DNA Microarray Analysis of Neuronal Excitability

描述（由申请人提供）：本提案的目标是建立神经元兴奋性的遗传图谱，因为它与精神兴奋剂成瘾有关。为了实现这一目标，我们将使用最先进的寡核苷酸高密度芯片技术结合膜片钳电生理学。Affyphin基因芯片允许对整个小鼠基因组进行基因组范围的扫描，以寻找对慢性精神兴奋剂给药敏感的基因。为了理解大脑奖励回路中与成瘾相关的神经信息处理，有必要了解该系统中的单个神经元如何编码与奖励相关的信息。本研究利用DNA微阵列技术和膜片钳电生理技术，对大鼠和小鼠反复注射精神兴奋剂后的丘脑核、前额叶皮质锥体神经元和腹侧海马下托锥体神经元进行了分子和功能表征。具体目标1是寻找电压和配体门控离子通道基因，受到调制的精神兴奋剂，下托和前额叶皮层神经元。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涉及电生理学分类的神经元（例如中间神经元和锥体爆发和非爆发）的个体神经元基因表达谱。这些实验将帮助我们了解滥用药物如何在大脑奖励回路中诱导持久的神经适应。随着对成瘾相关可塑性的进一步了解，可能会开发药理学或基因分析策略来筛选成瘾脆弱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.