ABSTRACT
[125I]17alpha-hydroxy-20alpha-yohimban-16beta-(N-4-p6 hydroxyphenethyl)carboxamide or [125I]rauwolscine-OHPC, a new radioiodinated probe derived from rauwolscine was synthesized and its binding characteristics investigated on sections of the mouse caudate putamen. [125I]rauwolscine-OHPC binding was saturable and revealed interaction with a single class of binding sites (KD= 0.171 nM, Bmax = 3082 pCi/mg of tissue). The kinetically derived affinity was in close agreement with the affinity evaluated by saturation experiments: k(-1)/k(+1)(0.0403 min(-1)/114 10(6) M(-1) min(-1))=0.35 nM. Competition studies revealed interaction with one single class of binding sites for each of the twelve compounds tested. The rank of potency suggested an interaction with alpha2 adrenoceptors (atipamezole > or = RX 821002 > yohimbine > (-)epinephrine). Moreover, the good affinity of [125I] rauwolscine-OHPC binding sites for spiroxatrine, yohimbine, WB 4101, the relatively good affinity for prazosin (Ki =37.4 nM) and the affinity ratio prazosin/oxymetazoline (37.4/43.4=0.86) were consistent with an alpha2C selective labelling of [125I]rauwolscine-OHPC. The distribution of [125I]rauwolscine-OHPC binding sites in mouse brain was characterized by autoradiography. The density of binding sites was high in the islands of Calleja, accumbens nucleus, caudate putamen and olfactory tubercles, moderate in the hippocampus, amygdala and anterodorsal nucleus of the thalamus. These findings demonstrated that [125I]rauwolscine-OHPC is a useful radioiodinated probe to label alpha2C adrenoceptors in mouse brain.
Subject(s)
Brain/metabolism , Receptors, Adrenergic, alpha-2/metabolism , Yohimbine/analogs & derivatives , Animals , Autoradiography , Binding Sites , Binding, Competitive , Caudate Nucleus/metabolism , Epinephrine/pharmacology , Iodine Radioisotopes , Kinetics , Male , Mice , Mice, Inbred Strains , Molecular Structure , Organ Specificity , Oxymetazoline/pharmacology , Prazosin/pharmacology , Putamen/metabolism , Receptors, Adrenergic, alpha-2/analysis , Yohimbine/chemical synthesis , Yohimbine/pharmacokinetics , Yohimbine/pharmacologyABSTRACT
OBJECTIVES: This study was conducted to compare microleakage of two new dentin bonding agents on freshly extracted teeth, cryopreserved teeth, or teeth stored in water containing 0.5% chloramine at 4 degrees C. METHODS: Rectangular Class V cavity preparations were made on the buccal and the lingual surface of wisdom teeth. They were filled with either Scotchbond Multi-Purpose and Z100 (3M Dental Products) or with Gluma 2000 and Pekafill (Bayer Dental). After thermocycling, silver staining penetration was evaluated under a light microscope. SEM examination and EDX analysis were performed to evaluate the microleakage pattern. The results were analyzed by the use of a two-way analysis of variance. RESULTS: Cryopreservation for 13 wk or 12 d refrigeration did not produce changes in the amount of microleakage. However, 48 d or longer of refrigeration increased microleakage. There was no correlation between changes in microleakage and storage time. Specimens prepared with both dentin bonding agents exhibited the same microleakage values and the same microleakage pattern. SIGNIFICANCE: Refrigeration at 4 degrees C in 0.5% chloramine for 48 d or longer may cause an increase in microleakage. Cryopreservation for 13 wk or short-term refrigeration did not affect the microleakage.
