CELLULAR/BIOCHEMICAL NEUROPEPTIDE ADAPTATIONS ASSOCIATED WITH OPIATE TOLERANCE

阿片类药物耐受性和依赖性的潜在机制仍然是 被阐明。 对于这类现象已经提出了几种理论。 然而，最近的研究发现，内源性抗阿片肽（AOPs），如 神经肽FF（NPFF）和活性磷酸化阿片样物质微克 受体状态表明参与一个组合模型来解释 阿片类药物耐受性和依赖性的发展。 本项目 试图研究NPFF在调节结合参数中的相互作用 （受体密度和亲和力）和介导信号转导 SH-SY 5 Y细胞中阿片样物质受体系统的途径， 对吗啡耐受 细胞将通过暴露于 吗啡硫酸盐在1微克M浓度两天， 先前通过测量cAMP积累来显示。 匀浆结合 将检查NPFF和吗啡暴露后的研究。 此外，本发明还提供了一种方法， 慢性NPFF暴露在存在或不存在 吗啡耐受状态对cAMP积累、蛋白激酶C 活性和内源性磷酸化活性。 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耐受过程中。 的 这些研究的结果将有助于阐明 NPFF通过其减弱吗啡耐受性的发展。 此外，两名少数民族研究生参加了 拟议的项目将提高他们在生物医学的职业兴趣 研究，同时提供完善的实验室和实验研究 skills.

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.