Uptake and clearance of exogenous estradiol-17β and testosterone during the early development of coho salmon (Oncorhynchus kisutch), including eggs,alevins and fry

The uptake and clearance of estradiol-17β (E₂) and testosterone (T) were examined during the initial stages of development of coho salmon (Oncorhynchus kisutch), including eyed-eggs, newly hatched alevins and first feeding fry. Radiolabeled steroids were administered through the water in tracer amounts with or without their nonradioactive form at 400 μg l⁻¹. Regardless of developmental stage, saturation levels were invariably attained earlier for T than for E₂, thus resulting in a higher incorporation of E₂. However, both steroids had similar clearance patterns. Uptake and clearance was clearly stage-dependent, being fastest in fry, intermediate in alevins and slowest in eggs. Furthermore, combined uptake and clearance patterns showed that exposure to steroid was also higher for E₂ than for T and stage-dependent, but always markedly highest in alevins. Subsequently, based on the observed elimination of the estrogen, a double immersion in E₂ at 400 μg 1⁻¹, administered 2 days apart to maximize exposure during the alevin stage, was assayed for its effect on sex reversal and found to induce the production of 100% females. We suggest that the yolk, which is present in substantial amounts during the initial stages of development in salmonids, can retain the exogenously administered liposoluble steroids, thus providing developing embryos with an extended supply of, and exposure to, these steroids well after the treatment is finished. Together, these findings help to explain the previously observed high effectiveness of sex steroids administered during early development in regulating gonadal differentiation in salmonids, the higher effectiveness of E₂ compared to T, and clarify the localization of the most sensitive period to the action of exogenous steroids at the alevin stage in the coho salmon.     

Effect of cortisol on the metabolism of 17-hydroxyprogesterone by Arctic charr and rainbow trout embryos

The effect of cortisol on the in vitro metabolism of [³H]17-hydroxyprogesterone ([³H]17OHP) was studied during embryonic development of Arctic charr (Salvelinus alpinus) and rainbow trout (Oncorhynchus mykiss). In the absence of cortisol, rainbow trout embryos metabolized [³H]17OHP largely to androstenedione (A₄) and androstenetrione (11-KA) with a minor conversion to 17,20ß-dihydroxy-4-pregnen-3-one (17,20P). In the presence of cortisol, this biosynthesis was inhibited. On the other hand, cortisol had no apparent inhibitory effect on the nature of metabolism of [³H]17OHP by Arctic charr embryos. In these embryos [³H]17OHP was metabolized mainly to 17,20P with a minor conversion to A₄ and without the formation of 11-KA that was seen in rainbow trout.When incubated in the presence of [³H]cortisol both Arctic charr and rainbow trout embryos produced 11ß-hydroxyandrostenedione (11ß-OHA) as the major metabolite, with a minor conversion to an unknown steroid. The catabolism of the cortisol by salmonid embryos may reflect the ability of the embryo to inactivate or detoxify cortisol to protect itself from the adverse effects of this biologically potent steroid hormone The study indicates the existence of species-specific differences in the nature of metabolism of [³H]17OHP and the inhibitory effect of cortisol on this metabolism.     

Serum steroid profiles in artificially maturing female Japanese eel,Anguilla japonica

To investigate whether steroid profiles in salmon pituitary homogenate (SPH)-induced artificially maturing Japanese eel, Anguilla japonica, resemble those in other, naturally maturing fishes, the daily changes in 11 steroids were analyzed for a 70-day period (average time needed to reach the maturational phase). Concentrations of most steroids were low and changed on a weekly basis, with maximum values 2–5 days after an SPH injection. Thus, pregnenolone, 17α-hydroxypregnenolone, dehydroepiandrosterone, progesterone, 17α-hydroxyprogesterone, 17α,20 β-dihydroxy-4-pregnen-3-one, androstenedione and estrone levels were barely or not detectable in serum throughout the experimental period, which is largely in keeping with what is known about oogenesis-related steroids in other fishes. In contrast, serum testosterone (T) levels were high, but fluctuated considerably with each SPH injection (about 0.3–8.3 ng/ml). The serum estradiol-17β (E₂) levels increased after SPH injections and gradually rose throughout the experiment, peaking at the end of the experimental period (about 0.2–7.8 ng/ml). Serum levels of 11-ketotestosterone (11-KT) before SPH treatment were higher (approximately 2 ng/ml) than those of the other steroid hormones (less than 0.5 ng/ml). 11-KT levels increased gradually over the experimental period, and, like E₂, levels peaked towards the end of the experimental period (about 15 ng/ml). The observed patterns for T, E₂ and 11-KT are unlike those in other fishes. Furthermore, the consistent elevations in levels of 11-KT, both before and after SPH treatment, are suggestive of an important role for this steroid in controlling oocyte growth.     

