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学特异性方式稳健表达任何转基因的小鼠品系。 我们称这种方法为“减法转基因”，因为它涉及（通过两种成熟的转基因技术的组合）从另一种表达模式中减去一种表达模式的解剖特异性。 我们计划使用这些小鼠通过表达具有前所未有的解剖特异性的“沉默器”（关闭神经元的结构）来解剖中枢神经系统（CNS）的功能电路。 这将使我们和其他人能够分析中枢神经系统的神经回路，就像工程师分析电子回路一样：使一个元件短路，然后记录下游元件发生的情况。 我们提出的前几个特定的构建体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（海马结构）、帕金森病和基底神经节（纹状体）其他疾病等不同的神经病理学有关。 由此产生的小鼠对CNS的功能电路的进一步理解将使其能够更好地理解其病理状态的病因学，并允许产生这些人类疾病的更好的小鼠模型。 然而，应该强调的是，该方法可以增加在任何组织类型中转基因表达的解剖学特异性，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（我们有最复杂的哺乳动物大脑），由成千上万种不同细胞类型之间的无数电相互作用组成。 从本质上讲，它是一个非常复杂的生物回路，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个中央回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品系，这些小鼠可以在这些中央回路的不同特定区域表达转基因，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从只是关闭细胞的东西，正如我们所建议的那样，到特定的遗传疾病模型。 我们提出的前几个特定的构建体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（海马结构）到帕金森病和基底神经节（纹状体）的其他疾病等不同的神经病理学有关。 对哺乳动物大脑功能电路的深入了解将使小鼠能够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.