10,11-Dihydro-5-methyl-5H-dibenzo(a,d)cyclohepten-5,10-imine → Dizocilpine

Participants

Reaction Steps

[3H]MK-801 binding was performed using Triton X-100 washed synaptic plasma membranes (SPM) prepared from rat forebrain (30-45 day old, male Sprague-Dawley; Sasco, St. Charles, MO) as described previously [J. W. Thomas, W. F. Hood, J. B. Monahan, P. C. Contreras and T. L. O'Donohue, Brain Res., 442, 396-398 (1988)]. The assay was initiated by the addition of SPM (0.20-0.30 mg) to an incubation containing 2.0 nM [3H]MK-801 (15 Ci/mmole; New England Nuclear, Boston, MA) and various concentrations of the appropriate test compound in a total volume of 0.5 ml (all additions were made in 50mM Tris/acetate buffer, pH 7.4) and continued for 120 min at 25° C. The samples were then filtered through glass fiber filters (Schleicher and Schuell #32) which were pretreated with 0.05% (v/v) polyethylenimine. The filters were washed and the radioactivity quantitated by liquid scintillation spectrometry. Inhibition of [3H]MK-801 binding was measured as a decrease in specific basal binding (basal binding=2583 ±381 DPM) with nonspecific binding as the residual binding in the presence of 60 μM MK-801 (562 ±30 DPM). The IC50 values for the inhibition of [3H]MK-801 binding were determined using a logit-log analysis and are reported in Table III, below.

Provenance & Evidence