Effects of temperature on the final stages of sexual maturation in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.)

Maturing male and female Atlantic salmon (Salmo salar L.) were held under three temperature regimes for 10 weeks between September and December: warm (constant 14–16 °C), ambient (decreasing from 11 to 5 °C), and cold (decreasing from 7 to 3 °C). Blood samples were analyzed for plasma steroid levels, and the fish were inspected for the presence of expressible milt (total volume and spermatocrit) and ovulation weekly. Samples of eggs were dry-fertilized with milt stripped from three males held at the same temperatures and incubated until the eyed stage. In females, levels of plasma testosterone (T) and 17β-oestradiol (E₂) dropped as ovulation approached, concurrent with a rapid increase in levels of plasma 17α,20β-dihydroxy-4-pregnen-3-one (17,20β-P). In males, levels of T and 11-ketotestosterone (11-KT) peaked 2–3 weeks after the first appearance of expressible milt, while levels of 17,20β-P increased steadily and did not exhibit a definite peak. Exposure of females to cold water amplified and advanced the profiles of all three steroids compared with the ambient group, and increased the survival rates to the eyed egg stage. Cold water had no immediate effect on the male steroid profiles, but later, higher levels of 17,20β-P were evident compared with both the ambient controls and the warm water group, while the effects on 11-KT and T were more variable. Exposure to warm water completely inhibited both milt production and ovulation. Moreover, warm water modulated the steroid profiles of the males with lower 11-KT levels compared with ambient controls and lower 17,20β-P level compared with cold-water-treated males. In females, warm water resulted in total inhibition of the peri-ovulatory peak in 17,20β-P and prevented the normal decline of T and E₂ levels associated with ovulation. The findings of the present study are highly relevant for broodstock management in aquaculture, as well in understanding the impact of climate change/temperature variability on wild salmon spawning.     

Plasma levels of immune factors and sex steroids in the male seahorse <Emphasis Type="Italic">Hippocampus erectus</Emphasis> during a breeding cycle

To better understand the endocrine- and immune-response pattern during reproduction in a fish species having parental care behaviors and also to accumulate the endocrine- and immune-related data for future explanations of the low reproductive efficiency in seahorse species, the variations of immune factors and sex steroids in the plasma of the male lined seahorse Hippocampus erectus at different breeding stages, i.e., pre-pregnancy, pregnancy (early, middle, and late periods), and post-pregnancy, were investigated in the present study. The immune factors included monocytes/leucocytes (M/L), leucocyte phagocytic rate (LPR), immunoglobulin M (Ig M), interleukin-2 (IL-2), interferon-α (IFN-α), and lysozyme (LZM). The sex steroids included testosterone (T), 11-ketotestosterone (11-KT), 11β-hydroxytestosterone (11β-OHT), 17α-methyltestosterone (17α-MT), 17β-estradiol (E2), and 17α-hydroxy-20β-dihydroprogesterone (17α-20β-P). Moreover, the immune metabolic activity of epithelium cells in the brood pouch at different breeding stages was also analyzed through ultrastructural observations of the abundance of cytoplasmic granules, mitochondria, endoplasmic reticulum, lysosomes, and exocytosis. The results show that a higher immune level was observed during pregnancy, particularly in the early and middle periods, and a lower immune level was noted during pre-pregnancy. Correspondingly, the epithelium cells in the brood pouch also showed a stronger immune metabolic activity during pregnancy and weaker activity during pre-pregnancy. Four sex steroids of T, 11β-OHT, 17α-MT, and E2 were higher during pre-pregnancy and lower during post-pregnancy, whereas 11-KT and 17α-20β-P, which were positively correlated with part immune factors, were higher during pregnancy. No negative correlations between sex steroids and immune factors were observed. In conclusion, the higher immune competence during pregnancy may indicate that parental care could improve immunity, which may be the major factor for no immunosuppressive effect of sex steroids during reproduction in the seahorse H. erectus, unlike noncaregiving fishes in which inhibitions of sex steroids on immunity are frequently observed. Moreover, higher 11-KT and 17α-20β-P during pregnancy than during pre-pregnancy and post-pregnancy may suggest that these two steroids are also involved in parental care regulation.     

