ay-25-205 and 11-ketotestosterone

Spermiating male plaice were caught in the North Sea and acclimatised to laboratory conditions. In two experiments, males were injected intramuscularly with either microspheres or pellets containing gonadotrophin-releasing hormone agonist (GnRHa). Blood was sampled at 2- to 5-day intervals. Individual blood plasma specimens were assayed for testosterone, 5beta-reduced, 3alpha-hydroxy ("5beta,3alpha") steroids and sulphated 17, 20beta-dihydroxy ("17,20beta") steroids. Pooled plasma samples were also assayed for free and sulphated 17, 20beta-dihydroxy-4-pregnen-3-one, free 11-ketotestosterone, and glucuronidated testosterone and 11-ketotestosterone. Plasma concentrations of all steroids were significantly elevated by GnRHa from 2 to 5 days onwards following treatment. The most marked changes occurred in the concentrations of the sulphated 17,20beta steroids, which comprised approximately equal amounts of 5beta-pregnane-3alpha,17,20beta-triol 20-sulphate (3alpha,17, 20beta-P-5beta-S) and 5beta-pregnane-3beta,17,20beta-triol 20-sulphate, rising from ca. 1 to 30-80 ng/ml in the first and from ca. 8 to 80 ng/ml in the second experiment. Concentrations of 5beta, 3alpha steroids matched those of 17,20beta steroids in one experiment. However, in the other experiment, the two RIAs yielded highly disparate results in about 50% of the fish (including males in the control group). The plasma of these fish contained excessive amounts of 5beta,3alpha-immunoreactive material between 10 and 25 days. This material was identified as 3alpha,17, 21-trihydroxy-5beta-pregnan-20-one 21-sulphate (a metabolite of 11-deoxycortisol). All previous studies have indicated that when plasma concentrations of this steroid are high, so are those of 3alpha,17,20beta-P-5beta-S. This is the first indication that these steroids are regulated independently. In a third experiment, milt fluidity and production were assessed at 10, 15, and 25 days following GnRHa implantation. Milt volume and fluidity were significantly enhanced by the GnRHa treatment.

Topics: Animals; Chromatography, High Pressure Liquid; Cortodoxone; Delayed-Action Preparations; Dose-Response Relationship, Drug; Female; Flounder; Glucuronates; Gonadotropin-Releasing Hormone; Hormones; Hydroxylation; Hydroxyprogesterones; Male; Microspheres; Oxidation-Reduction; Radioimmunoassay; Semen; Steroids; Sulfates; Testosterone; Viscosity

The effects of gonadal steroids on gonadosomatic index (GSI; gonad wt/total body wt x 100), pituitary gonadotropin (GTH) content, and serum GTH response to [D-Ala6,Pro9-Net]-luteinizing hormone-releasing hormone (LHRH-A) were investigated throughout the seasonal reproductive cycle of the goldfish. Gonad-intact female fish were implanted i.p. for 5 days with silastic pellets containing no steroid (blank), testosterone (T; 100 micrograms/g), or estradiol (E2; 100 micrograms/g). The serum GTH response at 6 h following i.p. injection of saline or 0.1 microgram/g LHRH-A was assessed. In blank-implanted, saline-injected animals, seasonal variations in GSI, pituitary GTH content, and serum GTH levels were evident; maximal and minimal levels were noted in the spring and summer months, respectively. In blank-implanted fish, LHRH-A effectively stimulated GTH release in females undergoing gonadal recrudescence (late autumn and winter) and in sexually mature (spring) females, but not in sexually regressed (summer and early autumn) females. Implantation of T or E2 raised serum steroid levels to those found during ovulation in goldfish. Steroid treatments did not affect unstimulated serum GTH levels at any time of the year. Testosterone effectively potentiated the serum GTH response to LHRH-A during the entire reproductive cycle, whereas the positive effects of E2 were evident in sexually regressed and post-spawning females only. Both T and E2 potentiated the GTH response to LHRH-A in male fish. To examine the involvement of T aromatization in mediating its actions on induced GTH secretion, male and female fish were implanted with T or the nonaromatizable androgens 5 alpha-dihydroxytestosterone (DHT; 100 micrograms/g) and 11-keto-testosterone (11-KT; 250 micrograms/animal). Testosterone potentiated the GTH response to LHRH-A in both males and females whereas DHT and 11-KT were without effect. Furthermore, the positive action of T on induced GTH secretion was blocked by 2-day pretreatment with the aromatase inhibitor 1,4,6-androstatrien-3,17-dione (100 or 300 micrograms/g). Multiple i.p. injections of hCG (0.2 microgram/g every 3 days for 39 days), probably through stimulation of endogenous T secretion, resulted in potentiation of the GTH response to LHRH-A in mature male goldfish. These results clearly demonstrate that T, through aromatization to E2, can increase pituitary responsiveness to exogenous LHRH-A in gonad-intact male and female goldfish.

Topics: Animals; Dihydrotestosterone; Drug Synergism; Estradiol; Female; Goldfish; Gonadotropin-Releasing Hormone; Gonadotropins, Pituitary; Hormones; Male; Reproduction; Seasons; Testosterone