PGF<Subscript>2α</Subscript> and gonadal steroid plasma levels of successful and unsuccessful spawning <Emphasis Type="Italic">Piaractus mesopotamicus</Emphasis> (Teleostei,Characiformes) females

Gonadal steroid and prostaglandin F₂α (PGF_2α) plasma levels were evaluated in successfully (SP) and unsuccessfully ovulated (UN) female Piaractus mesopotamicus. Forty-one females were injected with crude carp pituitary extract (0.6 and 5.4 mg kg^?1 with a 24-h interval between the doses) and sampled to determine the plasma concentration of 17β-estradiol (E₂), 17α-hydroxyprogesterone (17α-OHP), 17α,20β-dihydroxy-4-pregnen-3-one (DHP), PGF_2α, and testosterone (T) after each injection (first—A1 and second—A2), and at the time of ovulation for SP and UN. Two clusters were obtained using multivariate analysis: 1—composed of all A1, all A2, and some UN; and 2—composed of all SP and some UN. Median values of E₂ plasma levels were similar between clusters; however, plasma levels of T, 17α-OHP, DHP, and PGF_2α of cluster 2 (predominantly formed by SP) were higher than those of cluster 1. Since cluster 2 contained all SP and females of this cluster presented higher levels of PGF_2α, T, 17α-OHP, and DHP, here we evidently shown in an unprecedented manner that concomitant increased levels of these substances were associated with successful ovulation in this species, but such an increase was not determinant for successful ovulation due to the presence of some UN females in the same cluster 2. These findings highlight the unexplored potential of PGF_2α to be used as an accessory tool for inducing successful ovulation for fish farming purposes.     

Investigation into the possible role of androgens in the induction of hepatic vitellogenesis in the European eel:in vivo andin vitro studies

Changes in the levels of plasma vitellogenin (Vg), estradiol (E₂) and testosterone (T) were examined following gonadal development induced by carp gonadotropin treatment (cGTH) of freshwater female yellow and silver eels (Anguilla anguilla L.). The animals received injections of cGTH (250 μg kg⁻¹ body weight) or saline vehicle three times a week, for 6 to 8 weeks. No effect of vehicle was observed. Steroidogenic activity of the ovary was stimulated by cGTH treatment as shown by the increase in circulating steroid levels in both stages. However, the responses of T, E₂ and Vg differed according to the stage of development of eels. At the yellow stage, the initial steroid plasma levels were undetectable (< 0.01 ng ml⁻¹) before treatment and ovarian steroidogenic activity was slightly stimulated following cGTH treatment; steroid levels reached their highest values after 3 weeks and 6 weeks of treatment for E₂ (0.62 ± 0.13 ng ml⁻¹ and T (0.33 ± 0.30 ng ml⁻¹), respectively. The cGTH treatment slightly increased plasma Vg levels (0.2–0.7 μg ml⁻¹ during the experiment compared with the initial values of the group. At the silver stage, the initial steroid levels were detectable (0.7 ng ml⁻¹ for E₂ and 0.1 ng ml⁻¹ for T); cGTH treatment did not significantly increase plasma E₂ level which remained at initial levels. Nevertheless, plasma T levels dramatically increased from 0.1 to 3 ng ml⁻¹ and peaked after 1 or 2 weeks of cGTH treatment; a rapid increase in plasma Vg levels occurred, reaching its highest value at 5 mg ml⁻¹ after 3 weeks of treatment. Thus, the steroid kinetic profiles in relation to the appearance of Vg in the plasma following cGTH treatment was closely related to androgen levels and there was a strong vitellogenic response induced by chronic cGTH treatment. In order to test if androgens could be implicated in the vitellogenic response, we evaluated the potencies of various androgens (testosterone and 5α-androstane-3β,17β-diol)in vivo andin vitro, associated with E₂ to induce the production of Vg.In vitro experiments showed that Vg synthesis was induced by high doses (10⁻⁶ to 10⁻⁵ M) of androgen in the eel. Tamoxifen totally inhibited the action of androgens suggesting that androgens were acting through binding to the E₂ receptor.In vivo, androgens given alone at 50 μg kg⁻¹ 3 times a week for 1 months had no significant effect on plasma Vg levels. In addition, E₂-androgen cotreatment showed that the presence of androgen did not modify the vitellogenic response induced by E₂.     

