Ascorbate injury and EDTA (or manganese) protection of D2-dopamine receptors

Ascorbic acid, EDTA, and Mn2+ modulated neuroleptic binding to membrane receptors but not to solubilized receptors. Preincubation with ascorbic acid at 22 degrees C produced a profound time-dependent decline in [3H]spiperone binding to membrane receptors (prepared in Tris-ascorbate buffer). Ascorbate had minimal effect if introduced simultaneously with [3H]spiperone in the incubation medium, and receptor binding was performed and assayed at 0 degrees C. Binding to the solubilized receptor was not ascorbate sensitive. Both EDTA and Mn2+ blocked the ascorbate effect on membrane receptors but were without effect on solubilized receptors. The preparation and incubation buffers used for [3H]spiperone binding studies should include EDTA if ascorbate is present in the buffer.