Hormonal changes accompanying sexual maturation in captive milkfish (Chanos chanos Forsskal)

Steroid hormone profiles accompanying sexual maturation in captive milkfish are described. There were no significant differences in levels of serum estradiol 17- (E₂) and testosterone (T) between immature male and female fish. Mean E₂ levels rose from 0.54±0.11 ng/ml in immature females (Stage 1) to 4.53±1.16 ng/ml in vitellogenic females (Stage 5), while T levels increased from 2.06±0.28 ng/ml to 38.4±9.26 ng/ml. E₂ and T levels were positively correlated to GSI and oocyte diameter. In males, serum T levels increased from 2.5±0.40 ng/ml in immature males to 27.73±5.02 ng/ml in spermiating males. A significantly higher T level was found in males with thick and scantly milt (spermiation index, SPI, 2) compared to males with scanty milt (SPI, 1) or males with copious, fluid milt (SPI, 3).Serum levels of E₂ and T, and the GSI in females rose significantly during the breeding season (April–June 1983). The levels of both steroids dropped below 1 ng/ml in spent females sampled in succeeding months. In immature males, T levels ranged from 1.11 ng/ml to 2.78 ng/ml and rose significantly to 21.52±8.38 ng/ml during the breeding season when GSI peaked. Serum T levels dropped to around 10 ng/ml in the succeeding months when only spent or regressed males were sampled.     

Molecular control mechanisms of fish spermatogenesis

Fish spermatogenesis, from spermatogonial stem-cell renewal to sperm maturation, is controlled by the sex steroid hormones. Mitotic divisions of spermatogonia can be categorized by spermatogonial stem cell renewal and spermatogonial proliferation. Spermatogonial renewal is regulated by estradiol-17β (E₂; the natural estrogen in vertebrates), and spermatogonial proliferation toward meiosis is promoted by 11-ketotestosterone (11-KT), the main androgen in teleost. The action of E2 and 11-KT is mediated by other factors produced by Sertoli cells; E2 is mediated by spermatogonial stem-cell renewal factor and 11-KT is mediated by spermatogenesis preventing substance and activin B. Although 11-KT also induce meiosis and spermiogenesis, the control mechanisms of these processe are not clear. After spermiogenesis, immature spermatozoa undergo sperm maturation. Sperm maturation is regulated by 17α,20β-dihydroxy-4-pregnen-3-one (DHP), which is progestin in teleost. The DHP acts directly on spermatozoa to active the carbonic anhydrase existed in the spermatozoa. This enzymatic activation causes an increase in the seminal plasma pH, enabling spermatozoa to motile.     

Endocrine and gonadal changes during the annual reproductive cycle of the freshwater teleost,Stizostedion vitreum

The annual reproductive cycle of walleye (Stizostedion vitreum) was characterized by documenting changes in gonadal development and serum levels of estradiol-17β (E₂), testosterone (T), 17α,20β-dihydroxy-4-pregnen-3-one (17,20-P), and 11-ketotestosterone (11-KT) in wild fish captured from upper midwestern lakes and rivers throughout the year. Fish from the populations used in this study spawn annually in early- to mid-April. Walleye showed group synchronous ovarian development with exogenous vitellogenesis beginning in autumn. Oocyte diameters increased rapidly from ∼ 200 μm in October to ∼ 1,000 μm in November, and reached a maximum of 1,500 μm just prior to spawning. Changes in gonadosomatic indices (GSIs) paralleled changes in oocyte diameters. Serum E₂ levels in females increased rapidly from low values in October (< 0.1 ng ml⁻¹) to peak levels of 3.7 ng ml⁻¹ in November, coinciding with the period of the most rapid ovarian growth. Subsequently, E₂ levels decreased from December through spawning. Serum T levels exhibited a bimodal pattern, increasing to 1.6 ng ml⁻¹ in November, and peaking again at 3.3 ng ml⁻¹ just prior to spawning. We detected 11-KT in the serum of some females at concentrations up to 5.6 ng ml⁻¹, but no seasonal pattern was apparent. In this study (unlike our results in a related study) 17,20-P was not detected. In males, differentiation of spermatogonia began in late August, and by January the testes were filled (> 95% of germ cells) with spermatozoa. Mature spermatozoa could be expressed from males from January through April. GSIs ranged from 0.2% (post-spawn) to 3.2% (pre-spawn). Serum T levels rose from undetectable levels in post-spawn males to 1.6 ng ml⁻¹ by November, remained elevated throughout the winter, and peaked at 2.8 ng ml⁻¹ I prior to spawning. Levels of 11-KT in males remained low (< 10 ng ml⁻¹, from post-spawning through January, then increased significantly by March and peaked just prior to spawning at 39.7 ng ml⁻¹. Our results indicate that vitellogenesis and spermatogenesis are complete or nearly so, in walleye by early winter, and suggest that it may be possible to induce spawning in this species several months prior to the normal spawning season by subjecting fish to relatively simple environmental and hormonal treatments.     

