortex (IL), and Lateral Entorhinal Cortex (LEC). Subsequent analysis of the number of c-Fos-positive cells in these projection brain areas after the helpless behavior test revealed a significant decrease in CA3 and IL brain areas due to stress, and this decrease was restored after upregulating G1uN3A subunit expression in the CAli. This suggests that the neural projections of CAli to CA3 and IL may be the circuit basis for G1uN3A subunit regulation of depression-related helpless behavior. 5) Local microinjection of the endogenous coagonist of NMDA receptors, D-serine, into the CAli region significantly improved stress-induced depression-related helpless behavior and increased the number of c-Fos-positive cells in CAli pyramidal neurons. Conversely, blocking the activation of CAli pyramidal neurons using chemogenetic tools (AAV-hM4Di) prevented the antidepressant effect of D-serine on the above depression-related behaviors and molecular changes. Additionally, D-serine intervention failed to reverse helpless behavior in G1uN3A knockout model animals and did not significantly increase the number of c-Fos-positive cells in CAli pyramidal neurons. These results indicate that the functional activity of CAli pyramidal neurons mediates the antidepressant effect of D-serine, and this effect is mediated by G1uN3A subunits. Conclusion: The results of this study reveal that the NMDA receptor G1uN3A subunit plays a crucial role through CAli pyramidal neurons and circuits in depression-related helpless behavior and the rapid antidepressant effect of D-serine. This provides important insights into the complex pathophysiology of depression and offers new perspectives and potentially transformative targets for rapid antidepressant therapy. |
| 英文摘要 | 目的:抑郁症是一种在全球范围内高发的严重精神障碍，对患者及社会造成了巨大的心理、生理和经济负担。已知N-甲基-D-天冬氨酸(N-methyl-D-asparticacid, NMDA)受体结构和功能的改变在抑郁行为的发生和治疗过程中是一种关键的分子基础。G1uN3A是组成NMDA受体的亚基团的一种，其在NMDA受体功能相关的脑发育和功能中扮演有重要角色，并被发现可能与多种精神疾病有关。然而G1uN3A亚基与抑郁症之间关系的了解仍十分匮乏。本研究旨在探讨G1uN3A亚基在抑郁发生和治疗中的关键作用及其背后所参与的神经基础，为深化抑郁症病理生理的认识和快速抗抑郁治疗提供新的见解。 方法:采用慢性社会挫败应激诱导的小鼠抑郁模型和G1uN3A敲除C Grin3A一的转基因模式小鼠来探讨G1uN3A亚基与抑郁相关行为的关系和作用机制。首先，通过系统的行为学和分子生物学检测评估抑郁相关行为与情绪相关脑区G1uN3A亚基表达改变的关系。其次，在存在显著基因表达改变的海马亚脑区，通过脑立体手术局部微注射腺相关(Adeno-associated virus, AAV)病毒特异性调控G1uN3A亚基表达以确立其与抑郁相关行为的因果关系。第三，采用免疫组织化学、化学遗传调控和束路追踪相结合的方法探究海马亚区G1uN3A亚基对抑郁相关行为调控作用的神经细胞和神经环路基础。最后，通过脑部套管给药、化学遗传调控和免疫组织化学相结合的手段对海马亚区G1uN3A亚基在D-serine抗抑郁中的介导作用进行药理学探究。 结果:1)慢性社会挫败应激诱导了小鼠的抑郁相关行为，包括社交兴趣缺失、无助行为增加以及焦虑行为上升，且情绪相关脑区中，主要是海马的CA1中央区(CA1 intermediate area, CAli) G1uN3A亚基的mRNA和蛋白表达水平显著下调;采用G1uN3A亚基敲除(knockout, KO)小鼠同样发现该亚基的缺失会出现抑郁相关行为表型，具体表现为无助行为增加，而社交兴趣和焦虑样行为不变。这些结果表明脑中G1uN3A亚基表达的不足与抑郁行为，尤其是无助行为相关。 2)进一步通过微注射AAV病毒恢复海马CAli G1uN3A亚基表达的降低可以选择性逆转应激诱导的抑郁相关无助行为，但对社交兴趣降低和焦虑行为缺乏影响。同时通过AAV病毒特异性恢复G1uN3A KO小鼠海马CAli G1uN3A亚基的表达，也逆转了其无助行为，提示海马CAli是G1uN3 A亚基调控抑郁相关无助行为的关键部位。 3)进一步在无助行为检测后对应激致抑郁模型动物CAli区域神经元的活动变化进行考察，发现应激主要导致该区域锥体神经元中神经元活动标记物c-Fos阳性细胞数显著降低，且该降低可以通过上调CAli中的G1uN3A亚基表达恢复，提示G1uN3A亚基调控了该区域锥体神经元的活动。而在进行无助行为测试时，通过化学遗传工具AAV-hM3Dq增加CAli神经元的激活，可以逆转应激致抑郁模型动物的无助行为;相反地，通过AAV-hM4Di抑制正常动物的CAli神经元的激活，则可以直接导致无助行为的增加。这些结果表明CAli区域锥体神经元的功能活性是 G1uN3A亚基调控抑郁相关无助行为的神经细胞基础。 4)通过束路追踪CAli神经元的投射脑区，发现其投射至CA3、伏隔核(Nucleus accumbens, NAC、基底外侧杏仁核(Basal lateral amygdala, BLA)、边缘下区(Infralimbic cortex, IL)和外侧内嗅皮层(Lateral entorhinal cortex, LEC等情绪相关脑区。随后对这些投射脑区在无助行为检测后的c-Fos阳性细胞数量进行分析发现，应激导致CA3和IL脑区c-Fos阳性细胞数显著下降，并在CAliG1uN3A亚基上调后恢复，表明CA 1 i-CA3和CA 1 i-IL的神经投射可能是G1uN3A亚基调控抑郁相关无助行为的环路基础。 5 ) CAli区域局部微注射NMDA受体的内源性共激动剂D一丝氨酸 C D-serine)可以快速改善应激致抑郁模型动物的无助行为，同时提高了CAli锥体神经元中的c-Fos阳性细胞数量;而通过AAV-hM4Di同步抑制CAli锥体神经元激活则阻断了D-serine的抗抑郁效果。另外，D-serine干预无法逆转G1uN3A KO模式动物的无助行为，也无法显著提高CAli锥体神经元中c-Fos阳性细胞的数量。这些结果表明CAli锥体神经元功能活性介导了D-serine的抗抑郁作用，且该作用是G1uN3A亚基介导的。 结论:本研究结果揭示了NMDA受体G1uN3A亚基通过CAli锥体神经元及环路在抑郁相关无助行为和D-serine的快速抗抑郁中发挥重要的作用，为抑郁症的复杂病理机制提供了新观点，并为快速抗抑郁治疗提供了新思路和具有转化潜力的新靶点。 |
| 语种 | 中文 |
| 源URL | [http://ir.psych.ac.cn/handle/311026/46617]  |
| 专题 | 心理研究所_健康与遗传心理学研究室 |
| 推荐引用方式 GB/T 7714 | 张伟. CA1中央区NMDA受体G1uN3A亚基参与应激致抑郁和D-serine杭抑郁的细胞和环路机制[D]. 中国科学院心理研究所. 中国科学院大学. 2023. |
入库方式： OAI收割
来源：心理研究所
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