DNA Microarray Analysis of Neuronal Excitability

描述(由申请人提供)：这项建议的目标是建立神经兴奋性的遗传图谱，因为它与精神刺激性成瘾有关。为了实现这一目标，我们将使用最先进的寡核苷酸高密度微阵列技术和膜片钳电生理学相结合。Affymetrix基因芯片允许对整个小鼠基因组进行全基因组扫描，以寻找对长期服用精神刺激剂敏感的基因。为了理解大脑奖赏回路中与成瘾有关的神经信息处理，有必要了解该系统中的单个神经元是如何编码奖赏相关信息的。本项目利用DNA微阵列和膜片钳电生理学，对反复注射精神运动兴奋剂的大鼠和小鼠的伏隔核、前额叶皮质锥体神经元和腹侧下丘脑区锥体神经元的分子和功能特征进行了研究。具体目标1是寻找伏隔核、下丘脑和前额叶皮质神经元中受精神刺激剂调节的电压和配体门控离子通道基因。具体目标2是确定这些变化在每个结构中的功能表现。具体目标3涉及电生理分类神经元的单个神经元基因表达谱(例如，中间神经元和锥体爆发区和非爆发区)。总之，这些实验将帮助我们理解滥用药物如何在大脑奖励回路中诱导长期的神经适应。随着对成瘾相关可塑性的更多了解，有可能开发药理学或基因图谱策略来筛选成瘾易感性和可能的治疗干预措施。

DESCRIPTION (provided by applicant): The goal of this proposal is to establish a genetic profile of neuronal excitability as it relates to psychostimulant addiction. To accomplish this aim we will be using the most advanced oligonucleotide high density microarray technology combined with patch-clamp electrophysiology. Affymetrix Gene Chips allow for genome wide scans of the entire mouse genome for genes susceptible to chronic psychostimulant administration. In order to understand neural information procession in the brain reward circuitry as it relates to addiction, it is necessary to understand how individual neurons within this system code reward-related information. This project uses both DNA microarrays and patch-clamp electrophysiology to establish a molecular and functional characterization of nucleus accumbens, prefrontal cortical pyramidal neurons and ventral subicular hippocampal pyramidal neurons after repeated psychomotor stimulant injections in rats and mice. Specific aim 1 is to look for voltage and ligand-gated ion-channel genes that are subject to modulation by psychostimulants in the accumbens, subiculum and prefrontal cortical neurons. Specific aim 2 is to determine the functional manifestations of those changes within each structure. Specific aim 3 involves individual neuron gene expression profiling of electrophysiologically classified neurons (e.g. Interneurons and pyramidal bursting and nonbursting). Together these experiments will help us understand how drugs of abuse induce long-lasting neuroadaptations within the brain reward circuitry. With greater understanding of addiction-associated plasticity it may be possible to develop pharmacological or gene profiling strategies to screen for addiction vulnerability and possible therapeutic interventions.