Circuit and epigenetic mechanisms underlying incubation of methamphetamine craving

线索诱导的药物渴求在戒断后逐渐增加，这种现象被称为“潜伏期”。 对毒品的渴望在过去的十年中，机制研究主要集中在可卡因的孵化 渴望相反，很少有研究探讨了潜伏期的机制， 甲基苯丙胺（Methamphetamine）在这里，我建议研究电路和表观遗传机制， 潜伏期的甲基渴望，重点是背侧纹状体（DS）区域。选择这个大脑区域是基于 我的初步数据暗示了DS神经元活动的关键作用和DS组蛋白的潜在作用 脱乙酰酶5（HDAC 5，一种表观遗传相关酶）在甲基渴望的孵化中的作用。在电路层面，我将 结合神经元活动使用逆行追踪技术（CT B，霍乱毒素亚单位B 标记Fos以识别在甲基苯丙胺渴求的孵化期间激活的对DS的关键传入投射。 接下来，我将使用一种新的retro-DREADD（设计师受体仅由设计师药物激活）方法， 研究传入投射到DS的因果作用在孵化的甲基渴望。在表观遗传学的层面上，我会 联合收割机染色质免疫沉淀测序（ChIP-seq）与RNA测序（RNA-seq）相结合， DS中HDAC 5的下游基因靶点，其表达在长期停药（1个月）后发生变化 从甲安菲他明自我管理。我还将使用CRISPR-Cas9系统来研究DS HDAC 5的因果作用。 对冰毒的渴望我的提议将为冰毒渴求的潜在机制提供新的见解 和复发此外，在Shaham、Nestler和科万博士的指导下，该提案将提供 我有一个理想的培训环境，为我未来的独立职业生涯做好准备， 探索药物成瘾基本机制的学术机构。

Cue-induced drug craving progressively increases after withdrawal, a phenomenon termed ‘incubation of drug craving’. During the last decade, mechanistic studies have primarily focused on incubation of cocaine craving. In contrast, very few studies have examined the mechanisms underlying incubation of methamphetamine (Meth) craving. Here, I propose to study circuit and epigenetic mechanisms underlying incubation of Meth craving with a focus on the dorsal striatum (DS) region. The choice of this brain area is based on my preliminary data implicating a critical role of DS neuronal activity and a potential role of DS histone deacetylase 5 (HDAC5, an epigenetic-related enzyme) in incubation of Meth craving. At the circuit level, I will use a retrograde tracing technique (CTb, cholera toxin subunit B) in combination with the neuronal activity marker Fos to identify critical afferent projections to the DS that are activated during incubation of Meth craving. Next, I will use a novel retro-DREADD (Designer Receptors Exclusively Activated by Designer Drugs) method to study the causal role of afferent projections to the DS in incubation of Meth craving. At the epigenetic level, I will combine chromatin immunoprecipitation-sequencing (ChIP-seq) with RNA-sequencing (RNA-seq) to identify downstream gene targets of HDAC5 in DS whose expression is altered after prolonged withdrawal (1 month) from Meth self-administration. I will also use the CRISPR-Cas9 system to examine the causal role of DS HDAC5 in incubation of Meth craving. My proposal will provide new insights on the mechanisms underlying Meth craving and relapse. Additionally, under the mentorship of Drs. Shaham, Nestler, and Cowan the proposal will provide me with an ideal training environment to prepare me for a future independent career as a researcher in an academic setting who explores basic mechanisms of drug addiction.