The Analysis of Central Neural Circuits by Subtractive Transgenics

描述（由申请人提供）：我们提出了一种方法来创建能够以解剖特异性方式稳健表达任何转基因的小鼠系。我们称这种方法为“减法转基因”，因为它涉及（通过两种已证实的转基因技术的结合）将一种表达模式的解剖特异性从另一种表达模式中减去。我们计划用这些小鼠解剖中枢神经系统（CNS）的功能电路，通过表达“沉默者”（关闭神经元的结构），具有前所未有的解剖特异性。这将允许我们和其他人以类似于工程师分析电子电路的方式来分析中枢神经系统的神经回路：短路一个元件，然后记录下游元件的情况。我们提出的前几个特定构建应该会导致前脑不同部位的转基因表达，这些部位与阿尔茨海默病和其他痴呆症（海马体形成）、帕金森氏病和其他基底神经节（纹状体）疾病等各种神经病理有关。由此产生的小鼠对中枢神经系统功能回路的深入了解将使我们更好地了解其病理状态的病因，并允许产生更好的人类疾病小鼠模型。然而，应该强调的是，该方法可以提高转基因在任何组织类型中表达的解剖特异性，因此应该对一般的生物医学研究有用。公共卫生相关性。哺乳动物的大脑是自然界中解剖学上最复杂的结构（而我们拥有最复杂的哺乳动物大脑），由成千上万种不同类型的细胞之间无数的电相互作用组成。从本质上讲，这是一个极其复杂的生物回路，细胞类型是它的组成部分。许多神经和精神疾病可以被认为是这种中央电路不同部分的不平衡。我们提出了一种方法来创建转基因小鼠系，这些小鼠系可以在这些中央回路的不同特定区域表达转基因，从而使研究人员能够尝试了解不同部分的作用。可以表达的转基因范围很广，从我们建议的关闭细胞的东西，到特定的遗传疾病模型。我们提出的前几个特定构建应该会导致前脑不同部位的转基因表达，这些部位与各种神经病理有关，如阿尔茨海默病和其他痴呆症（海马体形成）到帕金森病和其他基底神经节（纹状体）疾病。由此产生的小鼠对哺乳动物大脑功能回路的高度理解将使我们更好地了解其病理状态，并允许产生更好的人类疾病小鼠模型。

DESCRIPTION (provided by applicant): We propose a methodology to create lines of mice that are capable of the robust expression of any transgene in an anatomically specific manner. We call this method "subtractive transgenics", because it involves (by a combination of two proven transgenic technologies) the subtraction of the anatomical specificity of one expression pattern from that of another. We plan to use these mice to dissect out the functional circuitry of the central nervous system (CNS) by expressing "silencers" (constructs which turn neurons off) with unprecedented anatomical specificity. This should allow us and others to analyze the neural circuits of the CNS in a way analagous to how an engineer analyzes an electronic circuit: short out one element, and then record what happens to downstream elements. The first few specific constructs we propose should result in transgene expression in various parts of the forebrain that have been implicated in neuropathologies as diverse as Alzheimer's disease and other dementiae (the hippocampal formation) to Parkinson's disease and other disorders of the basal ganglia (striatum). The heightened understanding of the functional circuitry of the CNS the resulting mice will enable will lead to a better understanding of the etiology of its pathological states, and allow for the generation of better mouse models of these human disorders. However, it should be stressed that the method can increase the anatomical specificity of transgene expression in any tissue type, and should therefore be useful to biomedical research in general. PUBLIC HEALTH RELEVANCE. The mammalian brain is the most anatomically-complex structure in nature (and we have the most complex mammalian brain), composed of innumerable electrical interactions between literally thousands of different cell types. It is an incredibly complex biological circuit, in essence, and the cell types are its component parts. Many neurological and psychiatric disorders can be thought of as imbalances in different parts of this central circuitry. We propose a method to create genetically-modified lines of mice that can express transgenes in different specific areas of these central circuits, to enable researchers to try to understand what the different parts do. The transgenes that can be expressed range from things that just turn the cells off, as we propose to do, to specific genetic disease models. The first few specific constructs we propose should result in transgene expression in various parts of the forebrain that have been implicated in neuropathologies as diverse as as Alzheimer's disease and other dementiae (the hippocampal formation) to Parkinson's disease and other disorders of the basal ganglia (striatum). The heightened understanding of the functional circuitry of the mammalian brain the resulting mice will enable will lead to a better understanding of its pathological states, and allow for the generation of better mouse models of human disorders.