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.     

Effects of cortisol and triiodo-L-thyronine on the steroidogenic capacity of rainbow trout ovarian follicles at two stages of oocyte maturation

The in vitro basal and salmon gonadotropin (sGTH)-stimulated steroidogenic capacity of rainbow trout follicles was examined at four stages [early (EV)-, mid (MV)- and peak-vitellogenic (PV), and pre-ovulatory, post-vitellogenic (PO)] of gonadal recrudescence using radioimmunoassays (RIAs) to measure 17-estradiol (E₂) and testosterone (T) production. In addition, follicles were incubated in the presence of [³H]pregnenolone ([³H]P⁵) and the radiolabelled steroid metabolites produced were separated using high performance liquid chromatography (HPLC). Peak basal and sGTH-stimulated E₂ and T production was found in PV stage follicles and lowest in PO stage follicles, and there were marked differences in the HPLC profiles of steroid metabolites. For EV stage follicles the major metabolite eluted as a peak that co-eluted with the androstenedione (A₄) and 17-hydroxyprogesterone (17-OHP) standards. A smaller peak that co-eluted with 11-hydroxyandrostenedione (11-OHA₄) and very small peaks co-eluting with 20-dihydroprogesterone (20-DHP) and E₂ were also seen. MV and PV stage follicles produced predominantly E₂, together with a small combined A₄ + 17-DHP peak, traces of 11-OHA₄ and two peaks that did not co-elute with any of the reference standards. The PO stage follicles produced only 17, 20-dihydroxy-4-pregnene-3-one (17,20-P).In addition, the effects of cortisol and triiodothyronine (T₃) on steroidogeneis were investigated in PV and PO stage ovarian follicles. For PV stage follicles, cortisol at 100 ng ml^–1 in the incubation medium significantly suppressed both basal and sGtH- stimulated T and E₂ production relative to control treatments. T₃ at 10 ng ml^–1 in the medium had no significant effect on either basal or sGtH-stimulated T or E₂ production compared to the controls, nor did it have any beneficial effect over the suppressive effect of cortisol. PO phase follicles taken 1 to 2 weeks prior to anticipated spawning had very low E₂ and T production, and there was no effect of cortisol or T₃, alone or in combination, on E₂ or T production. For PV stage follicles incubated in the presence of [³H]P⁵, cortisol suppressed T and E₂ production, but did not block the steroid pathway at any specific level; T₃ had no apparent affect on the metabolism of [³H]P⁵. The PO stage follicles produced little or no E₂; the major metabolite was 17,20-P. Cortisol and T₃ had no apparent effect on either basal or sGtH-stimulated 17,20-P production by the follicles at this stage of maturation.     

Seasonal changes in plasma gonadal steroid concentrations and gonadal morphology of male and female tench (Tinca tinca, L.)

In order to gain a better understanding of the reproductive cycles of male and female tench (Tinca tinca), gonadosomatic index, gonad histology and plasma concentrations of estradiol‐17β (E₂), testosterone, an drostenedione, 11‐ketotestosterone (11‐KT), 17,20β, 21‐trihydroxy‐4‐pregnen‐3‐one (17,20β,21‐P), 17,20β‐dihydroxy‐4‐pregnen‐3‐one (17,20β‐P) and 17,20α‐dihydroxy‐4‐pregnen‐3‐one (17,20α‐P) were measured at the four seasons of the year, plus a further sampling coincident with the peak of spawning in early July. As expected, in both males and females, the plasma concentrations of androgens (excluding 11‐KT in females – undetectable) and C₂₁ steroids were significantly more elevated in the spring and summer (when most gonadal development took place) than in the autumn and winter. The only unexpected finding was that 17,20β‐P and 17,20β,21‐P, the steroids that are normally associated with oocyte final maturation in females and spermiation in males, were found in substantial amounts in both pre‐vitellogenic, pre‐spermatogenic and post‐spawning fish. This suggests that these steroids may have other as yet unidentified roles in this species.     

