Synaptic changes in the medial prefrontal cortex in the development of compulsive alcohol drinking

强迫性饮酒发展过程中内侧前额叶皮层的突触变化

• 批准号: 10367079
• 负责人: SVEN KROENER
• 金额: $ 36.25万
• 依托单位: UNIVERSITY OF TEXAS DALLAS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-15 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10367079
• 关键词: Acute; Affect; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcohols; Amygdaloid structure; Animal Model; Animals; Automobile Driving; Back; Behavior; C57BL/6 Mouse; CREB1 gene; Cells; Chloride Channels; Chronic; Cognitive; Cognitive deficits; Cues; Data; Dendritic Spines; Dependence; Development; Electrophysiology (science); Ethanol; Genetic Recombination; Glutamate Receptor; Glutamates; Goals; Immunohistochemistry; Impaired cognition; Impulsive Behavior; Interneurons; Label; Light; Lithium Chloride; Medial; Modeling; Morphology; Motivation; Mus; N-Methyl-D-Aspartate Receptors; Neurons; Nucleus Accumbens; Output; Pathway interactions; Pharmaceutical Preparations; Play; Population; Prefrontal Cortex; Pyramidal Cells; Relapse; Reporter; Resistance; Resolution; Rodent; Role; Self Administration; Self-Direction; Synapses; Synaptic plasticity; Taste aversion; Testing; Time; Vertebral column; Withdrawal; Work; activity marker; alcohol availability; alcohol exposure; alcohol seeking behavior; cognitive change; cognitive function; combinatorial; drinking; drug of abuse; drug seeking behavior; executive function; experimental study; flexibility; hippocampal pyramidal neuron; improved; incentive salience; learning extinction; neuroadaptation; novel; optogenetics; patch clamp; postsynaptic; presynaptic; problem drinker; receptor expression; receptor function; receptor upregulation; relapse prediction; response; synaptic function; vapor; voltage clamp

SUMMARY: The ability to inhibit drinking is a significant challenge for recovering alcoholics, especially in the presence of alcohol- associated cues. Repeated alcohol exposure induces neuroadaptations that persist beyond acute withdrawal, and which increase alcohol’s incentive salience, leading to escalation of alcohol intake and aversion-resistant alcohol seeking. Alcohol use also causes deficits in cognitive functions associated with the medial prefrontal cortex (mPFC), which further fuel compulsive drinking and relapse. In rodents, alcohol seeking activates specialized networks within the ventral (infralimbic, IL) and prelimbic (PL) regions of the mPFC, which play largely opposite roles in the control of relapse behavior. While activation of the PL drives reinstatement, neurons in the IL facilitate extinction learning and inhibit drug-seeking through their projections to the Nucleus Accumbens shell, as well as the basolateral amygdala (BLA). However, the synaptic mechanisms that drive maladaptive plasticity in these circuits during the transition from controlled to compulsive alcohol- seeking remain largely unclear. The experiments in this application will provide a better understanding of network-specific mechanisms through which chronic alcohol exposure and withdrawal affect executive cognitive functions of the mPFC and diminish inhibitory control over goal-directed behavior. In Aim 1 we use patch-clamp electrophysiology and optogenetic stimulation to determine changes in long-range glutamatergic inputs from the BLA onto identified projection neurons in the IL and PL following extended access to alcohol, as well as under postdependent conditions. We will use Targeted Recombination in Active Populations (TRAP2) with Fos2A- iCreER mice to express channelrhodopsin selectively in those BLA afferents to the mPFC that are activated during withdrawal, and we will determine how alterations in these inputs develop over time (from goal-directed to compulsive alcohol- seeking). We will perform voltage-clamp recordings from retrogradely-labeled neurons that project back to the BLA, and we will determine alcohol-induced changes in postsynaptic glutamate receptor function and presynaptic release following reinstatement. Experiments in Aim 2 will use combinational retrograde Cre delivery and a Cre-dependent reporter to label the same IL and PL projection neurons for high-resolution morphometric analyses of spines and glutamate receptor expression in order to compare changes specifically in those neurons that contribute to relapse behavior (visualized via co-labeling for the activity marker phospho-CREB) and those that do not (pCREB-negative cells). In Aim 3 we will again use TRAP2 mice to test whether optogenetic manipulations of specific ensembles in the mPFC, or of inputs from the BLA to the mPFC (each again TRAPed during withdrawal) can reverse alcohol-induced cognitive deficits and reduce drug- seeking. Taken together, these studies will provide important novel information about alcohol-induced synaptic changes in networks of the IL that contribute to cognitive flexibility and cue-induced reinstatement.

概括： 抑制饮酒的能力是恢复酗酒者的重大挑战，尤其是在酗酒的情况下 - 相关提示。反复的酒精暴露会引起神经适应的持续超出急性感谢和 增加酒精的激励显着性，从而导致酒精摄入量升级和寻求抗厌食症的酒精。 使用还会导致与介质前额叶皮层（MPFC）相关的认知功能的防御能力，这进一步燃料 强迫性饮酒和救济。在啮齿类 MPFC的IL）和前比（PL）区域，它们在控制继电器行为中起着很大相反的作用。尽管 PL的激活驱动器恢复原状，IL中的神经元促进扩展学习并抑制通过药物寻求药物 他们的项目投向了伏隔核的壳，以及基体杏仁核（BLA）。但是，突触 从受控酒精到强迫性酒精的过渡期间，这些电路中驱动不良适应性可塑性的机制 寻求仍然不清楚。本应用程序中的实验将为特定网络提供更好的了解 慢性酒精暴露和戒断的机制影响了MPFC的执行认知功能 减少对目标指导行为的抑制作用。 在AIM 1中，我们使用斑板钳电生理学和光遗传学刺激来确定远距离的变化 谷氨酸能在BLA上输入到IL中鉴定的投影神经元，并在扩展到酒精后PL， 以及在依赖后条件下。我们将使用fos2a-中的活动人群（TRAP2）中使用靶向重组 iCreer小鼠在撤回期间被激活的那些BLA传入中选择性地表达ChannelRhopoptin 我们将确定这些输入的变化如何随着时间的流逝而发展（从目标定向到强迫性酒精 - 寻求）。我们将从逆行标记的神经元中执行电压钳记录，这些神经元将其投射回BLA，并且 我们将确定酒精引起的突触后谷氨酸受体功能和突触前释放的变化 复职。 AIM 2中的实验将使用逆行CRE递送和CRE依赖的记者来标记 用于刺和谷氨酸受体的高分辨率形态分析的同一IL和PL投影神经元 表达以比较有助于继电行为的神经元中的变化（通过 为活性标记磷酸化 - 磷酸化）和没有的（pCREB阴性细胞）的共同标记。在目标3中，我们将再次 使用TRAP2小鼠测试MPFC中特定合奏的光学遗传操作还是来自BLA的输入 到MPFC（在撤离期间再次出现）可以逆转酒精引起的认知缺陷并减少药物 - 寻求。 综上所述，这些研究将提供有关酒精引起的突触变化的重要新信息 IL的网络有助于认知灵活性和提示引起的恢复。