Evidence that 17α,20β,21-trihydroxy-4-pregnen-3-one is a maturation-inducing steroid in spotted seatrout

Ovarian steroidogenesis during final oocyte maturation (FOM) in the spotted seatrout (Cynoscion nebulosus) was investigated by incubating ovarian fragments with tritiated pregnenolone, followed by chromatographic separation of the radioactive products. The major tritiated steroid produced during FOM comigrated with 17α,20β,21-trihydroxy-4-pregnen-3-one (20β-dihydro-11-deoxycortisol, 20β-S) on HPLC and TLC. Only minor amounts of radioactive material coeluted with 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-P), 11-deoxycorticosterone (DOC), estradiol-17β and testosterone standards in the HPLC system. Additional chromatography by TLC confirmed the presence of radioactive estradiol-17β and testosterone but not 17α,20β-P and DOC. All the ovarian steroids producedin vitro during FOM were assayed for their ability to induce germinal vesicle breakdown (GVBD) of spotted seatrout oocytes. Twenty grams of ovarian tissue were incubated with human chorionic gonadotropin and exogenous pregnenolone. The steroidal products were purified by HPLC and TLC. Most of the maturation-inducing activity was confined to steroidal material which comigrated in these systems with 20β-S. This material was active at a concentration of 1 ng steroid/ml medium in the GVBD assay. Smaller amounts of material which coeluted with 11-deoxycortisol, DOC, 17α,20β-P and several minor unidentified fractions induced GVBD at concentrations of 10 ng steroid(s)/ml. The structure-activity relationships of authentic steroids in inducing GVBD of spotted seatrout oocytes was investigated. Hydroxylation at the 17α, 20β or 21 positions increased potency to induce GVBD. Steroids with multiple hydroxyl groups at the 17α and 20β positions (17α, 20β-P) and at the 17α, 20β, and 21 positions (20β-S) had maximum biological activity in the GVBD bioassay. The results suggest that 20β-S is a major maturation-inducing steroid in spotted seatrout.     

激素缓释制剂诱导海水养殖鱼类性腺发育成熟与生殖行为的研究进展

综述促性腺素释放激素类似物（GnRH-a)缓释制剂（SRDS），人绒毛膜促性腺激素（HCG）SRDS，类固醇激素SRDS；雄烯二酮（ADSD）和17a-甲基睾丸酮（MT），以及鲤垂体匀浆（CPE）混合SRDS诱导海水养殖鱼类性腺发育成熟和生殖行为的效果，认为采用GnRH-a SRDS处理性腺发育良好的鲈形目鱼类和鲆鲽鱼类，可以显著提高血液促性腺激素（GtH)水平，且持续时间长，诱导排卵成功，HCG乳胶SRDS可诱导日本鳗鲡（Anguila japonicus)血液GtH持续 升高，性成熟系数明显升高并且性腺发育成熟；HCG和CPE水／油／水（W／O／W）复乳SRDS对日本鳗鲡的催熟效果显著，采用ADSD和MT SRDS SRDS可成功诱导日本鳗鲡性腺成熟与排卵。     

Effects of steroid and peptide hormones on in vitro growth of previtellogenic oocytes from eel, Anguilla australis

To investigate the hormonal control of previtellogenic oocyte growth in eel, Anguilla australis, ovarian explants were incubated with or without different doses of steroid (11-ketotestosterone, estradiol-17β) or peptide hormones (ovine growth hormone, human insulin-like growth factor-I, human chorionic gonadotropin, porcine insulin) for 18 days. 11-Ketotestosterone, but not estradiol-17β, significantly increased oocyte diameters at physiological doses (30–100 ng/mL). Similarly, incubation with insulin-like growth factor-I, but not any of the other peptide hormones tested, resulted in increased oocyte diameters at high physiological doses (≥30 ng/mL). These data implicate androgens and hormones from the metabolic axis in mediating growth of previtellogenic oocytes in eel.     

