Identification of a D1 dopamine receptor, not linked to adenylate cyclase, on lactotroph cells.
1. We studied the lactotroph cells of the rat by both in vivo and in vitro pharmacological techniques for the presence of D1-receptors. Both approaches revealed the presence of D2-receptor, stimulated by quinpirole (resulting in an inhibition of prolactin secretion) and blocked by domperidone. 2. Administration of fenoldopam, the most selective D1-receptor agonist currently available, resulted in a dose-dependent decrease of prolactin secretion in vivo (after pretreatment with alpha-methyl-p-tyrosine) and in vitro (cultured pituitary cells). This increase was dose-dependently blocked by the selective D1-receptor antagonist, SCH 23390, and although the effect of fenoldopam was less than that obtained by D2-receptor stimulation, these data suggest that a D1-receptor also controls prolactin secretion. 3. In order to detect the location of these dopamine receptors, autoradiographic studies were performed by use of [3H]-SCH 23390 and [3H]-spiperone as markers for D1- and D2-receptors, respectively. Specific binding sites for [3H]-SCH 23390 were demonstrated. Fenoldopam dose-dependently reduced [3H]-SCH 23390 binding, but had no effect on [3H]-spiperone binding. Immunocytochemical labelling of prolactin cells after incubation with [3H]-SCH 23390 revealed that the granulae and hence, D1 binding sites were present on the lactotroph cells. 4. Radioligand binding studies performed on membranes from anterior pituitary cells revealed the presence of the D2-receptor (54 fmol mg-1 protein) with a Kd of 0.58 nM for [3H]-spiperone, but failed to detect D1-receptors. 5. Finally, we studied the effect of dopamine and of fenoldopam on the adenosine 3':5'-cyclic monophosphate (cyclic AMP) content of anterior pituitary cells. Although cyclic AMP increased upon prostacyclin administration, indicating an intact adenylate cyclase system, fenoldopam failed to increase the cyclic AMP production. 6. It is tempting to speculate that fenoldopam reduces prolactin secretion through interaction with a non-cyclase-linked D1-receptor on the lactotroph cells.
Schoors
D F
DF
Department of Pharmacology, Medical School, Vrije Universiteit Brussel V.U.B., Belgium.
Vauquelin
G P
GP
De Vos
H
H
Smets
G
G
Velkeniers
B
B
Vanhaelst
L
L
Dupont
A G
AG
D000262:Adenylyl Cyclases / Q000032:analysis; D000818:Animals; D001345:Autoradiography; D001552:Benzazepines / Q000032:analysis; D002408:Catheters, Indwelling; D004305:Dose-Response Relationship, Drug; D005260:Female; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D010903:Pituitary Gland, Anterior / Q000737:chemistry*; D011388:Prolactin / Q000032:analysis / Q000378:metabolism*; D011869:Radioligand Assay; D051381:Rats; D011919:Rats, Inbred Strains; D011954:Receptors, Dopamine / Q000032:analysis*; D017447:Receptors, Dopamine D1; D013134:Spiperone / Q000032:analysis; D014018:Tissue Distribution
D001552:Benzazepines; D011954:Receptors, Dopamine; D017447:Receptors, Dopamine D1; D013134:Spiperone; D011388:Prolactin; D000262:Adenylyl Cyclases