INTERACTION OF CONCANAVALIN-A AND WHEAT-GERM-AGGLUTININ WITH HELIX ACETYLCHOLINE-RECEPTORS

The effects of lectins concanavalin A (Con A) and wheat germ agglutinin (WGA), were studied on acetylcholine (ACh) responses of physically isolated internally dialyzed Helix aspersa neurons using the concentration clamp method and on the binding of [H-3]alpha-bungarotoxin to the cluster of neurons. Con A and WGA have different simple sugar specificity and produced different actions on ACh-evoked Cl conductance responses, which were antagonized by Con A (5 mug/ml) but were not altered by WGA. Con A depressed ACh responses when applied extracellularly while it had no effect on ACh responses of the same neuron when added to the intracellular solution, thus indicating that Con A specific glycoproteins are exposed on the surface of the neuron. The studies of the effect of Con A on the properties of the ACh binding site (receptor) have demonstrated, that (1) the onset of desensitization of ACh responses of the dialyzed neurons, determined from the decay of ACh-current from peak to plateau in the continued presence of agonist and best fitted by a double exponential function, was accelerated by Con A; (2) Con A depressed the maximal ACh induced current in a dose response relationship and altered the Hill coefficients; (3) Con A depressed the binding of [H-3]alpha-bungarotoxin to the cluster of neurons. These results indicate that Con A receptors on the surface of the neuronal membrane play a regulatory role in the ACh-receptor system and suggest that binding of lectin molecules to their receptors leads to inhibition of binding of ACh to ACh-receptors and to acceleration of the kinetics of desensitization of ACh receptors. All the effects of Con A, that is, on the peak amplitude, desensitization, dose-response relationship of ACh induced current and binding of [H-3]alpha-bungarotoxin, could be recovered by D-mannose, a competitive inhibitor of Con A binding to its receptor. The results also demonstrate, that studies of the effects of lectins on dialyzed snail neurons may provide useful information about distribution, properties and functional role of the carbohydrates exposed on the neuronal surface.