PROTRH DERIVED PEPTIDES DURING OPIATE WITHDRAWAL

这项研究的长期目标是更好地了解 阿片类药物戒断的机制和化学媒介。 我们观察到一组独特的神经元位于 中脑导水管周围灰质腹外侧区(PAG)显示 促甲状腺激素原释放显著增加(约500%) 阿片类药物戒断过程中激素(ProTRH)基因的表达。自.以来 这一区域的兴奋唤起了一种静止和低反应的模式 行为，假设腹外侧区的前TRH神经元 激活的PAG是对过度活跃的一种代偿反应 鸦片类药物戒断状态。还假设一个或多个 ProTRH衍生的多肽可能介导这些反应。我们建议 为了阐明这一独特种群的解剖连接性 阿片受体反应性前TRH神经元在PAG腹外侧区的应用 激光共聚焦显微镜和电子显微镜作为一种识别 大脑中参与阿片类药物戒断的特定通路 并确定这些神经元是如何整合到 对吗啡过度活跃状态做出反应的控制系统 戒烟。此外，我们将确定细胞是否具有特异性 这些神经元对阿片类药物戒断的反应是由于 这些细胞上阿片受体的存在，如果CREB或 CREB相关蛋白的上调介导了这些反应。 因为这些神经元所在的位置也可能 参与痛觉调制的前TRH基因增加 持续深度伤害性刺激后PAG的活动 麻醉的动物将被确定。PRTRH衍生的作用 多肽在阿片戒断和抗伤害作用中的作用将进一步 用腺病毒抑制前TRH基因表达的研究 含有反义原TRH转基因的载体，立体定位 PAG内注射及其对行为和自主神经的影响 戒断过程中的反应被量化。同时，将标识 阿片类药物作用下PAG内PRTRH衍生肽的增加 通过层析和电泳技术退出，以及 基于这一分析，多肽将被合成并注入到 大脑的不同区域，以测量其对戒断的影响 回应。由于海洛因成瘾继续在常见的毒品上 滥用，这项提案产生的数据可能具有临床意义 在设计新的治疗方法方面的意义 成瘾和戒断综合症。

The long term goal of this study is to better understand the physiologic mechanisms and chemical mediators responsible for opiate withdrawal. We have observed that a unique population of neurons located in the ventrolateral region of the midbrain periaqueductal gray (PAG) show a marked increase (approximately 500%) in pro-thyrotropin-releasing hormone (proTRH) gene expression during opiate withdrawal. Since excitation of this region evokes a quiescent and hyporeactive pattern of behavior, it is hypothesized that proTRH neurons in the ventrolateral PAG are activated as a compensatory response to the hyperactive state of opiate withdrawal. It is further hypothesized that one or more proTRH-derived peptides may mediate these responses. We propose to elucidate the anatomical connectivity of this unique population of opiate-responsive proTRH neurons in the ventrolateral PAG using confocal laser microscopy and electron microscopy as a way to identify specific pathways in the brain that are involved in opiate-withdrawal responses and determine how these neurons are integrated into the control system that responds to the hyperactive state of morphine withdrawal. In addition, we will determine whether the cell specific responses of these neurons to opiate withdrawal are due to the presence of opiate receptors on these cells and if CREB or upregulation of CREB-related proteins mediate these responses. Because these neurons are in a location where they could also be involved in the modulation of pain, an increase in proTRH gene activity in the PAG following deep noxious stimulation in continuously anesthetized animals will be determined. The role of proTRH-derived peptides in opiate withdrawal and antinociception will be further studied by inhibition of proTRH gene expression using adenovirus vectors containing an antisense proTRH transgene, stereotaxically injected into the PAG and their effect on behavioral and autonomic responses during withdrawal quantified. In parallel, will identify the proTRH-derived peptides in the PAG that increase during opiate withdrawal by chromatographic and electrophoretic techniques, and based on this analysis, peptides will be synthesized and injected into discrete regions of the brain to measure its effect on withdrawal responses. As heroin addiction continues to be on the common drugs of abuse, the data generated from this proposal could have clinical significance in the design of new approaches to the treatment of addiction and the withdrawal syndrome.