title: Receptors and secretory actions of sigma/phencyclidine agonists in anterior pituitary cells.
abstract: Opiate receptor subtypes in the adenohypophysis were analyzed by binding studies with tritiated etorphine, phencyclidine (PCP), and N-allylnormetazocine [(+)SKF 10,047] in anterior pituitary cell (AC) cultures and membranes, and in cell populations separated by centrifugal elutriation. In cultured AC, specific binding of [3H]etorphine revealed two sets of saturable sites with Kd values of 5 nM and about 10 microM. The high affinity [3H]etorphine sites were present in low concentration and represent specific opiate receptors that mediate the direct inhibitory actions of etorphine and morphine on LH release in vitro. The more abundant low affinity sites, observed in the presence of higher concentrations of unlabeled opiates, exhibited the properties of sigma/PCP receptors. In intact AC and pituitary membranes, specific [3H]PCP binding was saturable with respect to labeled and unlabeled ligand concentrations, and Scatchard analysis revealed a single class of relatively high affinity [3H]PCP-binding sites (Kd = 98 nM in pituitary membranes). Relative potencies derived from inhibition of [3H]PCP binding in AC by PCP-related drugs were: (-) cyclazocine greater than dexoxadrol greater than N-[1-(2-Thienyl)cyclohexil]piperidine greater than PCP greater than (+)SKF 10,047 greater than levaxodral greater than (+)cyclazocine less than (-)SKF 10,047 greater than (+)ethylketocyclazocine greater than haloperidol greater than (-)ethylketocyclazocine. In elutriated pituitary cells, specific [3H]PCP binding was correlated with the LH content of the individual cell fractions. The binding of (+)-[3H]SKF 10,047 was also specific and saturable in AC and anterior pituitary membranes, which contained two classes of binding sites with Kd values of 87 nM and 3.3 microM. In fractionated pituitary cells, specific binding of (+)-[3H]SKF 10,047 was similar in enriched lactotrophs and gonadotrophs. The high affinity class of (+)-[3H]SKF 10,047-binding sites probably corresponds to sigma-receptors, and the low affinity class to PCP receptors. In contrast to the inhibitory actions of opiates on LH release in vitro, PCP and (+)SKF 10,047 stimulated LH release in cultured AC and enhanced the secretory responses to GnRH as well as KCl. The stimulation of LH release by PCP was dependent on extracellular calcium and is probably related to increased transmembrane calcium influx. The stimulatory sites may correspond to selective sigma/PCP receptors, and could represent a distinct nonopiate receptor subtype with the potential for modulation of gonadotropin secretion.
authors.affiliation: Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.
authors.forename: S S
authors.initials: SS
authors.lastname: Stojilković
authors.forename: M L
authors.initials: ML
authors.lastname: Dufau
authors.forename: K J
authors.initials: KJ
authors.lastname: Catt
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D005048:Etorphine / Q000378:metabolism; D005260:Female; D007700:Kinetics; D007986:Luteinizing Hormone / Q000378:metabolism; D010620:Phenazocine / Q000031:analogs & derivatives* / Q000378:metabolism; D010622:Phencyclidine / Q000378:metabolism* / Q000494:pharmacology; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D017981:Receptors, Neurotransmitter / Q000187:drug effects / Q000502:physiology; D011957:Receptors, Opioid / Q000187:drug effects / Q000502:physiology*; D017478:Receptors, Phencyclidine; D017480:Receptors, sigma; D013816:Thermodynamics
chemical_list: D017981:Receptors, Neurotransmitter; D011957:Receptors, Opioid; D017478:Receptors, Phencyclidine; D017480:Receptors, sigma; D005048:Etorphine; C012134:SK&F 10047; D007986:Luteinizing Hormone; D010620:Phenazocine; D010622:Phencyclidine; D011188:Potassium