Diurnal rhythm of serum steroid hormone levels in the Japanese whiting,Sillago japonica,a daily-spawning teleost

Ovarian developmental stages and serum steroid hormone levels were examined at six different times of day (0100, 0600, 1000, 1300, 1600, 2000 h) in a marine teleost, the Japanese whiting Sillago japonica, which has an asynchronous-type ovary containing oocytes at various stages of development and spawns every day during a period ranging up to three months. The largest oocytes in the ovaries at the active vitellogenic or post-vitellogenic stages were found between 0100 and 1300 h. Oocyte maturation indicated by germinal vesicle breakdown (GVBD) occurred at 1600 h, and ovulated oocytes were observed in the ovaries collected at 2000 h. These processes were accompanied by a significant daily change in serum steroid hormone levels. The serum level of estradiol-17β showed a peak in fish with mature oocytes sampled at 1600 h. In these fish, the second-largest oocytes in the ovaries were at the initial stage of vigorous vitellogenesis, the secondary yolk stage. Therefore the highest level of serum estradiol-17β was considered to be due to the second-largest oocytes. Testosterone levels remained low and constant throughout the experimental period. The serum levels of 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-diOHprog) peaked at 1600 h at which time all fish had mature oocytes. These results indicate that the Japanese whiting possesses a diurnal rhythm of oocyte development including vitellogenesis, oocyte maturation and ovulation, and further suggest that daily cycles in oocyte growth and maturation which simultaneously take place in an ovary are regulated by diurnal secretions of estradiol-17β and the maturation-inducing steroid, 17α,20β-diOHprog.     

Plasma levels of Gonadotropin-II and gonadal sex steroids in triploid catfish, Heteropneustes fossilis (Bloch)

Triploid fish have under-developed gonads due to altered reproductive endocrinology. Triploids of Indian catfish (H. fossilis) showed significantly reduced plasma levels of gonadotropin (GtH-II), testosterone (T) and estradiol-17 (E₂) than that of diploids throughout the year, except for the resting phase, irrespective of sex. Plasma levels of GtH-II were significantly different (p<0.001) between diploid and triploid fish during preparatory, prespawning and spawning phase. The plasma testosterone contents in triploids were significantly less (p<0.001) than that of diploids, except during the resting phase. Triploid females showed very low titres of estradiol-17 (<1 ng ml^–1) throughout the annual reproductive cycle in contrast to highly fluctuating levels in diploid females. Thus, this study for the first time provides information on reduced levels of GtH-II and sex steroids in plasma of male triploid fish and additional information on species-specific alteration of sex hormones in female triploids.     

Sex differences in circulating steroid hormone levels in the red drum, Sciaenops ocellatus L.

Steroid profiles of cultured and captive red drum (Sciaenops ocellatus L.) were investigated to evaluate the potential use of circulating sex steroid levels as a tool for gender identification in this species. Cultured 18‐month‐old fish were maintained on a 120‐day shortened photothermal cycle to induce precocious maturation. Additionally, wild‐caught fish were maintained in captivity under simulated natural photothermal conditions from late spring to early fall. Circulating 11‐ketotestosterone (11‐KT) levels were significantly higher in males compared with females during the early stages of gonadal growth in both cultured and captive fish. Plasma testosterone (T) levels showed a similar trend; however, the differences were significant only when males were already producing sperm. 17β‐estradiol (E₂) concentrations were low in males and females before gonadal recrudescence but increased significantly with the progression of vitellogenesis in females. These results show that a test using a minimum concentration of circulating 11‐KT could be developed to differentiate between sexes in the early stages of gonadal maturation in red drum. Moreover, plasma E₂ concentrations could be used to identify vitellogenic females. The two steroids considered together could help avoid possible error in gender identification due to unusually high levels of certain steroids encountered in some individuals.     