Estradiol-17β suppresses testicular development and stimulates sex reversal in protandrous black porgy,Acanthopagrus schlegeli

Two year old black porgy (Acanthopagrus schlegeli) fed a diet containing 4.0 mg kg^–1 of estradiol-17 (E₂) for 5 months had significantly lower GSI than the control group during the spawning season. E₂ suppressed testicular development, spermiation and plasma testosterone (T) and 11-ketotestosterone (11-KT) and stimulated ovarian development, vitellogenesis and sex reversal. Spermiation in the control group occurred in January and February with the concentrations of 1.08–1.36 × 10¹⁰ sperm ml^–1 of milt. Higher plasma T and 11-KT, but lower E₂ levels were detected in the spermiating fish (control group). Higher plasma E₂ levels were detected in the sex reversing black porgy during the pre-spawning season. A sharp rise in plasma 11-KT and a drop in T levels were detected in spermiating fish (control group) from January to February. Plasma 11-KT levels correlated with the testicular development and spermiation. The data suggest that E₂ plays an important role in controlling the sex reversal of black porgy.to whom correspondence should be addressed.     

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.     

Involvement of sex steroids,luteinizing hormone and thyroid hormones in upstream and downstream swimming behavior of land-locked sockeye salmon <Emphasis Type="Italic">Oncorhynchus nerka</Emphasis>

The involvement of testosterone (T), estradiol-17β (E₂), 11-ketotestosterone (11-KT), 17,20β-dihydroxy-4-pregnene-3-one (DHP), luteinizing hormone (LH), thyroxine (T₄), and triiodothyronine (T₃) in the regulation of downstream and upstream movement (swimming behavior) was investigated in land-locked sockeye salmon Oncorhynchus nerka, using an artificial raceway. During the downstream migratory period, T implant resulted in high plasma T levels and inhibited the occurrence of downstream swimming behavior (negative rheotaxis) in yearling (1+) immature smolts. In terms of upstream behavior, 2-year-old (2+) males exhibited high plasma T and 11-KT levels, while 2+ females had elevated T and DHP levels. In 1+ immature fish, a T implant induced upstream swimming behavior (positive rheotaxis). In experiments 1 and 3, the plasma T₄ and T₃ levels of non-migrants tended to be higher than those of migrants. In contrast, no marked changes in plasma and pituitary LH were found in both downstream and upstream migrants. These results suggest that sex steroids, such as T, play significant roles in the regulation of downstream and upstream swimming behaviors in land-locked sockeye salmon.     

Seasonality of reproduction in male spotted murrel <Emphasis Type="Italic">Channa punctatus</Emphasis>: correlation of environmental variables and plasma sex steroids with histological changes in testis

The present study was undertaken to develop a comprehensive understanding of how environmental cues and sex steroids relate with cyclic changes in spermatogenesis in freshwater spotted snakehead Channa punctatus that is nutritious and economically important. The seasonal histological changes in testis and annual profile of gonadosomatic index (GSI) of C. punctatus delineated the testicular cycle into four phases: regressed (December–March), preparatory (April–June), spawning (July and August) and postspawning (September–November). Among environmental variables, correlation and regression analyses exhibited an important relationship between photoperiod and testicular weight while role of rainfall was seen confined to spawning. The seasonal profile of plasma sex steroids when correlated with cyclic changes in spermatogenesis in spotted snakehead, testosterone (T) seems to be involved in controlling the major events of spermatogenesis from renewal of stem cells to spawning of spermatozoa. Another important androgen prevalent in teleosts, 11-ketotestosterone (11-KT), was high during preparatory phase, suggesting that 11-KT in addition to T plays an important role in progression of spermatogenesis and spermiation in C. punctatus. However, 11-KT was not seen to be associated with milt production and release of spermatozoa during spawning. Plasma profile of estradiol-17β (E₂) during different reproductive phases revealed the involvement of E₂ in repopulation of stem cells during postspawning phase and in maintaining quiescence of testis during regressed phase.     

