Coupling of I(1) imidazoline receptors to the cAMP pathway: studies with a highly selective ligand, benazoline.
Clonidine and benazoline are two structurally related imidazolines. Whereas clonidine binds both to alpha(2)-adrenoceptors (alpha(2)R) and to I(1) imidazoline receptors (I(1)R), benazoline showed a high selectivity for imidazoline receptors. Although the alpha(2)R are negatively coupled to adenylate cyclase, no effect on cAMP level by activation of I(1)R has been reported so far. We therefore aimed to compare the effects of clonidine and benazoline on forskolin-stimulated cAMP levels in cell lines expressing either I(1)R only (PC12 cells), alpha(2)R only (HT29 cells), or I(1)R and alpha(2)R together (NG10815 cells). Clonidine proved able to decrease the forskolin-stimulated cAMP level in the cells expressing alpha(2)R and this effect could be blocked by rauwolscine. In contrast, in cells lacking these adrenoceptors, clonidine had no effect. On the other hand, benazoline and other I(1) receptor-selective imidazolines decreased forskolin-stimulated cAMP level in the cells expressing I(1)R, in a rauwolscine- and pertussis toxin-insensitive manner. These effects were antagonized by clonidine. According to these results, we demonstrated that 1) alpha(2)R and I(1)R are definitely different entities because they are expressed independently in different cell lines; 2) alpha(2)R and I(1)R are both implicated in the cAMP pathway in cells (one is sensitive to pertussis toxin and the other is not); and 3) I(1)R might be coupled to more then one transduction pathway. These new data will be essential to further understand the physiological implications of the I(1)R and the functional interactions between I(1) receptors and alpha(2)-adrenoceptors.
Greney
H
H
Laboratoire de Neurobiologie et Pharmacologie Cardiovasculaire, Faculté de Medecine, Université Louis Pasteur, Strasbourg, France.
Ronde
P
P
Magnier
C
C
Maranca
F
F
Rascente
C
C
Quaglia
W
W
Giannella
M
M
Pigini
M
M
Brasili
L
L
Lugnier
C
C
Bousquet
P
P
Dontenwill
M
M
D037361:Adenylate Cyclase Toxin; D000316:Adrenergic alpha-Agonists / Q000494:pharmacology; D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D000818:Animals; D003000:Clonidine / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D019073:HT29 Cells; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D054755:Imidazoline Receptors; D008024:Ligands; D016716:PC12 Cells; D037342:Pertussis Toxin; D051381:Rats; D018341:Receptors, Adrenergic, alpha-2 / Q000378:metabolism; D011955:Receptors, Drug / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D015016:Yohimbine / Q000494:pharmacology
D037361:Adenylate Cyclase Toxin; D000316:Adrenergic alpha-Agonists; D000317:Adrenergic alpha-Antagonists; D007093:Imidazoles; D054755:Imidazoline Receptors; D008024:Ligands; D018341:Receptors, Adrenergic, alpha-2; D011955:Receptors, Drug; D010566:Virulence Factors, Bordetella; D015016:Yohimbine; D000242:Cyclic AMP; D037342:Pertussis Toxin; D003000:Clonidine; C010242:metizoline