The influence of reproductive status on the stimulatory action of N-methyl-D,L-aspartate on growth hormone secretion, in vitro in rainbow trout, Oncorhynchus mykiss

The glutamate agonist, N-methyl-D,L-aspartate (NMA) stimulates the secretion of growth hormone (GH) from pituitary fragments in vitro and increases plasma GH levels in vivo in rainbow trout, Oncorhynchus mykiss (Flett et al. 1994; Holloway and Leatherland 1997a,b); however gonadal steroid hormones appear to modulate this response in experimental situations. This study examines whether steroid hormones also modulate the GH-regulatory actions of NMA during the normal reproductive cycle of rainbow trout by examining the relationship between the stage of sexual maturation and the pituitary release of GH in vitro in response to an NMA (10^-8 M) challenge. NMA had no effect on mean GH release from the pituitary glands of fish that were immature (GSI <1.0), from males during early development (GSI 1.0-3.0), or from sexually mature males (with free running milt) and females (ovulated). However, NMA significantly increased GH release from pituitary glands taken from females during the early stages of gonadal growth (GSI 1.0-9.0) and from males and females sampled during the later stages of gonadal growth (males GSI 3.01-6.0; females GSI 9.01-15.0). The GH-stimulatory action of NMA in males and females progressed to a maximum effect during the late stages of gonadal growth, and disappeared in ovulated females and free running males. Moreover, in female fish, the maximal GH release in response to the NMA challenge is positively correlated with plasma 17β-estradiol levels; no such correlation was evident for plasma testosterone levels in males. Changes in the GH response to NMA during maturation while gonadal steroid levels fluctuate provides further evidence to suggest that the effects of NMA on GH secretion are intimately linked to endogenous gonadal steroid hormone levels. This revised version was published online in July 2006 with corrections to the Cover Date.     

Changes in Serum Steroid Hormones and Vitellogenin Levels in Cultured Female European Eels Anguilla anguilla during Artificially Induced Ovarian Development

Changes in serum concentrations of estradiol-17β, testosterone, 17α,20βdihydroxy-4-pregnen-3-one, and vitellogenin were investigated during ovarian development induced by injection of a salmon pituitary extract in cultured European eel Anguilla anguilla . Vitellogenesis was induced with a weekly dose of 50 mg pituitary extract/kg body weight. In eels receiving that dose, gonadsomatic indices ranged from 20–43% after the 10th–11th weekly injection. Body weights were relatively stable during vitellogenesis, but increased dramatically during final maturation. Serum estradiol-17β levels increased slightly during vitellogenesis and peaked at an average of 6.95 ng/mL at final maturation. The profile of serum vitellogenin followed that of estradiol-17β which increased markedly from an average of 0.36 to 20.72 mg/mL. Control levels of vitellogenin were undetectable throughout the study. Average serum levels of testosterone increased to a peak of 17.74 ng/mL in the early stage of vitellogenesis, followed by a sharp drop to initial levels (3.86 ng/mL) in the late stage of vitellogenesis, and then increased again to an average of 8.84 ng/mL at the final maturation stage. Serum 17α,20β-dihydroxy-4-pregnen-3-one was not detected (<50 pg/mL) throughout the experiment. Profiles of serum estradiol-17β, testosterone, and vitellogenin observed during ovarian development appear different from those found in salmonids and other teleosts. This study indicates, however, that cultured European eels are a useful model for study of gonadal maturation in the eel.     

Corals have already evolved the vertebrate-type hormone system in the sexual reproduction

Most broadcast spawning scleractinian corals synchronously release gametes during a brief annual spawning period. In southern Taiwan, the mass spawning of scleractinians occurs in lunar mid-March. Endocrine system has been proposed to play important roles in this annual phenomenon. A scleractinian coral, Euphyllia ancora has been selected as a model for the hormones and reproduction studies. We detected the presence of estradiol (E2), testosterone (T), glucuronided E2, glucuronided T, aromatase, immunoreactive (ir)GnRH in coral polyps E. ancora. Annual profiles of sex steroids, aromatase, and irGnRH have also been characterized. We found the parallel increases in irGnRH concentrations, aromatase activity and free E2 and glucuronided E2 concentrations at the time of coral mass spawning. The stimulation of mammalian (m)GnRH agonist in the increased aromatase activity and sex steroid concentrations was also observed in corals. Coral extracts (irGnRH) and mGnRH agonist had a similar dose-dependent effect on luteinizing hormone release in black porgy fish pituitary cells (in vitro). In conclusion, our data suggest that irGnRH and glucuronided E2 may play important roles in the control of reproduction and mass spawning in corals. Corals already evolved the vertebrate-type hormone system in the sexual reproduction.     

