title: Changes in [(3)H]glibenclamide binding to mouse forebrain membranes during morphine tolerance.
abstract: The characteristics of specific binding of the ATP-sensitive K(+) (K(ATP)) channel blocker [3H]glibenclamide to forebrain membranes (P(2) fraction, 4 degrees C) obtained from morphine-naive and -tolerant mice were evaluated. Morphine tolerance was induced by osmotic minipumps that released 45 mg/kg/day of morphine subcutaneously for 6 days. This treatment enhanced the antinociceptive ED(50) of morphine without changing its E(max). In morphine-naive animals, (1) both the association and the dissociation of [3H]glibenclamide were biphasic; (2) [3H]glibenclamide was displaced by other sulfonylureas (order of potency: glibenclamide>glipizide&z.Gt;tolbutamide) with pseudo-Hill coefficients lower than unity and biphasic Hofstee plots; and (3) Scatchard plots of saturation experiments were curvilinear, showed a Hill coefficient of 0.81+/-0.04 and suggested the presence of two binding sites with a K(D) of 0.13 and 3.17 nM and a B(max) of 12.30 and 84.47 fmol/mg protein, respectively. By contrast, in membranes obtained from morphine-tolerant animals, (1) the Scatchard plots showed only one population of binding sites with a K(D) of 0.87 nM and a B(max) of 77.99 fmol/mg protein, and the Hill coefficient was very close to unity (0.96+/-0.1); (2) competition experiments (using glibenclamide as displacer) showed a pseudo-Hill coefficient of 0.99+/-0.04; and (3) dissociation experiments showed only one phase of dissociation. These results suggest that [3H]glibenclamide binds to two different sites in membranes obtained from morphine-naive animals, but to only one site in morphine-tolerant animals. Consequently, it seems that morphine tolerance in mice involves adaptive changes in K(ATP) channels.
authors.affiliation: Departamento de Farmacología e Instituto de Neurociencias, Facultad de Medicina, Universidad de Granada, Avda. Madrid 11, E-18012, Granada, Spain.
authors.forename: L G
authors.initials: LG
authors.lastname: González
authors.forename: E
authors.initials: E
authors.lastname: Portillo
authors.forename: E
authors.initials: E
authors.lastname: Del Pozo
authors.forename: J M
authors.initials: JM
authors.lastname: Baeyens
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D004361:Drug Tolerance; D005260:Female; D005905:Glyburide / Q000378:metabolism*; D006358:Hot Temperature; D020128:Inhibitory Concentration 50; D051379:Mice; D009020:Morphine / Q000494:pharmacology*; D010147:Pain Measurement; D015221:Potassium Channels / Q000378:metabolism; D016548:Prosencephalon / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D013997:Time Factors
chemical_list: D015221:Potassium Channels; D009020:Morphine; D000255:Adenosine Triphosphate; D005905:Glyburide