Flumazenil on Multiple Stress and Withdrawal Anxiety

描述（由申请人提供）：我们实验室的工作表明，反复的压力和戒断会导致戒断引起的焦虑对短暂暴露于慢性乙醇的敏感性，进一步支持Ballenger和Post（1978）的“点燃”假说。一个主要的发现是，氟马西尼，苯二氮卓类（BZD）拮抗剂，最大限度地减少焦虑的敏感性后，反复戒断协议。基于氟马西尼的这些数据，推测对GABAA受体具有负面作用的内源性物质负责由焦虑的重复应激致敏诱导的焦虑致敏。为了支持这一假设，给予两剂DMCM（一种BZD反向激动剂）而不是戒断导致最终戒断期间的焦虑敏感化。本调查的目的是测试具体的假设，以解释这些发现。具体目标1将确定是否反复应激致敏的戒断诱导的焦虑将拮抗氟马西尼，以及是否这种治疗的结果在mRNA表达增加地西泮结合抑制剂（DBI），内源性BZD-反向激动剂，允许识别的大脑部位DBI的适应性变化可能有助于戒断诱导的焦虑。具体目标2将测试重复应激或暴露于BZD反向激动剂是否会持续诱导戒断诱导焦虑的致敏作用，随后再次暴露于慢性乙醇，如重复戒断所发生的那样。此外，我们将确定在多次停药期间暴露于压力或BZD反向激动剂是否会延长持续时间，在此期间，随后再次暴露于慢性乙醇将继续导致焦虑敏感化。具体目标3将确定在具体目标1中确定的脑区域中微量注射氟马西尼是否会阻断多次应激后戒断诱导的焦虑的敏化，以及在选定部位中微量注射BZD反向激动剂是否会敏化戒断诱导的焦虑。具体目标4将测试GABA（A）和BZD结合是否受到重复应激和单次戒断的影响，以及对BZD反向激动剂的反应性是否因重复应激而增加。这将通过实验来补充，以确定是否对氟马西尼敏感的内源性化合物随着反复停药而增加。定义病理适应机制（S）的基础上负责“点燃”的撤退引起的焦虑与反复的压力可能会提供更深入的了解酒精滥用，并提供新的治疗途径。

DESCRIPTION (provided by applicant): Work in our laboratory has demonstrated that repeated stresses and withdrawals leads to sensitization of the withdrawal-induced anxiety to a brief exposure to chronic ethanol, in further support of the Ballenger and Post (1978)"kindling" hypothesis. A major finding is that flumazenil, a benzodiazepine (BZD) antagonist, minimizes the sensitization of anxiety following the repeated withdrawal protocol. Based upon these data with flumazenil, it is presumed that an endogenous substance having a negative effect on GABAA receptors is responsible for the sensitization of anxiety induced by the repeated stress sensitization of anxiety. In support of this hypothesis, two doses of DMCM, a BZD-inverse agonist, given instead of withdrawal results in sensitization of anxiety during a final withdrawal. The purpose of the present investigation is to test specific hypotheses to account for these findings. Specific Aim 1 will determine whether repeated stress sensitization of withdrawal-induced anxiety will be antagonized by flumazenil and whether this treatment results in an increase in mRNA expression for diazepam binding inhibitor (DBI), an endogenous BZD-inverse agonist, allowing identification of brain sites where adaptive change in DBI could be contributing to withdrawal-induced anxiety. Specific Aim 2 will test whether the repeated stresses or exposure to a BZD-inverse agonist will persist in inducing sensitization of withdrawal-induce anxiety upon later re-exposure to chronic ethanol, as occurs with repeated withdrawals. Additionally, we will determine if exposure to stress or a BZD-inverse agonist during multiple withdrawals will extend the duration during which later re-exposure to chronic ethanol will continue result in sensitization of anxiety. Specific Aim 3 will determine if microinjection of flumazenil into brain regions identified in Specific Aim 1 will block sensitization of withdrawal-induced anxiety following multiple stresses and whether microinjection of a BZD-inverse agonist into the selected sites will sensitize withdrawal-induced anxiety. Specific Aim 4 will test whether GABA(A) and BZD binding is affected by repeated stresses and a single withdrawal and whether responsiveness to a BZD-inverse agonist is increased by the repeated stresses. This will be complemented by experiments to identify whether an endogenous compound sensitive to flumazenil increases with repeated withdrawals. Defining the basis of the pathological adaptive mechanism(s) responsible for "kindling" of withdrawal-induced anxiety associated with repeated stresses may provide greater insight into alcohol abuse and provide new avenues for treatment.