Testicular development and serum profiles of steroid hormone levels in captive male Pacific herring Clupea pallasii during their first maturational cycle

The present study investigates the relationship between testicular development and serum steroid hormone levels in captive Pacific herring Clupea pallasii during the first reproductive cycle. The maturity of the testis was divided into five periods based on histological observation. These are early spermatogenic stage (April to July), mid-spermatogenic stage (August to November), late spermatogenic stage (December to March), functional maturation stage (early April) and spent stage (late April). The pattern of seasonal change in gonadosomatic index (GSI) clearly reflected testicular maturity. 11-Ketotestosterone (11-KT) levels increased from October to a peak level (6.58 ± 1.87 ng/mL) in January, and were maintained at this level until March. In contrast, testosterone levels were consistently low, less than 1 ng/mL, at all times. These results suggest that 11-KT is the predominant androgen that controls spermatogenesis in this species. 17,20β-Dihydroxy-4-pregnen-3-one (DHP) showed a single sharp peak (3.38 ± 0.35 ng/mL) in early April of the second year, suggesting that milt production is induced by DHP as in some other teleost species.     

鲤鱼血清中促性腺激素、17β雌二醇含量的周年变化

应用放射免疫测定法,对雌、雄鲤血清中促性腺激素(GTH)和17β-雌二醇(17β-E_2)的含量进行周年测定。GTH的常年分泌量,雌鲤平均为8.94±5.94ng/ml(12月份含量最低,峰值在1月份);雄鲤常年平均为8.72±5.20ng/ml(10月份含量最低,峰值在11月份)。17β-E_2的含量,雌鲤1-4月份较高,平均为2019.48±754.49pg/ml(全年峰值在2月份)。产卵季节后,卵巢开始退化,17β-E_2含量下降。5—8月平均为454.58pg/ml,产卵前后差异性显著。这表明性腺的发育和成熟与血清中17β-E_2含量变化的密切相关性以及雌二醇对GTH分泌的反馈抑制作用。     

Reproductive performance of common dentex, Dentex dentex, broodstock held under different photoperiod and constant temperature conditions

The effect of photoperiod and temperature on the timing and the quality of spawning, and on associated endocrine changes in circulating 17β-oestradiol, 11-ketotestoserone (11-KT) and vitellogenin (Vtg) were investigated in common dentex, Dentex dentex, undergoing their second reproductive cycle. The possibility was also explored of using the measurement of steroids in the culture water of broodstock tanks, rather than in individual blood plasma samples, as a potentially useful tool for assessing the physiological state of a fish without disturbing them. One group of fish was exposed to a simulated natural seasonal cycle and ambient temperature (CONTROL). The other two groups were exposed to simulated seasonal photoperiod cycles (12 month-long) but which were phase-shifted either three months before (ADVANCED) or after (DELAYED) the natural cycle. Temperature was kept at 19.4±0.9 °C all-year-round. In the CONTROL, spawning started in mid-April and lasted until mid-June, while in the ADVANCED group, spawning started 4 months earlier and in the DELAYED group 2 months later than the CONTROL. The total egg production, egg quality, hatching rate, relative fecundity, and spawning index of the experimental groups were similar to the controls. The differences in spawning time induced by photothermal manipulation were associated with a difference in the timing of peak concentrations of plasma E₂, 11-KT and Vtg. In all three groups, the amounts of conjugated 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P) and free and sulfated 11-KT which could be extracted from the water during the spawning period were significantly higher than those found in the preovulatory period. However, the differences were mostly less than 2-fold suggesting that, at least under the conditions employed in this study, the method was of limited use for non-intrusive detection of gonadogenesis and spawning (as had been hoped). The observed differences in spawning time and in the seasonal changes of sex steroids and Vtg confirm and extend the findings on marine fish. This revised version was published online in August 2006 with corrections to the Cover Date.     