The cytochrome P450 system of Atlantic salmon (Salmo salar): II. Variations in hepatic catalytic activities and isozyme patterns during an annual reproductive cycle

A group of Atlantic salmon (Salmo salar) was followed through their first year of maturation and spawning. At monthly intervals, starting with juvenile fish in December, 5–7 fish of each sex were killed, and liver and plasma were sampled. The last sampling point was of spawning fish in November a year later. Variables in the cytochrome P450 (P450) system were studied in hepatic microsomes, and estradiol 17 was measured in the plasma of females to assess the maturational status. The P450 1A1-mediated 7-ethoxyresorufin O-deethylase (EROD) started at high levels in winter, but decreased to non-detectable activities in pre-spawning females. Decreases, but not to the same extent, were also observed during this period in total cytochrome P450, cytochrome b₅, NADPH-cytochrome P450 reductase, and in the content of two immunochemically determined P450 isozymes. At the same time, LSI levels increased in maturing females (starting in July), and GSI levels increased in both sexes (starting in May). Sex specific differences were observed in pre-spawning fish in September and October, with levels of total P450, b₅, NADPH-cytochrome P450 reductase, EROD and P450 isozymes significantly lower in females. At the same time, plasma estradiol-17 levels reached peak values in females. The results point to the important role of sex steroids such as estradiol-17 as major factors in the regulation of final sexual maturation. However, this study also indicates that there may be estradiol-17 independent events of equal importance in the early stages of gonadal maturation that may involve the P450 system. The changes observed in the P450 system (as a major drug and steroid metabolizing system) of Atlantic salmon during sexual maturation may be of importance both in the endogenous transduction of hormonal signals, and as a pharmacological basis for designing therapeutic treatment of diseases in the aquaculture industry.Parts of this work were presented at the 5th International Symposium on Responses of Marine Organisms to Pollutants, April 1989 in Plymouth, United Kingdom (Larsen and Goks\/oyr 1989).     

Hormonal profile of growing male and female diploids and triploids of the blue tilapia,Oreochromis aureus,reared in intensive culture

Triploidy as a result of thermal shock exposure of fertilized eggs decreases the growth rate ofOreochromis aureus as compared to their diploid controls, but this is due to the higher female ratio present in triploids (86%) and the lower growth rate of females. When females and males are considered separately, the growth rate is not significantly different in diploids and triploids. Since triploidy results in a malfunctioning steroidogenesis in females (mainly testosterone (T) and 17β-estradiol (E₂)), but does not affect the growth rate, it is concluded that female gonadal steroids do not influence growth unless in pharmacological concentrations. These low levels of gonadal steroids are generally accompanied by higher levels of gonadotropin (GtH), but the difference is not always significant. Despite their lower growth rate diploid females have higher plasma concentrations of growth hormone (GH) during several months compared to the triploid females and diploid males. 3,5,3′-triiodo-L-thyronine (T₃) levels, however, are comparable between diploid and triploid females (except for 1 month), but higher in diploid males in 4 of the 5 months studied. 11-ketotestosterone (11kT) is always higher in males. These results indicate that the higher growth rate of males may be related to the high circulating levels of T₃ and 11kT.     

