Effects of Opiates on Cellular Mechanisms

描述(由申请人提供)： 本申请是一项已资助了5年的高级科学家奖的延续申请。申请者至少75%的时间将用于研究，其余的时间将用于教学和服务中与研究相关的活动。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职员工。申请者将继续在“NIDA之友”中担任领导职务，这是一个他帮助发展起来的组织，在科学家和非科学家中倡导科学。该机构的日常运作将由一家受雇公司负责，以免影响申请者本人在本申请中所描述的研究项目。将在下一个五年周期内开展的研究将是申请者获得NIDA奖励的资助研究的继续。他是R01基金“脑啡肽、神经药理学和滥用潜力”的首席研究员，也是项目项目基金中“大麻类急性和慢性影响中的细胞系统”项目的首席研究员。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信号转导中涉及到这两类滥用药物的急性影响和耐受性的步骤。我们已经证明，蛋白激酶A(PKA)和蛋白激酶C(PKC)都能逆转小鼠对吗啡的8倍镇痛耐受、低温耐受和Straub尾部改变。我们还表明，需要同时抑制PKA和PKC，才能逆转吗啡颗粒植入后3天发生的45倍耐受性。然而，在慢性吗啡暴露的18天中，对PKC和PKA抑制剂耐受逆转的抵抗力逐渐发展，这表明替代途径被激活。PKA的特异性抑制剂PKI-TID逆转了3天的吗啡耐受，并逆转了吗啡耐受小鼠脊髓中PKA水平的增加。本申请中描述的工作旨在继续阐明吗啡耐受逆转与大脑选定区域PKA或PKC活性变化之间是否存在相关性。我们的实验室还表明，PKA抑制剂还可以逆转Delta-9 THC产生的明显耐受性。目前，我们正在研究长期使用大麻类药物对脑区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、β-9THC与内源性大麻素、阿南达胺以及合成的更稳定的阿南达胺类似物之间的交叉耐受性。本申请中描述的工作将旨在阐明在使用Delta-9 THC慢性治疗后是否发生脑内PKA和/或脑内PKC的变化，以及使用稳定的阿南达胺类似物的慢性治疗是否会产生类似的影响。还将评估这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.