PROTRH DERIVED PEPTIDES DURING OPIATE WITHDRAWAL

这项研究的长期目标是更好地了解生理 机制和化学介质负责阿片戒断。 我们已经观察到，一个独特的神经元群体位于 中脑导水管周围灰质（PAG）的腹外侧区显示 促甲状腺激素原释放显著增加（约500%） 激素（促甲状腺激素释放激素原）基因表达在阿片类药物戒断。 以来 该区域的兴奋引起了一种静止的和低反应的模式， 行为，假设腹外侧区的proTRH神经元 PAG被激活作为对过度活跃的 阿片类药物戒断状态 进一步假设，一个或多个 proTRH衍生肽可介导这些反应。 我们提出 为了阐明这个独特的群体的解剖连接性， 在腹外侧PAG中的阿片反应性proTRH神经元， 共聚焦激光显微镜和电子显微镜作为一种方法来识别 大脑中参与阿片类药物戒断的特定通路 反应，并确定这些神经元是如何整合到 对吗啡过度活跃状态作出反应的控制系统 戒断 此外，我们将确定细胞特异性 这些神经元对阿片类药物戒断的反应是由于 这些细胞上存在阿片受体，如果CREB或 CREB相关蛋白的上调介导这些反应。 因为这些神经元所处的位置 参与疼痛的调节，proTRH基因的增加 持续性深部伤害性刺激后PAG的活动 将确定麻醉动物。 促甲状腺激素释放激素原的作用 肽在阿片戒断和抗伤害作用将进一步 通过使用腺病毒抑制proTRH基因表达来研究 含有反义proTRH转基因的载体，立体定向 以及它们对行为和自主神经的影响 量化戒断期间的反应。 同时，将确定 PAG中的proTRH衍生肽在阿片类药物作用期间增加 通过色谱和电泳技术提取，以及 基于这种分析，将合成肽并注射到 来测量它对戒断的影响 应答 随着海洛因成瘾继续在共同的药物 滥用，从这个建议产生的数据可能有临床 在设计新的治疗方法方面的意义 成瘾和戒断综合征。

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.