A Mammalian Growth Hormone-Releasing Hormone Increases Serum Growth Hormone Levels and Somatic Growth at Suboptimal Temperatures in Tilapia

The effects of a mammalian growth hormone-releasing hormone (GHRH), PrebGRF(1–78)OH (bGHRH), on growth and serum growth hormone (GH) levels were investigated in tilapias Oreochromis mossambicus and O. niloticus × O. aureus . Fish were injected intramuscularly or implanted intramuscularly (Silastic or cholesterol implants) with distilled water, 0.1 μg/kg bGHRH, 1.0 μg/kg bGHRH, 10.0 μg/kg bGHRH, or 100.0 μg/kg bGHRH and compared to untreated controls, fish implanted with 60 mg/kg 17α-methyltestosterone (MT), or a combination of bGHRH concentrations plus either MT, 0.01 μg/kg of a thyroid hormone (T₃), or 0.01 μg/kg of a glucocorticoid (DEX). The bGHRH increased serum GH levels in tilapia maintained at suboptimal temperatures (18 C). Serum GH levels were highest (5.3–0.45 ng/mL) for fish injected with 10.0 μg/kg bGHRH. Fish implanted with a Silastic implant containing 10.0 μg/kg bGHRH had significantly higher (4.35–0.35 ng/mL) serum GH levels than those with an equivalent dosage in a cholesterol implant. The addition of MT, thyroid hormones, and glucocorticoids did not increase serum GH levels above those obtained for fish receiving bGHRH alone. Tilapia reared at suboptimal temperatures and implanted with 10.0 μg/kg and 100.0 μg/kg bGHRH had significantly greater increases in weight and length than control fish. Fish implanted with bGHRH, MT or bGHRH plus MT had significantly higher moisture and protein content, while fat and ash contents were significantly lower than controls or sham-implanted fish. Fish implanted with bGHRH or bGHRH plus MT had significantly higher gonadosomatic indices than fish implanted with MT alone, shams or non-treated controls. This study demonstrates that a mammalian GHRH stimulates release of GH, promotes somatic and gonadal growth and may affect reproductive performance in tilapia.     

Changes in the immunostaining intensities of follicle-stimulating hormone and luteinizing hormone during ovarian maturation in the female Japanese flounder

The role of gonadotropin (GTH) in the reproduction of the Japanese flounder, Paralichthys olivaceus, was studied by assessing the changes in the apparent activity of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the pituitary gland during gonadal maturation by immunohistochemical analyses. Corresponding changes in plasma levels of testosterone (T), estradiol-17β (E₂), and 17α,20β-dihydroxy-4–pregnen-3-one (DHP) were also studied. Reared fish at the early spawning to termination stages were sampled from May to August and wild fish at the previtellogenic to termination stages were caught at 3- to 4-week intervals between April and September offshore from the northern mainland of Japan by gill nets. The gonadosomatic index of the reared fish decreased from the early spawning stage to the termination stage, while that of the wild fish increased significantly from the previtellogenic stage to the early spawning stage and decreased thereafter. In the reared fish, the immunostaining intensities of FSH and LH were high during the spawning period, accompanied by high plasma levels of T, E₂, and DHP. In the wild fish, the immunostaining intensities of FSH and LH were low during the previtellogenic stage but increased during the maturing and spawning stages. These results indicate that both FSH and LH are likely associated with oocyte maturation in the Japanese flounder.     

In vivo and in vitro effects of Rhizopus extract on body growth and steroid production in masu salmon, Oncorhynchus masou Brevoort

This study investigated the in vivo and in vitro effects of Rhizopus (filamentous fungus) extract (RU) in masu salmon Oncorhynchus masou Brevoort. Underyearling fish were fed with RU for 16 months. Monthly changes in body growth, gonadal maturation and serum levels of sex steroids were monitored. Gonads were also incubated at 0, 1, 10, 100 and 1000 μg RU mL⁻¹ Leibovitz's L‐15 medium for 18 h. The levels of steroids in serum and cultured medium were measured. It was determined that RU‐fed immature and mature males, when compared with control groups, showed significantly higher body growth during spring, summer and the spawning period. Similarly, immature RU‐fed females showed significantly higher fork length and body weight in autumn, spring and summer. Furthermore, RU‐fed males showed significantly higher levels of serum testosterone (T) and 11‐ketotestosterone (11‐KT) levels in the pre‐spawning season, and 17α, 20β‐dihydroxy‐4‐pregnen‐3‐one (DHP) in the spawning season. In vitro RU incubation of gonads showed a dose‐dependent and significant increase in T, 11‐KT, oestradiol‐17β and DHP release in the medium. It appears that the causes of enhanced body growth and increased steroid production herein observed in salmonids are the physiologically active substances that may be present in the mycelium of the fungus.     

