Central amygdala input circuits control stress-induced anxiety after chronic ETOH

杏仁核中央输入电路控制慢性 ETOH 后压力引起的焦虑

• 批准号: 9303762
• 负责人: GEORGE R BREESE
• 金额: $ 39.65万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-15 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9303762
• 关键词: Abstinence; Address; Affect; Agonist; Alcoholism; Alcohols; Amygdaloid structure; Anxiety; Brain; Chronic; Clinical; Complement; Distant; Ethanol; Functional Magnetic Resonance Imaging; Glutamates; Halorhodopsins; Hypothalamic structure; Injectable; Investigation; Knowledge; Lateral; Link; Medial; Modeling; Neurons; Output; Oxytocin; Oxytocin Receptor; Pharmacology; Play; Rhodopsin; Role; Series; Site; Stress; Symptoms; Synapses; Testing; Thalamic structure; Therapeutic; Vasopressin Antagonist; Vasopressin Receptor; Vasopressins; alcohol abstinence; alcohol exposure; anxiety-like behavior; base; craving; expression vector; improved; innovation; midbrain central gray substance; negative affect; neural circuit; neuropathology; optogenetics; prevent; problem drinker; promoter; public health relevance; relating to nervous system; response; social; vector

DESCRIPTION (provided by applicant): During abstinence from alcohol, stress in alcoholics can result in negative affect and craving (1), a response which is accompanied by a change in brain functional magnetic resonance imaging (fMRI)-effects of stress not seen in social drinkers (2). To model the negative affect induced by stress during abstinence in alcoholics, an extended period of chronic intermittent alcohol (CIA) exposure was found to cause an enduring adaptation that sensitizes stress-induced negative affect during abstinence. These series of clinical and basic findings are consistent with the kindling/stress hypothesis of alcoholism. Even though the central amygdala (CeA) is known to play an important role in stress-induced negative affect after CIA exposure, unknown is the role input circuits to the CeA have in facilitating this stress-induced anxiety. In addition to CRF, other inputs that synapse on CeA neurons are glutamate (GLU) terminals from the basolateral amygdala (BLA), oxytocin (OXY) terminals from the hypothalamus, and vasopressin (VP ) terminals. Nonetheless, the influence these other inputs have on stress-induced anxiety after CIA exposure has not been identified. Likewise unidentified is whether CeA medial division (CEM) neurons that output terminals to the periaqueductal gray (PAG) and other sites are a critical component of the neural circuit that supports stress-induced anxiety associated with CIA. To guide addressing these unknowns, the hypothesis tested is that stress facilitates anxiety during abstinence from CIA exposure by influencing a neural circuit composed of terminal inputs to the lateral (CEL) and medial (CEM) divisions of the CeA that modulate CEM output. First, to permit optogenetic investigations of the potential role BLA terminals have in facilitating stress-induced anxiety after CIA exposure, an AAV5-eYFP vector with a CAMKII promoter containing the rhodopsin derivatives halorhodopsin (NpHR3.0) or channel- rhodopsin (ChR2) will be placed into the BLA to confirm localization of GLU terminals from the BLA to the lateral (CEL) and the medial (CEM) divisions of the CeA. Subsequently tested with optogenetics will be whether ChR2 activation or NpHR3.0 inhibition of BLA terminal release of GLU on CEL neurons will influence facilitation of stress-induced anxiety after CIA exposure. This latter optogenetic strategy will be complemented by determining if activation of CEL neurons with an OXY receptor agonist will block facilitation of stress-induced anxiety after CIA exposure. Upon completing investigations of CEL neural inputs, optogenetic inhibition or excitation of GLU-containing BLA terminals synapsing on CEM neurons will assess if the stress-induced anxiogenic action after CIA exposure is affected. Because VP-containing terminals synapse on CEM neurons to induce anxiety- like behavior, it is reasoned that activation of CEM neurons with VP may contribute to the anxiety induced by stress following CIA exposure. To confirm a direct involvement of CEM neurons in VP action, an AAV-eYFP vector with the NpHR3 expressed in CEM neurons will test if optogenetic inhibition of CEM neurons will alter both stress- as well as VP-induced anxiety after CIA exposure. To confirm that VP action contributes to stress-induced anxiety after CIA exposure, VP receptor subtypes will be pharmacologically antagonized in the CEM prior to stress to determine if the stress- induced anxiety after CIA exposure is prevented. Upon confirmation of CEM terminal presence in the PAG, involvement of these CEM terminals in the PAG in stress- and VP-induced anxiety observed after CIA exposure will be assessed by optogenetic inhibition or excitation of these CEM terminals- an approach to implicate both the CEM and the PAG in facilitation of the stress-induced anxiety after CIA exposure. Collectively, the proposed pharmacological and optogenetic strategies utilized can be expected to define whether involvement of specific neural inputs to the CEL and CEM, which are accompanied by CEM output to the PAG and other sites, form a circuit that influences facilitation of stress-induced negative affect after CIA exposure. This proposed circuit involving the CeA is felt to be associated with the neuropathology responsible for the facilitated negative affect observed to stress that facilitates craving in alcoholics durin abstinence. New knowledge concerning adaptations in this circuit after CIA exposure could possibly provide clues by which to minimize the barrier to developing new and improved therapeutics to treat the negative symptoms to stress observed in abstinent alcoholics.