Regulation of gonadotropin subunit genes expression by 11-ketotestosterone during early spermatogenesis in male red seabream, Pagrus major

To examine the roles of gonadal steroids in the regulation of expression of gonadotropin subunit genes, male red seabream were gonadectomized and a sub-group was treated with 11-ketotestosterone (11-KT). Castration of males during the early stage of spermatogenesis elicited a significant increase in FSHβ mRNA levels, which was prevented by 11-KT replacement. By contrast, LHβ mRNA levels were not changed by castration or 11-KT replacement. In addition, administration of 11-KT to sham-operated males suppressed the steady-state FSHβ and LHβ mRNA levels. These results indicate that 11-KT may function as a negative feedback regulator of FSHβ gene expression, and may act through the testis to down-regulate LHβ mRNA levels in male red seabream during this period.     

Gonad histology and plasma steroid profiles in wild New Zealand freshwater eels (Anguilla dieffenbachii and A. australis) before and at the onset of the natural spawning migration. I. Females*

To determine steroid profiles in immature and maturing female eels from the wild, non-migratory and migratory New Zealand longfinned (Anguilla dieffenbachii) and shortfinned (A. australis) eels were caught and blood and ovarian samples collected. Plasma steroid levels were determined and related to the developmental stage of the ovary. Ovaries of non-migrants contained oogonia and previtellogenic oocytes. Vitellogenic oocytes were never observed in these groups, but instead were very common among migrants (up to 88% of oocytes). Concentrations of both androgens (androstenedione (AD), testosterone (T)) and estradiol-17 (E2) were higher in migrants than in non-migrants. Among migrants, T levels were higher in shortfins (2.27 ± 0.14 ng ml–1) than in longfins (0.82 ± 0.10 ng ml–1), whereas E2 levels were higher in longfins (mean 2.46 ng ml–1) than in shortfins. Levels of sex steroids were generally low in non-migrants. In contrast, plasma levels of 17-hydroxyprogesterone were significantly higher in non-migrants than in migrants. Similarly, cortisol levels were higher in non-migrating than in migrating shortfinned, but not longfinned, females. 17,20-Dihydroxy-4-pregnen-3-one, the putative maturation-inducing steroid in anguillids, was near minimum-detectable levels for all animals examined. Surprisingly, very high levels of 11-ketotestosterone (KT) were found in migrants, averaging nearly 3 ng ml–1 in longfins and over 20 ng ml–1 in shortfins. The identity of KT and several 5-reduced androgens was confirmed using gas chromatography - mass spectrometry. The function of KT in females is not known, but we suggest that this steroid hormone may play a role in preparing maturing animals for their spawning migration.     

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.     

In vitro metabolism of progesterone, androgens and estrogens by rainbow trout embryos

The purpose of the study was to determine the steroid metabolic pathways used by rainbow trout (Oncorhynchus mykiss) embryos. Whole embryo preparations at 42, 53 and 65 days post fertilization (dpf) were incubated, in vitro, with the tritium-labelled steroids, progesterone (P₄), testosterone (T), androstenedione (A4), 17β-estradiol (E₂), and estrone (E₁), and the metabolites formed were separated and identified by reverse phase high performance liquid chromatography (HPLC). The degree of metabolism for each of the substrates was dependent on the age of the embryos, and was always higher in older embryos. P₄ yielded three metabolites, one of them identified by gas chromatography-mass spectroscopy as 3β,7α-dihydroxy-5α-pregnan-20-one. Several unknown metabolites of a similar profile on HPLC were produced for T and A₄, in addition to A₄ and T as metabolites of T and A ₄ metabolism, respectively. Likewise, E₂ and E₁ yielded each other and an unknown polar metabolite. These observations argue for the presence of 5α-reductase, 7α-hydroxylase and 17β-hydroxysteroid dehydrogenase activity in embryo tissues which, by their conversion of biologically active yolk steroids of maternal origin, may provide a protective environment for the embryo during early development.     