Changes in gonadal hormones during oocyte development in the striped bass,Morone saxatilis

Wild striped bass,Morone saxatilis, were collected from coastal waters and spawning areas to describe the endocrine correlates of oocyte development in non-captive, migratory fish. The fish were classified according to their most advanced oocytes. Serum levels of estradiol (E₂), testosterone (T) and 17-20-dihydroxyprogesterone (DHP) were measured by radioimmunoassay (RIA). Females in the primary growth phase and early secondary growth phase (pre-vitellogenic) had low levels of plasma steroids, ovarian lipid content and gonadosomatic indices (GSIs). Significant increases in E₂, T, ovarian lipid content and GSIs occurred during the vitellogenic phase. Maximum levels of all reproductive parameters were found in prespawning fish sampled in the Hudson River. Mean levels of E₂, T, ovarian lipids and GSIs for these fish were 2.0±0.5 ng/ml, 3.0±0.3 ng/ml, 24±1 mg/g, and 5.6±0.3% (mean±SEM), respectively. In fish induced to spawn with human chorionic gonadotropin (HCG), DHP levels (1.9±0.4 ng/ml) were significantly elevated. Similar levels were found in two fish captured during the spawning season, suggesting that DHP may serve as the maturation-inducing steroid in this species.     

Effect of temperature decrease on oocyte development,sex steroids,and gonadotropin β-subunit mRNA expression levels in female Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

To improve understanding of the mechanism of early ovarian development in eels, the effects of water temperature decrease on oocyte development, plasma levels of sex steroids [estradiol 17β (E2), testosterone (T), 11-ketotestosterone (11-KT)], and gonadotropin β-subunit [follicle-stimulating hormone (FSHβ), luteinizing hormone (LHβ)] messenger RNA (mRNA) expression levels were investigated. A total of 27 female Japanese eels Anguilla japonica were divided into initial, control, and test (water temperature decrease) groups. Starting on 22 September 2009, eels in the test group were reared in a tank with gradual temperature decrease from 25°C to 15°C over 39 days, while the control group was maintained at 25°C. The test group accumulated more oil droplets in their oocytes than did the other groups. Levels of sex steroids, especially 11-KT, were higher in the test group. In contrast, FSHβ and LHβ mRNA expression levels were lower in the test group. These results suggest that water temperature decrease only induced an early stage of ovarian development that was partly affected by an 11-KT increase. For further maturation, other environmental factors related to induction of gonadotropin increase appear to be needed.     

Effects of steroid hormones on immunoglobulin M (IgM) in rainbow trout, Oncorhynchus mykiss

The immunosuppressive effects of steroid hormones were evaluated as the response against implanted steroid hormones, cortisol (F), testosterone (T), estradiol-17 (E2), and 11- ketotestosterone (11-KT), in juvenile rainbow trout. In long term experiments (5 weeks), fish were given a single intraperitoneal implant of F or T. A clear suppressive effect of plasma IgM levels with F and T was not necessarily obtained, although mucus IgM levels were reduced corresponding to the elevated plasma steroid hormone levels. In short term experiments (1 week), intraperitoneal implantation of T, 11-KT and E2 suppressed plasma and mucus IgM levels, although the effects were not dose-dependent. When administered through diet, F and T caused a suppression of plasma IgM levels; F administration at both high and low dosages caused a significant decrease in plasma IgM levels, while only a high dose of T caused the suppression. These results suggest that sex steroid hormones, as well as F, have immunosuppressive functions in rainbow trout.     

