PERIPHERAL BENZODIAZEPINE RECEPTOR CHARACTERIZATION IN MONKEYS WITH SIB

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 外周苯二氮卓受体（PBR），存在于大脑以及血小板中，先前已被证明在各种焦虑症中发挥重要作用。 我们研究PBR的目标是确定分类为SIB与对照组的猴子之间基线受体密度的潜在差异，以及进入我们的地西泮治疗方案的自伤猴子。 从所有SIB和对照动物中采集血样，并使用Gavish等人（1996）中描述的技术分离血小板。 然而，在分析这些样本之前，我们一直在测试一个单独的冷冻血小板沉淀池，用于分析优化试验。 将沉淀均质化，制备洗涤的膜组分，并使用选择性放射性配体（3上标H）PK-11195在饱和分析中测试所得组织样品。 到目前为止，我们已经使用3-60 nM（3上标H）PK-11195进行了几次测定，以及使用固定放射性配体浓度（10 nM）沿着1-50 nM未标记Ro 5-4864进行了一次“冷”饱和分析。 使用EBDA-LIGAND软件（Elsevier-BIOSOFT）分析数据。 我们将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。 一旦充分制定了试验程序以产生可重现的动力学数据，我们将继续分析SIB和对照猴的血小板样本。

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.