Characterization of 17-beta-estradiol-dependent and -independent somatostatin receptor subtypes in rat anterior pituitary.
Previous studies from this laboratory showed that treatment with 17-beta-estradiol (E2) caused an acquisition of inhibitory effect of somatostatin (SRIF) on prolactin release with an increased number of SRIF-binding sites in the rat anterior pituitary. The aim of this study was to characterize the E2-dependent SRIF receptor in comparison with the E2-independent one, which was expressed in ovariectomized rats. The following observations were obtained: 1) both of the E2-dependent and E2-independent SRIF receptors, measured with 125I-Tyr11-SRIF as a radiolabeled ligand, were enriched in the plasma membrane fraction of the cells, displaying a single class of binding site (E2-dependent: Kd, 32 pM, Bmax, 2.3 pmol/mg protein; E2-independent: Kd, 83 pM, Bmax, 0.26 pmol/mg protein). The ligand binding to both receptors was sensitive to monovalent and divalent cations, and GTP. 2) Among the SRIF analogs tested, the relative potencies of SRIF28 and its analog and cyclosomatostatin compared with SRIF were lower in the E2-dependent receptor than in the E2-independent one. 3) A cross-linking study with N-hydroxysuccinimidyl-4-azido-benzoate revealed that the molecular weight of the cross-linked E2-dependent receptor was approximately 94,000, whereas that of the E2-independent one was 82,000, irrespective of the presence of a reducing reagent. The molecular weight of SRIF receptor from normal male or female rat pituitary was similar to the E2-independent type. 4) Both types of the cross-linked SRIF receptors were solubilized by sucrose monolaurate, adsorbed to a wheat germ agglutinin-agarose column, and eluted with N-acetyl-glucosamine. 5) SRIF inhibited the forskolin-stimulated adenylate cyclase activity in the pituitary membranes from E2-treated rats, but it did not in the E2-depleted membranes. These results demonstrate that there are at least two subtypes of SRIF receptor in the rat anterior pituitary, one of which is exclusively expressed by the treatment with E2, and that these subtypes are distinct with respect to ligand binding specificity, molecular weight, and coupling to adenylate cyclase inhibition.
Kimura
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Department of Molecular Neurobiology, Tokyo Metropolitan Institute for Neuroscience, Japan.
Hayafuji
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C
Kimura
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N
D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000502:physiology; D000818:Animals; D001667:Binding, Competitive; D002412:Cations / Q000494:pharmacology; D002772:Cholera Toxin / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004958:Estradiol / Q000502:physiology*; D005260:Female; D008024:Ligands; D008297:Male; D008970:Molecular Weight; D010903:Pituitary Gland, Anterior / Q000502:physiology*; D051381:Rats; D011919:Rats, Inbred Strains; D017981:Receptors, Neurotransmitter / Q000145:classification / Q000502:physiology*; D017481:Receptors, Somatostatin; D013004:Somatostatin / Q000378:metabolism*; D013347:Subcellular Fractions / Q000032:analysis; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D037361:Adenylate Cyclase Toxin; D002412:Cations; D003432:Cross-Linking Reagents; D008024:Ligands; D017981:Receptors, Neurotransmitter; D017481:Receptors, Somatostatin; D010566:Virulence Factors, Bordetella; D005576:Colforsin; D004958:Estradiol; D013004:Somatostatin; D002772:Cholera Toxin; D000262:Adenylyl Cyclases