STUDIES ON THE EFFECTS OF GLUCOSE INVITRO AND OF THE GLYCEMIC STATE INVIVO ON THE BINDING CHARACTERISTICS OF MU, DELTA-OPIATE AND KAPPA-OPIATE RECEPTORS IN MOUSE-BRAIN

The effect of glucose on the binding characteristics of opiate receptor subtypes was investigated in brain membranes from normoglycaemic lean Aston (C57BL/6J) mice using [H-3][D-Ala2,MePhe4,Gly5-ol]enkephalin (DAMGO), [H-3][D-Pen2,D-Pen5]enkephalin (DPDPE) and [H-3]U69,593 as selective ligands for mu, delta and kappa opiate receptors respectively. The equilibrium dissociation constants (K(d)) and maximal binding capacities (B(max)) of [H-3]DAMGO and [H-3]DPDPE were unaltered by 20mM glucose in vitro. Similarly, [H-3]U69,593 binding was not modified by increasing the concentration of glucose from 0 to 20mM (P between 0.10 and 0.05), or by the presence of 20mM fructose and of 20mM 3-0-me-glucose, a non-metabolisable sugar, in the incubation medium. The nonselective opiate ligand, [H-3]diprenorphine, bound with similar affinity and binding capacity to brain membranes prepared from control and streptozotocin-diabetic Swiss (CD1) mice. The addition of 20mM glucose or of 20mM fructose in vitro induced no changes in their binding parameters. The affinity and binding capacity of [H-3]U69,593 to STZ-diabetic Swiss mouse brain membranes was not significantly different to that of normoglycaemic controls; 20mM glucose in vitro had no effect on ligand binding to kappa sites in STZ-diabetic mouse brain membranes. We conclude that glucose does not interact directly with the opiate receptor to modify it in such as way as could explain the altered sensitivity to different opioid agonists seen in obese and hyperglycaemic animal models in vivo.