The maturation of the salmon GnRH system and its regulation by gonadal steroids in masu salmon

Pituitary gonadotropin (GTH) secreting cells and brain gonadotropin-releasing hormone (GnRH) secreting neurons are known to be subjected to feedback control by gonadal steroid in teleosts. In masu salmon, Oncorhynchus masou, salmon GnRH (sGnRH) neurons in the ventral telencephalon (VT) and the preoptic area (POA) are involved in the control of GTH cells because sGnRH synthesis in these areas is activated with gonadal maturation. In this study, we attempted to clarify mechanisms of feedback control of sGnRH neurons by gonadal steroids. We examined the effects of 17-methyltestosterone (MT) on sGnRH synthesis in yearling and 2-year-old female fish (which were immature during experimentation in May), and the effects of castration on sGnRH synthesis in underyearling precocious male fish in August. sGnRH synthesis in the POA, but not in the VT, was increased by MT administration in 2-year-old females only, indicating higher sensitivity to MT in the preoptic sGnRH neurons. Castration increased sGnRH synthesis in the VT but not in the POA. These results suggest that sGnRH neurons in the VT and those in the POA are differentially regulated by gonadal steroids.     

Changes in Plasma Concentrations of Sex Steroids in Adult Pacific Halibut, Hippoglossus stenolepis

Blood samples were collected from captive Pacific halibut, Hippoglossus stenolepis , at intervals of about six weeks from early December 1986 to late November 1987. Concentrations of plasma androgen and estradiol-17β were determined by radioimmunoassay. The plasma concentrations of steroid were highest during autumn and winter in halibut that matured during late winter. The concentrations of steroids in samples collected in December were above 2 ng/mL (estradiol) or 1 ng/mL (androgen) in maturing females and below 0.5 ng/mL for both steroids in non-maturing females. The levels of steroids decreased rapidly about one month before spawning. In a mature male, androgen began to rise in August and November, and reached a peak of 7 ng/mL in early December. One month before spawning, the androgen concentration fell to 0.16 ng/mL. Estradiol concentrations were detectable in the male and varied little during the year. In immature fish, neither androgen nor estradiol changed significantly throughout the year. These results suggest that the concentrations of estradiol or androgen measured in blood samples taken during December may be used to determine the sex and state of maturation of Pacific halibut.     

Hormonal and pheromonal control of spawning behavior in the goldfish

Species that employ sexual reproduction must synchronize gamete maturity with behavior within and between genders. Teleost fishes solve this challenge by using reproductive hormones both as endogenous signals to synchronize sexual behavior with gamete maturation, and as exogenous signals (pheromones) to synchronize spawning interactions between fish. This dual role of hormonal products is best understood in the goldfish, an external fertilizer with a promiscuous mating system. Female gonadal growth and vitellogenesis is stimulated by 17β-estradiol (E2) which also evokes release of a recrudescent pheromone. At the completion of vitellogenesis, ovarian E2 production drops and plasma testosterone increases, sensitizing the female gonadotropin II (luteinizing hormone; LH) system to environmental cues (temperature, spawning substrate, pheromones). These cues eventually trigger a LH surge that alters steroidogenesic pathways to favor the production of progestins including 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P). Plasma 17,20β-P stimulates oocyte maturation but is also released to the water along with sulfated 17,20β-P and androstenedione to serve as a preovulatory pheromone. This pheromone stimulates male behavior, LH release, and sperm production. At the time of ovulation, females become sexually active in response to prostaglandin F2α (PGF2α) synthesized in the oviduct. PGF2α and its metabolites are released as a postovulatory pheromone that induces male spawning behavior which further increases male LH and sperm production. Androgenic hormones are required for male behavior and LH release. Although goldfish are gonochorists, hormone treatments can induce heterotypical functions in adults. Similar findings in other fish demonstrate that a sexually bipotential brain is not restricted to hermaphroditic fishes. This revised version was published online in July 2006 with corrections to the Cover Date.