Role of Endocannabinoid Signaling in Alcoholism

描述（由申请人提供）： 酒精（EtOH）抑制兴奋性神经传递并增强抑制性神经传递，而由突触后神经元释放的AEA显示通过激活突触前神经元上的CB 1受体（CB 1 R）来抑制神经递质释放。本项目的目的是更好地了解花生四烯酸酰胺（AEA）和CB1Rs在乙醇对颗粒神经元、中间神经元和浦肯野细胞之间突触的神经传递作用中的作用。在我们的方法中，我们使用小脑混合神经元培养。这种制备提供了独特的可能性，即颗粒细胞和中间神经元与浦肯野细胞建立突触接触，就像它们在完整的小脑中所做的那样，并且已知使用谷氨酸或GABA作为神经递质。我们的研究将集中在AEA和CB1Rs在EtOH对自发和诱发的兴奋性和抑制性突触传递的作用中的作用。另外的实验还将使用来自CB1R敲除小鼠幼仔的细胞测试新的AEA受体是否参与EtOH对自发和诱发的突触传递的作用。在第一个具体的目标，我们将测试是否AEA和CB1Rs参与的行动EtOH的自发和诱发的兴奋性和抑制性突触传递。我们将在存在或不存在受体调节剂的情况下测量急性EtOH暴露细胞中的EPSC和IPSC振幅以及mEPSC和mlPSC频率和振幅。在具体目标2中，我们将测量慢性EtOH诱导的CB1R和FAAH免疫反应性的变化以及AEA和CB1R在慢性EtOH对IPSC和EPSC振幅以及mlPSC和mEPSC频率和振幅的影响中的作用。在目标3中，我们将研究CB1Rs的缺乏如何影响急性和慢性EtOH对IPSC和EPSC振幅以及mlPSC和mEPSC频率和振幅的影响。更好地了解这些机制将产生新的药物治疗与潜在的治疗酒精中毒。

DESCRIPTION (provided by applicant): Alcohol (EtOH) suppress the excitatory neurotransmission and potentiate the inhibitory neurotransmission whereas AEA released by postsynaptic neurons is shown to inhibit neurotransmitter release by activating CB1 receptors (CB1 R) at presynaptic neurons. The purpose of this project is to gain a better understanding of anandamide (AEA) and CB1Rs role in the action of EtOH on neurotransmission at synapses between granular neurons, interneuron, Purkinje cells. In our approach we use cerebellar mixed neurons in culture. This preparation offers the unique possibility that granular cells and interneurons establish synaptic contacts with Purkinje cells as they do in the intact cerebellum and are known to use glutamate or GABA as a neurotransmitter. Our researches will focus on the role of AEA and CB1Rs in the action of EtOH on the spontaneous and evoked excitatory and inhibitory synaptic transmission. Additional experiments will also test whether or not novel AEA receptors are involved in the action of EtOH on the spontaneous and evoked synaptic transmission using cells from CB1R knockout mice pups. In the first specific aim, we will test whether or not AEA and CB1Rs are involved in the action of EtOH on the spontaneous and evoked excitatory and inhibitory synaptic transmission. We will measure the EPSC and IPSC amplitude and mEPSC and mlPSC frequency and amplitude in acute EtOH exposed cells in the presence or absence of receptor modulators. In specific aim 2, we will measure the chronic EtOH-induced changes in CB1R and FAAH immunoreactivity and the role of AEA and CB1Rs in the effects of chronic EtOH on IPSC and EPSC amplitudes and mlPSC and mEPSC frequency and amplitude. In aim 3, we will study how the lack of CB1Rs influences the effects of acute and chronic EtOH on IPSC and EPSC amplitudes and mlPSC and mEPSC frequency and amplitude. The better understanding of these mechanisms will yield new drug therapies with potential in treatment of alcoholism.