This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor 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.
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