Journal article

BINDING OF [H-3] AMPA TO NON-CHAOTROPE, NON-DETERGENT TREATED RAT SYNAPTIC-MEMBRANES - CHARACTERISTICS AND LACK OF EFFECT OF BARBITURATES

RC MORGAN, LD MERCER, M CINCOTTA, PM BEART

NEUROCHEMISTRY INTERNATIONAL | PERGAMON-ELSEVIER SCIENCE LTD | Published : 1991

Abstract

The binding of [(3)H]?-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) to synaptic membranes of rat brain was characterized in the absence of detergents and chaotropes. Optimal conditions for binding were a centrifugation assay employing a well washed, frozen-thawed synaptic membrane preparation and a 40 min incubation at 4 degrees C in 50 mM Tris-HC1/100 mM KC1, pH 7.4, buffer. Under these conditions, saturation experiments showed [(3)H]AMPA bound to high and low affinity sites (approx. K(d)s 150 nM and 15 ?M, respectively). Both components were also detectable when binding was terminated by filtration, but B(max) values were 20 times lower than found by centrifugation. Kainic aci..

View full abstract