CELLULAR/BIOCHEMICAL NEUROPEPTIDE ADAPTATIONS ASSOCIATED WITH OPIATE TOLERANCE

阿片类药物耐受和依赖的潜在机制(S)仍是 将被澄清。对于这种现象，人们提出了几种理论。 然而，最近发现的内源性抗阿片肽(AOPS)类似 神经肽FF(NPFF)和活性磷酸化阿片微克 受体状态提示参与了一种组合模型来解释 阿片类药物耐受性和依赖性的发展。目前的项目 尝试检查NPFF在调节绑定参数时的交互作用 (受体密度和亲和力)以及在介导信号转导中的作用 阿片微克再捕获系统在SH-SY5Y细胞中的途径 对吗啡耐受。细胞将通过暴露于 在浓度为1微克M的硫酸吗啡中持续两天，这有 以前通过测量营地积累显示了这一点。匀浆结合 将对NPFF和吗啡暴露后的研究进行审查。此外, 存在或不存在慢性NPFF暴露的调制效应 吗啡耐受状态对cAMP积聚、蛋白激酶C的影响 活性和内源性磷酸化活性也将被执行。 本项目旨在确定NPFF的这种相互作用 在受体和亚细胞水平的阿片类药物耐受期间。这个 这些研究的结果将有助于描述这种机制。 NPFF通过其抑制吗啡耐受的发展。 此外，两名少数族裔研究生参加了 拟议的项目将增强他们对生物医学的职业兴趣 研究，同时提供健全的实验室和实验研究 技能。

The underlying mechanism(s) of opiate tolerance and dependence remains to be elucidated. Several theories have been postulated for such phenomena. However, recent findings of endogenous anti-opiate peptides (AOPs) like neuropeptide FF (NPFF), and an active phosphorylated opioid microgram receptor state suggest the involvement of a combined model to explain the development of opioid tolerance and dependence. The present project attempts to examine NPFF interaction in modulating the binding parameters (receptor density and affinity) and in mediating the signal transduction pathway of the opioid microgram recaptor system in the SH-SY5Y cells that are morphine tolerant. Cells will be made tolerant by being exposed to morphine sulfate at a 1 microgram M concentration for two days, which has been previously shown by measuring cAMP accumulation. Homogenate binding studies after NPFF and morphine exposure will be examined. In addition, the modulatory effects of chronic NPFF exposure in the presence or absence of a morphine tolerant state on cAMP accumulation, protein kinase C activity and endogenous phosphorylation activity will also be performed. The present project is designed to determine such interactions of NPFF during opiate tolerance at both the receptor and subcellular levels. The results generated from these studies will help to delineate the mechanisms by which NPFF attenuates the development of morphine tolerance. Furthermore, participation by two minority graduate students in the proposed project will enhance their career interests in biomedical research, while providing sound laboratory and experimental research skills.