Control of Cocaine Seeking by D1-dependent Modulation of NMDA Signaling

描述(由申请人提供)：Pavel Ortinski博士目前提交的为期五年的指导职业发展奖(K01)申请建议获得药物滥用行为药理学方面的培训。具体地说，所要求的培训将检查D1样多巴胺和NMDA受体相互作用在可卡因自我给药和恢复可卡因寻找的动物模型中的作用。这项培训将有助于 制定了一项独立的研究计划，重点是分析可卡因寻找和复发的突触机制。这项研究将在克里斯·皮尔斯博士的指导下在宾夕法尼亚大学进行。候选人：奥尔廷斯基博士在几个实验模型中展示了实施突触功能和生理学分析的研究项目的记录。他的博士论文在斯特凡诺·维奇尼博士的指导下在乔治城大学完成，利用细胞培养和脑片中的电生理膜片钳记录，研究了GABA-A受体亚型对抑制性突触强度的调节。Ortinski博士随后在费城儿童医院/宾夕法尼亚大学完成了与Douglas Coulter博士为期两年的博士后研究，在那里他评估了星形细胞谷氨酸/谷氨酰胺代谢途径在癫痫患者抑制控制海马网活动方面的贡献。这些毕业生和研究生的经历导致了三项第一作者初级研究和八项共同撰写/审查的出版物在同行评议的期刊上发表。为了寻找机会将自己的突触生理学经验融入更广泛的行为层面研究，奥尔廷斯基博士与宾夕法尼亚大学医学院的克里斯·皮尔斯博士进行了一系列会面。鉴于可卡因滥用研究中突触功能分析的相对不足，并意识到这是建立一条具有重大潜在影响的独立突触生理学研究体系的机会，奥尔丁斯基博士加入了皮尔斯博士的实验室，担任研究助理。在被任命后的短短时间里，奥尔丁斯基博士组装了一套脑片电生理记录的装置，并利用它(在皮尔斯博士的指导下)获得了大量数据，扩展了皮尔斯实验室最近的一系列发现。环境：宾夕法尼亚大学、附属地方中心和附近大学的基础成瘾研究社区非常突出，为咨询和合作创造了大量专业知识。在宾夕法尼亚大学充满活力的神经科学界内，由不同部门组织的会议和研讨会上有大量的科学互动机会。此外，几个校园办公室为博士后水平的生物医学教师制定了职业和研究技能培训计划。所有这些都为成功的研究和职业发展建立了广泛的资源丰富的环境。研究：在本申请中提出的研究建立在大量证据的基础上，即伏隔核中的多巴胺和谷氨酸系统之间的相互作用调节药物寻找和吸毒行为的重要方面。例如，包括皮尔斯博士在内的几个实验室已经证明，激活伏隔核中的D1样多巴胺受体，在一定程度上通过增加含有GluA1亚单位的AMPA受体向该脑区突触的运输，恢复了可卡因的寻求。然而，与整个大脑的神经元可塑性的研究相反，NMDA受体在可卡因诱导的神经元可塑性中的参与还没有得到系统的评估。我的初步结果表明，与对照组动物相比，在可卡因经历的动物身上，短暂刺激D1样多巴胺受体(如在可卡因自我给药期间发生的)，在伏隔核壳中的中等刺神经元上引起由NMDA受体介导的电流的持久增加。已知NMDA受体参与恢复对可卡因的依赖。因此，我们认为伏隔核壳中的D1样受体对NMDA受体功能的调节对持续的可卡因寻找行为有重要作用。职业发展计划：提出了一系列结合课程作业、会议和研讨会出席、正式研究报告、动手培训和教学的活动。正式课程将包括一门生物统计学研究生的统计方法和数据分析课程和一门神经精神药理学研究生课程。每年将参加两次研究会议：神经科学学会的年度会议，药物依赖问题学院的会议，或关于兴奋性突触和大脑功能的戈登会议。每年建议在不同的地点进行两到三次正式的研究报告。将通过参加讲习班和研讨会获得生物医学职业技能培训。在拟议的研究过程中将获得实践培训。教学计划包括几门研究生级别的神经科学课程的讲座。职业发展计划的总体目标是促进行为药理学和神经生理学方法在拟议研究中的整合，并为成瘾研究中成功的独立职业生涯奠定必要的技能基础。

DESCRIPTION (provided by applicant): The current application for a five-year mentored Career Development Award (K01) submitted by Dr. Pavel Ortinski proposes acquisition of training in the behavioral pharmacology of drug abuse. Specifically, the requested training will examine the role of D1-like dopamine and NMDA receptor interaction in an animal model of cocaine self-administration and reinstatement of cocaine seeking. This training will facilitate the development of an independent research program focused on the analysis of synaptic mechanisms of cocaine seeking and relapse. The research will be conducted at the University of Pennsylvania under the mentorship of Dr. Chris Pierce. Candidate: Dr. Ortinski has demonstrated a record of research projects implementing the analyses of synaptic function and physiology in several experimental models. His Ph.D. thesis, completed at Georgetown University under the mentorship of Dr. Stefano Vicini, used electrophysiological patch-clamp recordings in cell cultures and brain slices to investigate the regulation of inhibitory synaptic strength by GABA-A receptor subtypes. Dr. Ortinski then completed a two-year postdoctoral fellowship with Dr. Douglas Coulter at the Children's Hospital of Philadelphia/University of Pennsylvania where he evaluated contributions of the astrocytic glutamate/glutamine metabolic pathway to inhibitory control over hippocampal network activity in epilepsy. These graduate and post-graduate experiences resulted in three first-author primary research and eight co-authored/review publications in peer-reviewed journals. In search of opportunities to integrate his synaptic physiology experience into a broader context of behavioral-level studies, Dr. Ortinski had a series of meetings with Dr. Chris Pierce at the UPenn School of Medicine. Seeing a relative shortage of synaptic function analyses in cocaine abuse research and realizing this represented a chance to