Imidazolinic radioligands for the identification of hamster adipocyte alpha 2-adrenoceptors

Eur J Pharmacol. 1989 Nov 21;171(2-3):145-57. doi: 10.1016/0014-2999(89)90103-9.

Abstract

Imidazolinic radioligands ([3H]UK 14304, [3H]idazoxan and [3H]RX 821002) were used for the identification of alpha 2-adrenoceptors on hamster fat cell membranes since there are limitations to the use of [3H]yohimbine and [3H]clonidine, which suggest alpha 2-adrenoceptor heterogeneity. Biological assays (lipolysis measurements) were performed on isolated fat cells and binding studies were carried out on fat cell membranes. The imidazolinic derivative, UK 14304, was a full agonist as compared to clonidine. Idazoxan and RX 821002 (2-(2-methoxy-1,4-benzodioxan-2yl)-2-imidazoline), a recently developed alpha 2-antagonist, were more potent alpha 2-antagonists than yohimbine in this fat cell model. [3H]UK 14304 was the most suitable agent for the quantification of the 'high-affinity state' alpha 2-adrenoceptors in binding studies since it did not exhibit the sensitivity to the composition of the buffer shown by [3H]clonidine. Although it is a potent alpha 2-antagonist, [3H]idazoxan had major limitations for use in the identification of alpha 2-adrenoceptors in this cell model since it also bound to 'non-adrenaline displaceable' binding sites which were revealed when imidazolinic derivatives (phentolamine) were used instead of adrenaline to determine the non-specific binding. We demonstrated that [3H]RX 821002 was a more suitable radioligand than [3H]yohimbine for labelling hamster fat cell alpha 2-adrenoceptors (KD = 1.0 +/- 0.1 nM, Bmax = 776 +/- 60 fmol/mg protein). Moreover, since it exhibited low affinity for 'imidazoline-preferring sites', it represents a valuable ligand even in tissues possessing such binding sites. We suggest that [3H]RX 821002 can be used to identify alpha 2-adrenoceptors in various tissues when these sites cannot be labelled with [3H]yohimbine.

MeSH terms

  • Adipose Tissue / drug effects
  • Adipose Tissue / metabolism*
  • Adrenergic alpha-Agonists / pharmacology
  • Adrenergic alpha-Antagonists / pharmacology
  • Animals
  • Brimonidine Tartrate
  • Clonidine / pharmacology
  • Cricetinae
  • Dioxanes / pharmacology
  • Idazoxan
  • Imidazoles / metabolism*
  • In Vitro Techniques
  • Kinetics
  • Lipid Metabolism
  • Lipolysis
  • Male
  • Mesocricetus
  • Quinoxalines / pharmacology
  • Radioligand Assay
  • Receptors, Adrenergic, alpha / metabolism*
  • Yohimbine / pharmacology

Substances

  • Adrenergic alpha-Agonists
  • Adrenergic alpha-Antagonists
  • Dioxanes
  • Imidazoles
  • Quinoxalines
  • Receptors, Adrenergic, alpha
  • Yohimbine
  • Brimonidine Tartrate
  • 2-methoxyidazoxan
  • Clonidine
  • Idazoxan