Effects of Opiates on Cellular Mechanisms

描述（由申请人提供）： 这份申请是对一项已经资助了5年的高级科学家奖的延续申请。申请人至少75%的时间将用于研究，其余时间用于研究教学和服务相关活动。他还将继续担任NIDA资助的机构培训赠款的PI。他还将继续指导研究生、博士后研究员和年轻教师。申请人将继续在“NIDA之友”中担任领导角色，该组织是他帮助在科学家和非科学家中开展科学宣传的组织。该机构的日常运作将由一家受雇的公司处理，以免影响申请人在本申请中描述的自己的研究项目。在下一个五年周期内进行的研究将是申请人获得NIDA资助的研究的延续。他担任R 01赠款的主要研究员，题为“脑啡肽，神经药理学和滥用潜力”和一个项目，题为“细胞系统在大麻素的急性和慢性影响”在一个程序项目赠款。本申请中提出的研究是这些项目的组合。贯穿这些项目的一个共同主题是确定与这两类滥用药物的急性效应和耐受性有关的信号转导步骤。蛋白激酶A（PKA）和蛋白激酶C（PKC）均能逆转吗啡对镇痛、低温和Straub尾改变的8倍耐受。我们还表明，需要抑制PKA和PKC来逆转吗啡颗粒植入后3天发生的45倍耐受性。然而，耐受性逆转PKC和PKA抑制剂的发展逐步超过18天的慢性吗啡暴露，表明替代途径的激活。PKI-tide是PKA的特异性抑制剂，可逆转吗啡耐受3天的小鼠脊髓中PKA水平的升高。本申请中描述的工作旨在继续阐明吗啡耐受性的逆转与大脑选定区域中PKA或PKC活性的改变之间是否存在相关性。在我们的实验室中还显示，PKA抑制剂也逆转了由δ-9 THC产生的明显耐受性。目前，我们正在研究大麻素长期治疗对大脑区域PKA水平的影响。在其他研究中，我们已经证明了外源性大麻素、δ-9 THC和内源性大麻素、大麻素和合成的更稳定的大麻素类似物之间的交叉耐受性。本申请中描述的工作将针对阐明在用δ-9 THC长期治疗后是否发生脑PKA和/或脑中PKC的变化，以及用花生四烯酸的稳定类似物长期治疗是否发生类似的效果。还将评估两类大麻素之间的交叉耐受性。

DESCRIPTION (provided by applicant): This application is a request for the continuation of a senior scientist award that has been funded for 5 years. At least 75% of the applicant's time will be directed to research, and the rest to research related activities in teaching and service. He will also continue to be the PI of the NIDA funded institutional training grant. He also will continue to mentor graduate students, post-doctoral fellows and young faculty members. The applicant will continue to serve in a leadership role in "The Friends of NIDA", an organization that he helped to develop for science advocacy among scientists and non-scientists. The everyday operation of this organization will be handled by a hired firm so as not to detract from the applicant's own research projects described in this application. The research to be carried out during the next five year cycle will be a continuation of funded research the applicant has been awarded by NIDA. He serves as the principal investigator of an R01 grant entitled "Enkephalins, Neuropharmacology and Abuse Potential" and of a project entitled "Cellular systems in the acute and chronic effects of cannabinoids" in a program project grant. The research proposed in this application is a combination of those projects. A common theme that flows through these projects is to identify the steps in signal transduction that are involved in the acute effects and tolerance that develops to these two families of abused drugs. We have demonstrated that protein kinase A (PKA) and protein kinase C (PKC) both reverse 8 fold morphine tolerance to the analgesic, hypothermia and changes in Straub tail in mice. We have also shown that one needs to inhibit both PKA and PKC to reverse a 45-fold tolerance occurring 3-days after morphine pellet implantation. However, a resistance to tolerance reversal by PKC and PKA inhibitors develops progressively over 18 days of chronic morphine exposure, indicating the activation of alternative pathways. PKI-tide a specific inhibitor of PKA reversed 3-day morphine tolerance and reversed the increase in PKA levels found in the spinal cord of morphine tolerant mice. The work described in this application is designed to continue to elucidate whether a correlation exists between reversal of morphine tolerance and an alteration of PKA or PKC activity in selected areas of the brain. It has also been shown in our laboratories that PKA inhibitors also reverse the pronounced tolerance produced by delta-9 THC. At the present time, we are in the process of examining the effect of long term treatment with cannabinoids on levels of PKA in brain regions. In other studies we have demonstrated cross tolerance between the exogenous cannabinoid, delta-9 THC and the endogenous cannabinoid, anandamide and synthetic more stable analogs of anandamide. Work described in this application will be directed toward elucidating whether changes in brain PKA and/or PKC in brain occur after chronic treatment with delta-9 THC and whether similar effects occur with chronic treatment with stable analogs of anandamide. Cross tolerance between the two classes of cannabinoids will also be evaluated.