establish an independent line of synaptic physiology research with significant potential impact, Dr. Ortinski joined Dr. Pierce's laboratory as a Research Associate. In the short time since his appointment, Dr. Ortinski has assembled a set-up for electrophysiological recordings from brain slices and used it to acquire (with Dr. Pierce's guidance) a considerable amount of data extending a series of recent findings from the Pierce lab. Environment: The basic addiction research community at UPenn, affiliated local centers, and nearby universities is very prominent and creates a reservoir of expertise for consultation and collaboration. There are ample opportunities for scientific interactions at meetings and seminars organized by various departments within UPenn's vibrant neuroscience community. Additionally, several campus offices developed programs in career and research skill training for biomedical faculty at the postdoctoral level. All of these establish an extensive resource-rich environment for successful research and career development. Research: The research proposed in this application builds upon substantial evidence that interactions between dopamine and glutamate systems in the nucleus accumbens mediate important aspects of drug-seeking and drug-taking behaviors. For example, several laboratories, including that of Dr. Pierce, have demonstrated that activation of D1-like dopamine receptors in the nucleus accumbens reinstates cocaine seeking, in part, by increasing the trafficking of GluA1 subunit-containing AMPA receptors to synapses in this brain region. In contrast to studies of neuronal plasticity throughout the brain, however, the involvement of NMDA receptors in cocaine-induced neuronal plasticity has not been assessed systematically. My preliminary results show that, relative to control animals, transient stimulation of D1-like dopamine receptors (as occurs during cocaine self-administration) in cocaine-experienced animals elicits a long-lasting increase of currents mediated by NMDA receptors in the medium spiny neurons of the nucleus accumbens shell. NMDA receptors are known to be involved in reinstatement of cocaine-seeking. Thus, it is proposed that regulation of NMDA receptor function by D1-like receptors in the nucleus accumbens shell contributes significantly to persistent cocaine-seeking behaviors. Career development plan: A range of activities combining coursework, conference and workshop attendance, formal research presentations, hands-on training and teaching is proposed. Formal coursework will consist of a Statistical Methods and Data Analysis for Biostatistics graduate students course and a graduate Neuropsychopharmacology course. Two research conferences a year will be attended: an annual meeting of the Society for Neuroscience and either a meeting of the College of Problems on Drug Dependence or a Gordon conference on Excitatory Synapses and Brain Function. Two to three formal presentations of research yearly are proposed at various venues. Biomedical career skills training will be acquired through attendance of workshops and seminars. Hands-on training will be acquired in the course of the proposed research. Teaching plans include lectures in several graduate-level neuroscience courses. The overall goal of the career development plan is to facilitate the integration of behavioral pharmacology and neuronal physiology approaches in the proposed research and to establish a foundation of skills necessary for a successful independent career in addiction research.