Glutamate receptors in pain and addictive behaviors

描述(由申请人提供)： 这是一项延长高级科学家奖的请求，这将使我能够继续获得分子遗传学和神经科学技术，并利用它们来实现NIDA资助的两个研究项目的目标。这两个项目都是为了检验一种假设，即由NR1或GluR2亚基组成的谷氨酸能受体对于神经元的可塑性是必需的，这种可塑性表现为损伤诱导的疼痛(伤害性感受)和反复使用可卡因或阿片类药物时发生的适应，并表现为药物奖励和复发以及阿片类药物的耐受和依赖。第一个项目将使用我们开发的时空基因敲除(KO)来确定NMDAR1(NMDA受体)和/或GluR2(AMPA受体)亚单位在炎症和脊髓神经损伤引起的持续性疼痛中的作用。第二个项目将使用同样的Cre-loxP技术来产生NMDA和AMPA受体的KO，该KO仅限于成年小鼠的特定脊髓上中枢神经系统区域，包括伏隔核、腹侧被盖区和腹侧下丘(空间KO)(TIMAL KO)。我们将通过免疫细胞化学、原位杂交和电子显微镜技术，展示与靶受体亚单位缺失相关的基因表达变化的空间定位和时间特征。我们还将确定，在上述选定的中枢神经系统区域定向删除这些受体亚基是否会减弱谷氨酸能电流，减少或阻止对可卡因和阿片类药物使用的行为适应，包括药物奖励和复发以及阿片耐受和依赖。 这些技术的获取和应用将通过协作安排和利用补充我实验室现有专门知识的机构核心设施来实现。 这项SSA奖还将使我继续减少教学和行政责任，这大大增加了我能够致力于提高我的科学技能的时间。

DESCRIPTION (provided by applicant): This is a request for a renewal of a Senior Scientist Award that will allow me to continue to acquire molecular genetic and neuroscience techniques and to use them to achieve the goals of two NIDA funded research projects. Both projects are designed to test the hypothesis that glutamatergic receptors composed of NR1 or GluR2 subunits are necessary for the neuronal plasticity that is manifest as injury-induced pain (nociception) and the adaptations that occur during repeated cocaine or opioid use and are manifest as drug reward and relapse as well as opioid tolerance and dependence. The first project will use the spatial-temporal knockout (KO) that we have developed to determine the role of spinal cord dorsal horn of NMDAR1 (NMDA receptor) and/or GluR2 (AMPA receptor) subunits in persistent pain resulting from inflammation and spinal nerve injury. The second project will use this same Cre-loxP technology to produce a KO of the NMDA and AMPA receptors that is confined to selected supraspinal CNS areas including the nucleus accumbens, ventral tegmental area and ventral subiculum (a spatial KO) in adult mice (a temporal KO). We will demonstrate by immunocytochemical, in situ hybridization and electron microscopy techniques, the spatial localization and temporal characteristics of the changes in gene expression that are associated with the deletion of the target receptor subunits. We will also determine whether the targeted deletion of these receptor subunits in the selected CNS areas given above will attenuate glutamatergic currents and decrease or prevent the behavioral adaptations to cocaine and opioid use including drug reward and relapse and opioid tolerance and dependence. The acquisition and application of these techniques will be accomplished through collaborative arrangements and through the utilization of institutional core facilities that complement the expertise available in my laboratory. This SSA award will also allow me to continue the reduction in teaching and administrative responsibilities that has significantly increased the time I was able to devote to enhancing my scientific skills.