工具病毒系统的建立及其在神经环路解析中的应用

发布时间：2018-03-22 17:15

本文选题：嗜神经病毒　切入点：情绪和记忆　出处：《中国科学院大学(中国科学院武汉物理与数学研究所)》2016年博士论文　论文类型：学位论文

【摘要】：绘制大脑神经网络的连接图谱有助于我们更好地解析大脑的功能,发展用于绘制大脑神经网络结构的技术,已成为当前神经科学研究的迫切需求。传统的神经环路研究的方法主要包括高尔基染色以及蛋白质或染料标记的方法。基于病毒发展而来的神经环路示踪技术,是目前绘制大脑连接图谱的常用手段之一,而且也是近年来神经科学研究中发展十分迅速的领域。目前常用的用于标记神经环路的嗜神经病毒主要有伪狂犬病毒(pseudorabies virus,PRV)、重组狂犬病毒(rabiesvirus,RV)、水疱性口炎病毒(vesicularstomatitisvirus,VSV)和单纯疱疹病毒(herpes simplex virus,HSV),这些嗜神经病毒可以用于解析大脑中枢特定核团或者外周器官的单级或多级的输入输出网络。另外一些常用的解析神经环路的病毒包括腺相关病毒(adeno-associatedvirus,AAV)等病毒,可以用于神经环路的操控以及神经元投射等研究。利用上述这些病毒可以解析目前神经科学研究中的部分问题,本研究利用上述病毒标记了海马和杏仁核的环路以及神经元的精细形态的描绘。主要内容是:1、协助实验室建立并完善了用于解析神经环路的多种病毒系统。2、合作研究完成习得性绝望和习得性充满希望两种情感对小鼠空间记忆的不同的影响的神经环路机制。通过对比不同情绪状态下,小鼠空间学习记忆能力的变化,从而鉴定出基底外侧杏仁核靠后部(BLP)和腹侧海马CA1的单突触连接调控了情绪对空间学习记忆能力的影响,该部分工作主要与华中科技大学同济医学院王建枝教授课题组合作完成。3、合作研究完成了稀疏标记小鼠大脑神经元并追踪重构单个神经元的方法,用AAV病毒稀疏标记小鼠大脑多个脑区的神经元的形态并使用fMOST(华中科技大学光电国家实验室技术)对外侧前额叶进行精细结构成像,获得了小鼠外侧前额叶的稀疏标记的神经元的全脑投射数据。该部分工作主要是和华中科技大学光电国家实验室张玉慧教授课题组和曾绍群教授课题组合作完成。4、合作研究了小鼠海马区苔藓细胞(Mossy cell)的精细形态,主要方法是利用上述的可以用于稀疏标记的AAV病毒注射海马DG区,该病毒可以选择性地稀疏标记海马DG区的苔藓细胞,通过该方法可以获得苔藓细胞的精细形态结构,该部分工作主要是和华中科技大学光电国家实验室张玉慧教授课题组和曾绍群教授课题组合作完成。
[Abstract]:Mapping the connections of neural networks in the brain will help us better analyze the functions of the brain and develop techniques for mapping the network structure of the neural network of the brain. It has become an urgent need to study neuroscience. The traditional methods of neural loop research mainly include Golgi staining and protein or dye labeling. Neural loop tracing technology based on virus development. Is one of the most common methods of mapping brain connections. It is also a rapidly developing field of neuroscience research in recent years. Currently, the commonly used neuroviruses used to label neural loops are pseudorabies virus (PRV), Recombinant rabies virus (RV), vesicularis stomatitis virus (VSVV) and single rabies virus (RV). Herpes simplex virus.These neuroviruses can be used to parse single-stage or multi-stage input-output networks in specific nuclei or peripheral organs of the brain. Other commonly used viruses that parse nerve loops include viruses such as adeno-associated virus, adeno-associated virus, or adeno-associated virus. These viruses can be used to study neural loop manipulation and neuronal projection. These viruses can be used to analyze some of the problems in current neuroscience research. This study used the virus to label the loop of hippocampus and amygdala and the description of fine morphology of neurons. The main content of this study is: 1, to assist the laboratory to establish and improve a variety of virus systems used to analyze the neural loop. To study the neural loop mechanism of the different effects of the two emotions on the spatial memory of mice, namely, learning despair and learning hopefulness. The changes of spatial learning and memory ability in mice showed that the effects of emotion on spatial learning and memory were regulated by the single synaptic connections between the basolateral amygdaloid nucleus and the ventral hippocampal CA1. This part mainly works with Wang Jianchi Group of Tongji Medical College of Huazhong University of Science and Technology to study the method of sparse labeling mouse brain neurons and tracking the reconstruction of single neurons. The morphology of neurons in multiple brain regions of mice was labeled sparsely with AAV virus and fine structure imaging of the lateral prefrontal lobe was performed using fMOST (Optoelectronic National Laboratory of Huazhong University of Science and Technology). The global brain projection data of sparsely labeled neurons in the lateral prefrontal lobe of mice were obtained. This part of the work is mainly done in collaboration with Professor Zhang Yuhui, National Laboratory of Optoelectronics, Huazhong University of Science and Technology, and Professor Zeng Shaoqun. 4. The fine morphology of mossy cells in the hippocampus of mice was studied. The main method is to inject the DG region of hippocampus with the above-mentioned AAV virus which can be used for sparse labeling. The virus can selectively and sparsely label the mossy cells in the DG region of the hippocampus. By this method, the fine morphology of the mossy cells can be obtained. The main work of this part is to cooperate with Professor Zhang Yuhui and Professor Zeng Shaoqun of National Laboratory of Optoelectronics of Huazhong University of Science and Technology (Huazhong University of Science and Technology).
【学位授予单位】：中国科学院大学(中国科学院武汉物理与数学研究所)
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R338

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