Use of slide-mounted frozen sections to study the autoradiographic localization, characterization and quantification of luteinizing hormone-releasing hormone receptors in the rat pituitary.
Slide-mounted frozen sections were used for the autoradiographic localization, characterization and quantification of LHRH receptors. The radioligand was the iodinated LHRH agonist [D-Ser(TBU)6, des-Gly-NH2(10)]LHRH ethylamide (LHRH-A). Specificity studies demonstrated that specific labeling was concentrated in the anterior pituitary lobe and that the tracer could be displaced with either LHRH or LHRH-A. Scatchard analysis of saturation curves gave one-site interactions with a high affinity (Kd = 0.48 +/- 0.1 nM) for the iodinated LHRH-A. In order to evaluate the effects of different treatments on the LHRH receptor concentration, the effects of sex steroids on the density of radioautographic reaction were evaluated using an image analyzer. The results obtained were similar to those previously reported with classical binding assays. Thus, this simple technique appears suitable for the study of localization and characterization of LHRH receptors in different experimental conditions.
Badr
M
M
MRC Group in Molecular Endocrinology, Le Centre Hospitalier de l'Université Laval, Ste-Foy, Quebec, Canada.
Pelletier
G
G
D000818:Animals; D001345:Autoradiography; D013196:Dihydrotestosterone / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D005629:Frozen Sections; D066298:In Vitro Techniques; D010052:Ovariectomy; D010903:Pituitary Gland, Anterior / Q000032:analysis* / Q000187:drug effects; D011374:Progesterone / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D011966:Receptors, LHRH / Q000032:analysis* / Q000187:drug effects
D011966:Receptors, LHRH; D013196:Dihydrotestosterone; D011374:Progesterone; D004958:Estradiol