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： 抑制饮酒的能力对于戒酒者来说是一个重大挑战，尤其是在酒精存在的情况下。 反复接触酒精会诱发神经适应，这种适应在急性戒断后仍持续存在。 增加酒精的刺激显着性，导致酒精摄入量和厌恶酒精寻求的增加。 使用还会导致与内侧前额叶皮层 (mPFC) 相关的认知功能缺陷，从而进一步加剧 在啮齿类动物中，酗酒会激活腹侧（边缘下、 mPFC 的 IL）和前边缘（PL）区域在控制复发行为中发挥着相反的作用。 PL 的激活驱动恢复，IL 中的神经元通过促进消退学习并抑制药物寻求 它们投射到伏隔核壳以及基底外侧杏仁核 (BLA)。 在从控制性酒精到强迫性酒精的转变过程中，驱动这些回路的适应不良可塑性的机制 该应用程序中的实验将提供对特定网络的更好理解。 慢性酒精暴露和戒断影响 mPFC 执行认知功能的机制 减少对目标导向行为的抑制控制。 在目标 1 中，我们使用膜片钳电生理学和光遗传学刺激来确定长程变化 在长期接触酒精后，来自 BLA 的谷氨酸输入到 IL 和 PL 中已识别的投射神经元上， 以及在后依赖性条件下，我们将使用 Fos2A- 的活性群体靶向重组 (TRAP2)。 iCreER 小鼠在戒断期间激活的 mPFC 的 BLA 传入神经中选择性表达通道视紫红质， 我们将确定这些输入随着时间的推移如何发生变化（从目标导向到强迫性饮酒） 我们将从投射回 BLA 的逆行标记神经元进行电压钳记录，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 目标 2 中的实验将使用组合逆行 Cre 传递和 Cre 依赖性报告基因来标记。 相同的 IL 和 PL 投射神经元用于脊柱和谷氨酸受体的高分辨率形态测量分析 表达，以便比较那些导致复发行为的神经元中的具体变化（通过可视化 共同标记活性标记磷酸化 CREB）和那些不标记磷酸化 CREB ​​的细胞（pCREB ​​阴性细胞）。 使用 TRAP2 小鼠来测试是否对 mPFC 中的特定整体或来自 BLA 的输入进行光遗传学操作 mPFC（在戒断期间再次陷入困境）可以逆转酒精引起的认知缺陷并减少药物- 寻求。 总而言之，这些研究将提供关于酒精诱导的突触变化的重要新信息。 IL 网络有助于认知灵活性和线索诱导的恢复。