The relationship between plasma and ovarian levels of gonadal steroids in the repeat spawning marine fishesPagrus auratus (Sparidae) andChromis dispilus (Pomacentridae)

The relationship between plasma and ovarian levels of gonadal steroids was examined in two New Zealand fish species with multiple spawning cycles of differing length. Snapper (Pagrus auratus) have a daily cycle of oocyte development, ovulation and spawning, whereas demoiselles (Chromis dispilus) spawn over 2–3 days during a repeat spawning cycle of 7–9 days. Ovarian and plasma levels of the gonadal steroids 17β-estradiol (E₂), testosterone (T), 17-hydroxyprogesterone (17P) and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP) were measured in reproductively active fish captured from the wild. Ovarian levels of E₂, T and 17P changed in relation to spawning cycle and gonad stage in both snapper and demoiselles. E₂ and T levels were detectable at all times, but highest during vitellogenesis in both species. Cyclic changes of 17P occurred in both species, and levels appeared to depend on the rate of conversion of 17P to other hormones. No changes in ovarian levels of 17,20βP were detected in relation to stage of the spawning cycle in snapper; however, ovarian levels of 17,20βP were highest in demoiselles before spawning when fish undergoing final oocyte maturation predominated. Plasma levels of E₂ and T were strongly correlated with ovarian concentrations (r=0.850 and r=0.819 for E₂ and T respectively) in demoiselles but there was poor correlation between ovarian and plasma levels of 17P and 17,20βP (r=0.004 and 0.273 respectively), or between ovarian and plasma levels of E₂, T, 17P or 17,20βP of snapper (r=0.135, 0.277, 0.131 and 0.279). The poor correlation between plasma and ovarian levels of some steroid hormones suggests that plasma concentrations of steroids may not adequately reflect the reproductive status of the fish during short-term cyclic ovarian changes. It is suggested that this disparity is likely to be most marked in species with ovulatory periodicity of short duration.     

Effect of in vitro xenoestrogens on steroidogenesis in mature female fish, Chasmichthys dolichognathus

In this study, fully vitellogenic oocytes of the longchin goby (Chasmichthys dolichognathus) were exposed to in vitro xenoestrogens such as diethylstilbestrol (DES), bisphenol A (BPA) and nonylphenol (NP) at concentrations of 100 ng/ml in the presence of [³H]17α-hydroxyprogesterone as precursor. The major metabolites produced in vitro are progestogens [17α-hydroxy,20α-dihydroprogesterone (17α20αOHP) and 17α-hydroxy,20β-dihydroprogesterone (17α20βOHP)], androgens [androstenedione (A4) and testosterone (T)] and estrogens [estrone (E1) and estradiol-17β (E2)]. Comparative activities of these chemicals in oocyte steroid biosynthesis showed as follows: NP treatment resulted in clear stimulation of estrogen production, while the opposite occurred in DES treated incubation. Treatment with DES resulted in a higher synthesis of 17α20αOHP. BPA treatment increased the androgen, while estrogen synthesis was slightly inhibited. These results suggest that NP exhibited clearly estrogenic activity on the three chemicals.     

Effects of individual and binary mixtures of estrogens on male goldfish (Carassius auratus)

Adverse effects of five typical environmental estrogens, namely estrone (E₁), 17β-estradiol (E₂), 4-n-octylphenol (4-n-OP), 4-n-nonylphenol (4-n-NP) and bisphenol A (BPA) on adult male goldfish (Carassius auratus) were investigated both individually and in binary mixtures, using serum vitellogenin (VTG) induction and gonadosomatic index (GSI) as the endpoints. Doses of individual and binary mixtures of estrogens were chosen at broad ranges. Five individual estrogens induced common dose-dependent increases of serum VTG in the experimental fish when injection doses of the estrogen series were comparatively low. The levels of VTG induction in fish descended after peaked at a certain dose of the individual estrogen. Significant GSI decreases were observed in fish treated by all dose series of E₁ and E₂, and comparatively high doses of 4-n-OP, 4-n-NP and BPA when compared with that of solvent control (SC). Effects of binary mixtures of the five typical estrogens on VTG induction in male goldfish were in additive manner at low-effect doses, but divergences occurred at high dose levels, with the predicted effects by additive manner exceeding those were observed. All of GSI of fish treated by the binary mixtures were about or lower than 10^?3 %. Serious atrophy of gonads was observed in all the mixture treatment groups when compared with that of SC. These findings highlight the potential reproductive risk of fish resulted from existing mixtures of hormones in the aquatic environment, and they have important implications for environmental estrogen hazard assessment.     

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.     

