The role of lateral orbitofrontal cortex astrocytes in alcohol drinking

外侧眶额皮质星形胶质细胞在饮酒中的作用

基本信息

批准号：
10823447
负责人：
Abigail Riley Blasczyk
金额：
$ 4.77万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-19 至 2026-03-18
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10823447
关键词：
Action Potentials Acute Affect Agonist Air Alcohol consumption Alcohol dependence Alcohols Astrocytes Bilateral Brain region Calcium Calcium Signaling Categories Cell membrane Cells Chronic Complement Consumption Data Dependence Development Diagnosis Dopamine D1 Receptor Drug Addiction Electrophysiology (science)Ethanol Female Fiber Future Glycine Glycine Receptors Goals Health Heavy Drinking Helping Behavior Home Impairment Individual Infusion procedures Laboratories Lateral Learning Manuscripts Measures Membrane Potentials Modeling Monitor Mus Neurobiology Neurons Operative Surgical Procedures Opsin PMCA1 protein Persons Phase Photometry Physiology Play Positioning Attribute Potassium Channel Preparation Process Productivity Quinine Recreation Reporter Resistance Rest Role Slice Strychnine Substance Use Disorder Techniques Testing Therapeutic Intervention Training United States Viral Virus Water Withdrawal aged alcohol effect alcohol exposure alcohol research alcohol use disorder career career development dopamine D5 receptor drinking experimental study glycine transporter hippocampal pyramidal neuron in vivo inhibitor insight male negative affect neuron loss neuronal excitability novel optogenetics overexpression patch clamp prevent redshift research study response responsible research conduct social

项目摘要

PROJECT SUMMARY Alcohol use disorder (AUD) is characterized by the progression from recreational drinking to uncontrollable and excessive consumption resulting in a myriad of social and neurobiological complications. The mechanisms underlying the dependence-induced escalation in drinking are not completely understood. However, a key brain region disrupted in individuals with AUD is the orbitofrontal cortex (OFC). Studies from the Woodward laboratory show that acute ethanol inhibits action potential firing of lateral orbitofrontal (lOFC) cortex pyramidal neurons. This occurs via an astrocyte-dependent process involving activation of astrocytic D1/D5 dopamine receptors and the release of glycine via reversal of the GlyT1 glycine transporter. Following chronic intermittent exposure (CIE) to alcohol, lOFC neurons become hyperexcitable and are tolerant to acute ethanol. However, the effects of CIE exposure on lOFC astrocytes and how this affects lOFC neuronal excitability are not completely understood. The overarching hypothesis of this proposal is that CIE exposure impairs lOFC astrocyte function that contributes to hyperexcitability of lOFC pyramidal neurons and the resulting dependence-induced escalation in drinking. This hypothesis will be tested with two complementary aims. Aim 1 will test the hypothesis that CIE-induced increases in lOFC neuronal excitability and loss of acute ethanol inhibition involves astrocytic calcium signaling. To test this, male and female C57BL/6J mice will receive an intra-OFC infusion of an astrocyte-selective AAV encoding either a plasma membrane calcium ATPase (PMCA) or a reporter construct (tdTomato). Following repeated cycles of CIE exposure, slice electrophysiology will be used to measure current-evoked spiking of lOFC neurons and the membrane potential of lOFC astrocytes. Training in viral infusion surgeries and astrocyte and neuron slice electrophysiology will be achieved under this aim. Aim 2 will test the hypothesis that expressing PMCA in lOFC astrocytes prevents the increases in drinking following CIE exposure. In the first study, male and female C57BL/6J mice expressing either PMCA or tdTomato localized in lOFC astrocytes will undergo baseline sessions of two-bottle choice ethanol drinking. Weekly sessions of CIE exposure will then be interleaved with test weeks of drinking. The second study will follow the same CIE paradigm with the absence of homecage drinking and will use mice expressing GCaMP6f in lOFC astrocytes and the red-shifted opsin ChrimsonR in lOFC neurons. Training in astrocyte fiber photometry and optogenetics will be achieved under this aim. The proposed research studies will be complemented by career development activities including manuscript preparation, data presentation, networking, and training in the responsible conduct of research. These studies will provide novel insight into the role of lOFC astrocytes in AUD and position me to pursue a productive career in alcohol research.

项目摘要酒精使用障碍（AUD）的特征是从休闲饮酒到无法控制和过度消费导致无数的社会和神经生物学并发症。机制尚不完全了解依赖性引起的饮酒升级。但是，关键的大脑 AUD个体中断的区域是Orbitrontal皮层（OFC）。伍德沃德实验室的研究表明急性乙醇抑制了侧轨道额（LOFC）皮质锥体神经元的作用电势触发。这是通过星形胶质细胞依赖性过程发生的，涉及星形胶质细胞D1/D5多巴胺受体的激活甘氨酸通过甘氨酸转运蛋白的逆转释放。慢性间歇性暴露（CIE）对于酒精而言，LOFC神经元可过度转变，并且对急性乙醇具有耐受性。但是，CIE的影响在LOFC星形胶质细胞上的暴露以及这如何影响LOFC神经元兴奋性。这该提案的总体假设是CIE暴露会损害LOFC星形胶质细胞功能，这有助于 LOFC锥体神经元和由此产生的依赖引起的饮酒升级的过度兴奋。这假设将通过两个互补目标进行检验。 AIM 1将检验CIE引起的增加的假设在LOFC神经元的兴奋性和急性乙醇抑制作用的丧失涉及星形胶质性钙信号传导。测试这是男性和雌性C57BL/6J小鼠，将收到ACROCYTE-SEXEXIDE AAV编码的INC内部输注质膜ATPase（PMCA）或报告基因构建体（TDTOMATO）。以下重复 CIE暴露的周期，切片电生理学将用于测量LOFC神经元的诱发电流尖峰以及LOFC星形胶质细胞的膜电位。病毒输注手术，星形胶质细胞和神经元的培训在此目标下将实现切片电生理学。 AIM 2将检验以下假设： LOFC星形胶质细胞可防止CIE暴露后饮酒的增加。在第一个研究中，男性和女性表达PMCA或TDTOMATO的C57BL/6J小鼠将在LOFC星形胶质细胞中进行基线会话两瓶选择乙醇饮用。然后，每周的CIE暴露会议将与测试周交织喝酒。第二项研究将遵循相同的CIE范式，没有归乡饮酒，将在LOFC星形胶质细胞中使用表达GCAMP6F的小鼠和LOFC神经元中的红移Opsin Chrimsonr。在此目标下将实现星形胶质细胞纤维光度法和光遗传学的培训。拟议的研究研究将由职业发展活动进行补充，包括手稿准备，数据在负责任的研究行为中的演示，网络和培训。这些研究将提供新颖深入了解LOFC星形胶质细胞在AUD中的作用，并将我从事酒精研究的富有成效的职业。