描述（由申请人提供）：在戒酒期间，酗酒者的压力会导致负面影响和渴望（1），这种反应伴随着脑功能磁共振成像（fMRI）的变化-在社交饮酒者中看不到压力的影响（2）。为了模拟酗酒者在禁欲期间由压力引起的负面影响，发现长期慢性间歇性酒精（CIA）暴露会引起持久的适应，使禁欲期间压力引起的负面影响敏感化。这一系列的临床和基础研究结果与酒精中毒的点燃/应激假说相一致。即使中央杏仁核（CeA）是已知的CIA暴露后，在压力引起的负面影响中发挥重要作用，未知的是输入电路的作用CeA在促进这种压力引起的焦虑。除了CRF，在CeA神经元上突触的其他输入是来自基底外侧杏仁核（BLA）的谷氨酸（GLU）终末、来自下丘脑的催产素（OXY）终末和加压素（VP）终末。尽管如此，这些其他输入的影响CIA暴露后的压力引起的焦虑尚未确定。同样未确定的是，是否CeA内侧区（CEM）神经元输出终端的中脑导水管周围灰质（PAG）和其他网站是一个关键组成部分的神经回路，支持与CIA相关的压力诱导的焦虑。为了指导解决这些未知数，测试的假设是，压力促进焦虑从CIA曝光禁欲期间通过影响由终端输入到外侧（CEL）和内侧（CEM）部门的CeA调制CEM输出的神经回路。首先，为了允许光遗传学研究BLA末端在CIA暴露后促进应激诱导的焦虑中的潜在作用，将具有CAMKII启动子的AAV 5-eYFP载体（包含视紫红质衍生物盐视紫红质（NpHR3.0）或通道视紫红质（ChR 2））放入BLA中，以确认GLU末端从BLA定位到CeA的外侧（CEL）和内侧（CEM）分区。随后用光遗传学测试ChR 2激活或NpHR3.0抑制CEL神经元上GLU的BLA末端释放是否会影响CIA暴露后应激诱导的焦虑的易化。后一种光遗传学策略将通过确定用OXY受体激动剂激活CEL神经元是否会阻断CIA暴露后应激诱导的焦虑的促进来补充。在完成CEL神经输入的研究后，CEM神经元上突触的含GLU的BLA末端的光遗传学抑制或激发将评估CIA暴露后应激诱导的致焦虑作用是否受到影响。由于含VP的终末在CEM神经元上突触以诱导焦虑样行为，因此有理由认为VP对CEM神经元的激活可能有助于CIA暴露后由应激诱导的焦虑。为了证实CEM神经元直接参与VP作用，具有在CEM神经元中表达的NpHR 3的AAV-eYFP载体将测试CEM神经元的光遗传学抑制是否将改变CIA暴露后的应激以及VP诱导的焦虑。为了证实VP作用有助于CIA暴露后的应激诱导的焦虑，将在应激前在CEM中拮抗VP受体亚型，以确定CIA暴露后的应激诱导的焦虑是否被预防。在确认PAG中存在CEM末端后，将通过这些CEM末端的光遗传学抑制或兴奋来评估PAG中的这些CEM末端在CIA暴露后观察到的应激和VP诱导的焦虑中的参与-一种暗示CEM和PAG两者在CIA暴露后促进应激诱导的焦虑的方法。总的来说，所提出的药理学和光遗传学策略可以预期用于定义是否涉及到CEL和CEM的特定神经输入，其伴随着CEM输出到PAG和其他部位，形成影响CIA暴露后应激诱导的负面影响的促进的回路。这种涉及CeA的拟议回路被认为与神经病理学有关，该神经病理学负责观察到的对压力的易化负面影响，这种易化负面影响促进了酗酒者在戒酒期间的渴望。关于CIA暴露后该电路适应性的新知识可能提供线索，以最大限度地减少开发新的和改进的治疗方法来治疗禁欲酗酒者中观察到的压力阴性症状的障碍。