In vitro follicle culture shows that progestagens are the maturation-inducing hormones (MIH) and possible regulators of the ovulation-mediating hormone PGE2 in female Eurasian perch Perca fluviatilis

In European aquaculture, Eurasian perch, Perca fluviatilis L., is perceived as one of the most highly valuable freshwater fish species and a strong candidate for the development of freshwater aquaculture. In the pursuit of improving the quality of reproduction in this domesticated species, investigating the hormones mediating the final oocyte maturation (FOM) is therefore indispensable. But, the exact nature of the maturation-inducing hormone (MIH) in Eurasian perch is unknown. To further validate the existence of a maturation-inducing activity behind potential hormonal candidates in this species, we in vitro tested a group of nine hormones: cortisol (Co), 11-deoxycortisol (11-D), corticosterone (coS), 11-deoxycorticosterone (DOC), 17α,20βdihydroxy-4-pregnen-3-one (DHP) and 17α,20β,21 trihydroxy-4-pregnen-3-one (THP), prostaglandin E₂ (PGE2), estradiol-17β (E2) and testosterone (T), in their ability to trigger FOM advancement and the production of sex steroids potentially involved in FOM. Using mature female perch, two in vitro experiments were conducted with oocytes at the start of the FOM. The follicles were incubated for 62 h in Cortland media with and without human chorionic gonadotropin (hCG). By the end of the incubation, only DHP and THP triggered the full advancement in FOM even at low doses with the effect of DHP being in vivo validated. However, the de novo productions of E2 and DHP were not shown to be regulated by either of the MIH candidates. Progestagens are hence more credible candidates as MIH than corticosteroids in Eurasian perch. Our in vitro study also revealed that both PGE2 and DHP are strongly associated with ovulation and that PGE2 might have slightly contributed to such DHP activity.

     

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.     

Correlation between oocyte development and plasma concentrations of steroids and vitellogenin in greenback flounder Rhombosolea tapirina

The development of oocytes in association with changes in plasma concentrations of vitellogenin (Vtg), 17β-estradiol (E₂) and testosterone (T) was investigated in maturing female greenback flounder Rhombosolea tapirina over the first part of a reproductive season. During vitellogenesis, increasing oocyte size was accompanied by the sustained elevation of plasma Vtg and E₂. There was a marked increase in T towards the end of vitellogenesis.     

Steroidogenic shift is a critical event for ovarian follicles to undergo final maturation

The steroidogenesis in the granulosa-thecal layers of fish ovarian follicles undergo a distinct shift from the production of estradiol-17β (E₂) to 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP, maturation-inducing hormone, MIH) prior to meiotic maturation. This review attempts to explain the underlying mechanisms of steroidogenic shift.     

Isolation and expression of rainbow trout (Oncorhynchus mykiss) ovarian cDNA encoding 3β-hydroxysteroid dehydrogenase/Δ5−4-isomerase

A distinct shift in steroidogenesis from testosterone to 17α-hydroxyprogesterone occurs in the salmonid ovarian thecal cell layers immediately prior to oocyte maturation, and is a prerequisite for the production of 17α,20β-dihydroxy-4-pregnen-3-one (maturation-inducing hormone of salmonid fishes) by granulosa cells during oocyte maturation. 17α-Hydroxylase/17,20-lyase cytochrome P-450 (P-450_17α) and 3β-hydroxysteroid dehydrogenase/Δ^5?4-isomerase (3β-HSD) are the two major steroidogenic enzymes involved in the production of 17α-hydroxyprogesterone and testosterone. Using mammalian cDNA probes, we isolated and characterized full-length cDNAs encoding these two enzymes from a rainbow trout (Oncorhynchus mykiss) ovarian thecal cell cDNA library. The cloning of 2.4-kilobase cDNA encoding P-450_17α and transient expression of this clone in nonsteroidogenic monkey kidney tumor COS-1 cells have recently been reported (Sakai et al. 1992). We have isolated a 1.4-kilobase cDNA which is hybridized to the mammalian 3β-HSD cDNAs. Expression of this cDNA in COS-1 cells led to the production of an enzyme which is capable of converting dehydroepiandrosterone to androstenedione. In this study, enzymatic activities and expression of rainbow trout ovarian P-450_17α and 3β-HSD are discussed in relation of the steroidogenic shift occurring in the ovarian follicle layers.     

Serum thyroxine,estradiol-17β, and testosterone profiles during the parr-smolt transformation of masu salmon,Oncorhynchus masou

Changes in serum thyroxine (T₄), estradiol-17β (E₂) and testosterone (T) levels during the parr-smolt transformation (smoltification) were investigated in the masu salmon (Oncorhynchus masou) in 1985 and 1987. T₄ showed a peak in levels at the early stage of smoltification and E₂ and T levels peaked almost at the same time. There were no significant differences between the concentrations of serum hormones in female and males. During smoltification, germ cells in the peri-nucleolus and spermatogonia stage were present in the ovary and testis, respectively. These results suggest that E₂ and T may be involved in smoltification in the masu salmon.