Diel cyclic hypoxia alters plasma lipid dynamics and impairs reproduction in goldfish (<Emphasis Type="Italic">Carassius auratus</Emphasis>)

Diel cyclic hypoxia occurs with varying frequency and duration in freshwater habitats, yet little is known about its effects on reproduction of freshwater fishes. The present study shows that long-term exposure of goldfish (Carassius auratus) to cyclic hypoxia (0.8 ± 0.2 mg/l dissolved oxygen) for 9 h or more, per day, altered plasma lipid and sex steroid profiles, which in turn directly or indirectly suppressed ovarian growth and viable spermatozoa production. Hypoxia decreased total cholesterol and high density lipoprotein (HDL p < 0.05) and elevated triglycerides (TG; p < 0.05) in both sexes. Plasma steroid concentrations particularly of 17α-hydroxyprogesterone (17-HP), estradiol (E2), testosterone (T) in females, and T and 11-ketotestosterone (11-KT) in males were attenuated under diel hypoxic conditions. Intriguingly, both diel and continuous hypoxia elevated plasma E2 and vitellogenin levels in males. However, neither lipid nor steroid profiles recorded any variation in a dose-dependent manner in response to diel hypoxia. The reduced GSI, decreased number of tertiary oocytes, and motile spermatozoa in hypoxic fish clearly indicate suppression of gametogenesis. Thereby, prolonged diel cyclic hypoxia may affect valuable fishery resources and fish population structure by impairing reproductive performances and inducing estrogenic effects in males.     

Androgen secretion activity of recombinant follicle-stimulating hormone of Japanese eel, Anguilla japonica in immature and maturing eel testes

The androgen secretion activities of recombinant Japanese eel follicle-stimulating hormone (rjeFSH) were investigated in immature and maturing eel testes. The rjeFSH stimulated testosterone (T) and 11-ketotestosterone (11-KT) secretion in immature testis but not in maturing testis. This result suggests that eel FSH plays an important role through the sex steroid secretion in immature testis rather than in maturing testis.     

Production, release and olfactory detection of sex steroids by the tench (Tinca tinca L.)

The present study is concerned with pheromone communication in tench (Tinca tinca L.), establishing firstly whether males have a high olfactory sensitivity to some typical teleost sex steroids and prostaglandins; and secondly whether males and females might be able to synthesise and release some of these steroids into the water. The C₂₁ steroid, 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P) was found to give large electro-olfactogram responses with an estimated threshold of detection of 10⁻¹² M. The male tench were equally sensitive to glucuronidated 17,20β-P (10^−11.6 M) but 100 times less sensitive to sulphated 17,20β-P (11^−9.7 M). Preliminary data from cross-adaptation studies suggest that both the free and conjugated forms are detected by the same olfactory receptor(s). Male tench also had high olfactory sensitivity to prostaglandin F_2α (PGF_2α) and 15-keto PGF_2α (11^−11.5 and 10^−11.4 M). They were relatively insensitive, however, to testosterone (T), androstenedione (AD), 11-ketotestosterone (11-KT), 17β-oestradiol (E₂), 17,20β,21-trihydroxy-4-pregnen-3-one (17,20β,21-P) and 17,20α-dihydroxy-4-pregnen-3-one (17,20α-P). Radioimmunoassays were used to measure the steroids in plasma and water and all samples were processed for the measurement of free, sulphated and glucuronidated fractions. In females, free 17,20β-P, 17,20α-P, free and glucuronidated T, and AD in plasma showed the largest increases in response to injection with mammalian gonadotropin-releasing hormone analogue (GnRHa) or Ovaprim (a mixture of GnRHa and a dopamine inhibitor). Free 17,20β-P was released into the water at the greatest rate. Plasma concentrations of the two conjugated forms of 17,20β-P were also elevated 18 h after the administration of GnRHa, but not by as much as the free steroid. In males, AD and 11-KT showed the greatest increase in response to GnRHa and were moreover released into the water at a higher rate in the treated group than in the control. The data support a possible pheromonal role for free and glucuronidated 17,20β-P. This revised version was published online in July 2006 with corrections to the Cover Date.     

Plasma steroid levels and follicular development in the female staghorn damselfish Amblyglyphidodon curacao

Female staghorn damselfish were captured from coral reefs by divers and bloodsampled. Ovarian follicle type and size distribution were correlated with plasma levels of the ovarian steroids testosterone (T) and 17β-estradiol (E₂). Ovaries contained up to three clutches of vitellogenic follicles and steroid levels were maximal when all three clutches were present.