Table_1_Functional Genomic Analysis of Amphetamine Sensitivity in Drosophila.XLSX

Abuse of psychostimulants, including amphetamines (AMPHs), is a major public health problem with profound psychiatric, medical, and psychosocial complications. The actions of these drugs at the dopamine transporter (DAT) play a critical role in their therapeutic efficacy as well as their liability for abuse and dependence. To date, however, the mechanisms that mediate these actions are not well-understood, and therapeutic interventions for AMPH abuse have been limited. Drug exposure can induce broad changes in gene expression that can contribute to neuroplasticity and effect long-lasting changes in neuronal function. Identifying genes and gene pathways perturbed by drug exposure is essential to our understanding of the molecular basis of drug addiction. In this study, we used Drosophila as a model to examine AMPH-induced transcriptional changes that are DAT-dependent, as those would be the most relevant to the stimulatory effects of the drug. Using this approach, we found genes involved in the control of mRNA translation to be significantly upregulated in response to AMPH in a DAT-dependent manner. To further prioritize genes for validation, we explored functional convergence between these genes and genes we identified in a genome-wide association study of AMPH sensitivity using the Drosophila Genetic Reference Panel. We validated a number of these genes by showing that they act specifically in dopamine neurons to mediate the behavioral effects of AMPH. Taken together, our data establish Drosophila as a powerful model that enables the integration of behavioral, genomic and transcriptomic data, followed by rapid gene validation, to investigate the molecular underpinnings of psychostimulant action.

包括苯丙胺类兴奋剂（amphetamines, AMPHs）在内的精神兴奋剂滥用，是一类可引发严重精神、躯体及社会心理并发症的重大公共卫生问题。此类药物通过作用于多巴胺转运体（dopamine transporter, DAT），其治疗功效与成瘾依赖潜质均与此密切相关。然而截至目前，介导此类作用的分子机制尚未得到充分阐释，针对苯丙胺类兴奋剂滥用的治疗干预手段仍十分有限。药物暴露可诱导基因表达发生广泛改变，进而促成神经可塑性变化，并对神经元功能产生长期影响。明确药物暴露所扰动的基因及基因通路，是理解药物成瘾分子基础的核心关键。本研究以果蝇（Drosophila）作为模式生物，探究苯丙胺类兴奋剂诱导的、依赖多巴胺转运体的转录变化——此类变化与药物的兴奋作用最为相关。通过该研究策略，我们发现参与调控mRNA翻译的基因，以依赖多巴胺转运体的方式在苯丙胺类兴奋剂处理后显著上调。为进一步筛选可供验证的候选基因，我们对上述基因与此前借助果蝇遗传参考品系（Drosophila Genetic Reference Panel）开展的苯丙胺类兴奋剂敏感性全基因组关联研究（genome-wide association study）中所鉴定的基因，进行了功能趋同性分析。我们通过实验验证了多个候选基因，证实其特异性作用于多巴胺能神经元，介导苯丙胺类兴奋剂引发的行为效应。综上，本研究证实果蝇是一款强大的研究模型，可整合行为学、基因组学与转录组学数据，并快速完成基因验证，用于探究精神兴奋剂作用的分子机制。