PERIPHERAL BENZODIAZEPINE RECEPTOR CHARACTERIZATION IN MONKEYS WITH SIB

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 外周苯二氮卓类受体(PBR)存在于大脑和血小板中，已被证明在各种焦虑症中发挥重要作用。我们研究PBR的目标是确定被归类为SIB的猴子与对照组之间以及进入我们的安定治疗方案的自伤猴子之间基线受体密度的潜在差异。血液样本已经从所有SIB和对照动物中收集，并使用Gavish等人描述的技术分离了血小板。(1996年)。然而，在分析这些样本之前，我们一直在测试一个单独的冰冻血小板小球池，以进行分析优化试验。将颗粒均质，制备洗涤的膜部分，并使用选择性放射性配基(3上标H)PK-11195在饱和分析中测试所得到的组织样品。到目前为止，我们已经使用3-60 nm(3上标H)PK-11195的范围进行了几次分析，以及一次使用固定放射性配基浓度(10 NM)以及1-50 nm的非标记RO5-4864的“冷”饱和分析。用EBDA-Ligand软件(Elsevier-BIOSOFT)进行数据分析。我们继续努力优化检测方案，因为我们是第一个在任何猕猴物种中表征PBR结合的实验室。一旦分析程序充分制定，以产生可重现的动力学数据，我们将继续分析来自SIB和对照猴子的血小板样本。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Peripheral benzodiazepine receptors (PBR), which are present in the brain as well as in blood platelets, have previously been shown to play an important role in various anxiety disorders. Our goal for investigating PBR is to determine potential differences in baseline receptor density between monkeys classified as SIB versus controls, as well as in self-wounding monkeys that enter our diazepam treatment protocol. Blood samples have been collected from all of the SIB and control animals and the platelets isolated using a technique described in Gavish et al. (1996). However, before these samples are analyzed, we have been testing a separate pool of frozen platelet pellets for assay optimization trials. Pellets were homogenized, washed membrane fractions were prepared, and the resulting tissue samples were tested in a saturation analysis using the selective radioligand (3 superscript H) PK-11195. Thus far we have run several assays used a range of 3-60 nM (3 superscript H)PK-11195, as well as one "cold" saturation analysis used a fixed radioligand concentration (10 nM) along with 1-50 nM of non-labeled Ro 5-4864. Data were analyzed using EBDA-LIGAND software (Elsevier- BIOSOFT). We continue to work to optimize the assay protocol, as we are the first laboratory to characterize PBR binding in any macaque species. Once the assay procedures have been worked out adequately to yield reproducible kinetic data, we will proceed to analyze the platelet samples from the SIB and control monkeys.