title: Functional receptors in Xenopus oocytes for U-37883A, a novel ATP-sensitive K+ channel blocker: comparison with rat insulinoma cells.
abstract: Follicle-enclosed Xenopus oocytes were used to describe the ATP-sensitive K+ (KATP) channel-blocking properties of U-37883A (4-morpholinecarboximidine-N-1-adamantyl-N'-cyclohexyl), in comparison with glibenclamide. In follicular oocytes, the KATP channel opener P1060 (30 microM), a pinacidil analog, activated a large outward K+ current that was blocked by glibenclamide (IC50 = 0.33 microM) and U-37883A (IC50 = 0.26 microM). P1060 activation was inhibited by both U-37883A and glibenclamide in a noncompetitive manner. U-37883A also blocked the KATP channel activation by cAMP (300 microM) and adenosine (10 microM). Single-channel studies on isolated follicular cells showed that U-37883A (10 microM) reduced the open probability of the KATP channel by 76%, without significantly modifying the single-channel current amplitude. Receptor binding studies with [3H]U-37883 in membranes from follicle-enclosed oocytes demonstrated a single class of low affinity binding sites, with a Kd of 450 nM and a Bmax of 17 pmol/mg of protein. Studies with analogs of U-37883A showed that U-52090A inhibited KATP current and displaced [3H]U-37883 from its binding site with similar potencies. In contrast, U-42069D neither inhibited KATP current nor competed with [3H]U-37883 binding. In RINm5F cells (an insulinoma cell line), U-37883A, unlike glibenclamide, failed to inhibit KATP current. Furthermore, there was no significant specific binding of [3H]U-37883 in RINm5F cell membranes, which displayed high levels of specific binding of [3H]glibenclamide. These data demonstrate the presence of a receptor for U-37883A-type guanidines that controls the activity of the endogenous KATP channels in follicle-enclosed oocytes. The available data collectively suggest that U-37883A is a more selective blocker of the follicular KATP channel, which is very similar to that in smooth muscle, than of the pancreatic beta cell KATP channel.
authors.affiliation: Institut de Pharmacologie Moleculaire et Cellulaire, Centre Nationale de la Recherche Scientifique, Valbonne, France.
authors.forename: E
authors.initials: E
authors.lastname: Guillemare
authors.forename: E
authors.initials: E
authors.lastname: Honore
authors.forename: J
authors.initials: J
authors.lastname: De Weille
authors.forename: M
authors.initials: M
authors.lastname: Fosset
authors.forename: M
authors.initials: M
authors.lastname: Lazdunski
authors.forename: K
authors.initials: K
authors.lastname: Meisheri
mesh_terms: D000218:Adamantane / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D005260:Female; D005905:Glyburide / Q000494:pharmacology; D007340:Insulinoma / Q000473:pathology; D009025:Morpholines / Q000378:metabolism / Q000494:pharmacology*; D009865:Oocytes / Q000378:metabolism*; D015221:Potassium Channels / Q000187:drug effects*; D051381:Rats; D014407:Tumor Cells, Cultured; D014982:Xenopus laevis
chemical_list: D009025:Morpholines; D015221:Potassium Channels; C082658:U 37883A; D000255:Adenosine Triphosphate; D000218:Adamantane; D005905:Glyburide