Turnover of α-, γ, and δtocopherol and distribution insubcellular and lipoprotein fractions indicate presence of an hepatic tocopherolbinding protein in Atlantic salmon (Salmo salar L.)

The purpose of this work was to study the turnover of a, andtocopherol (TOH) in Atlantic salmon (Salmo salar, L.). Fish induplicate tanks were fed a diet containing 150 mg kg^-1-TOH and 100 mg kg^-1 each of and TOHadded as tocopheryl acetates. After fillet TOH concentrations had adjustedto the dietary supplementation levels, samples were taken from fish that hadbeen deprived of feed for 100 h, and from fish that had been fed regularlyuntil sampling. The retained levels of tocopherols in plasma correspondedgrossly with their biological activities, as found in experiments withmammals (::100: 20:3). The plasma concentrationsof -, and TOH amounted to 65, 44 and 15%,respectively, in unfed compared to fed fish. Very low density lipoprotein(VLDL), appeared to contain a greater fraction of plasma -TOH than ofplasma TOH. The mitochondrial fraction of liver, but not that of darkmuscle, was highly enriched in -TOH, and less in andTOH. The concentration ratios in liver and bile indicate that, and to some extent, TOH are excreted in the bile at a higherrate than -TOH. The data fit the hypothesis that Atlantic salmonliver contains a tocopherol binding protein with higher affinity for-TOH than for the other tocopherol homologues. This appears toprevent excretion of -TOH in the bile, and stimulate incorporation of-TOH in VLDL for subsequent secretion into the blood stream. As aconsequence, -TOH is retained in the body to a greater extent than and -TOH.     

Gonadotrophin-releasing hormone agonist raises plasma concentrations of progestogens and enhances milt fluidity in male Atlantic halibut (Hippoglossus hippoglossus)

In two separate spawning seasons, spermiating male Atlantic halibut were implanted with pellets containing gonadotrophin-releasing hormone agonist (GnRHa). Males were bled repeatedly, and milt samples were collected. Blood samples were assayed for free and conjugated steroids: testosterone, 11-ketotestosterone, 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,20,21-trihydroxy-4-pregnen-3-one and steroids with a 17,20 configuration. Towards the end of the first season, pellets were implanted into three wild-caught and three hatchery-reared males. No control fish were available. The major progestogen in plasma was identified as sulphated 5-pregnane-3,17,20-triol (3,17,20-P-5-S). Concentrations of this steroid were stimulated by the GnRHa. Sulphated 17,20-P was also identified in the plasma, but at 10-fold lower concentrations than 3,17,20-P-5-S. In the middle of the second season, pellets were implanted into five hatchery-reared males; five unimplanted males were used as controls. Levels of androgens fell following GnRHa treatment, levels of progestogens rose briefly, and there was a significant increase in the fluidity of the milt. Of all the measured steroids, free and sulphated 17,20-P showed the best correlation with milt fluidity.     

Plasma profiles of fourteen ovarian steroids before,during and after ovulation in African catfish,Clarias gariepinus,determined by gas chromatography and mass spectrometry

The plasma concentrations of fourteen ovarian steroids were measured in postvitellogenic African catfish,Clarias gariepinus, which had been injected with pimozide and LHRHa. Postvitellogenesis persisted for at least four hours after pimozide and LHRHa administration. During this stage a limited rise in the plasma gonadotropin (GTH) level was accompanied by an increase in the testosterone concentration. The estradiol level was high and remained high except for a passing drop during the stage of germinal vesicle migration. At the stage of germinal vesicle migration a strong increase in the plasma GTH level coincided with a maximum in the testosterone concentration and a concomitant increase in the levels of 17,20-dihydroxy-4-pregnen-3-one and of five 5-reduced pregnanes. During germinal vesicle breakdown the GTH concentration remained high, the plasma level of 17-hydroxyprogesterone tended to increase, and the levels of 5-pregnane-3, 17-diol-20-one, 5-pregnane-3,17,20-triol and 5-pregnane-3,17,20-triol reached a maximum. At pre-ovulation the GTH concentration did not change, and peak levels were reached of 17,20-dihydroxy-4-pregnen-3-one and 5-pregnane-3,6,17-triol-20-one. Shortly after ovulation the GTH concentration slightly decreased together with a sharp decline in the concentrations of 17,20-dihydroxy-4-pregnen-3-one and the 5-reduced steroids, with the exception of 5-pregnane-3,17,20-triol, 5-pregnane-3,6,17,20-tetrol and 5-dihydrotestosterone. The plasma concentrations of androstenedione, estrone, etiocholanolone and 5-androstane-3,17-diol showed marginal fluctuations during oocyte maturation and ovulation. Apart from 17,20-dihydroxy-4-pregnen-3-one, the 5-reduced pregnanes might be candidates for the function of oocyte maturation inducing hormone inC. gariepinus.     

Synthesis and regulation of 17α-hydroxy-20β-dihydroprogesterone in immature males of Oncorhynchus mykiss

Three experimental approaches were chosen to study the question if the progestin 17-hydroxy-20-dihydroprogesterone (1720OHP) is synthesised in testes of young Oncorhynchus mykiss, in which the absence of spermatozoa was verified histologically: first, in order to detect 20-hydroxysteroid dehydrogenase activity (20HSD), testes homogenates were incubated with ³H-labeled 17OHP.Metabolites were analysed by TLC, HPLC, and repeated crystallization to constant isotope ratios. One of the metabolites was identified as 1720OHP-³H, indicating that already immature testes contain 20HSD activity and are able to produce 20-reduced steroids. Second, 1720OHP was quantified by radioimmunoassay in incubates of testes fragments. The sensitivity of the gonads to gonadotropin II (GtH II) became evident when comparing incubations in the absence and presence of GtH II. Third, plasma levels of 1720OHP were significantly higher in animals injected with partially purified salmon gonadotropin, compared to controls. Thus, for the first time, it could be shown that 20HSD is present in testicular cells other than spermatozoa. Furthermore, 1720OHP is indeed secreted at a very early stage of testicular development; 1720OHP secretion is also responsive to GtH II. Future studies will have to show if the functions of this progestin include the stimulation of spermatogenesis.     

Effects of steroid hormones on in vitro oocyte maturation in white sturgeon (Acipenser transmontanus)

The in vitro effects of several steroids on the maturation of intact white sturgeon (Acipenser transmontanus) ovarian follicles were investigated. At the highest concentration (1024 ng ml^–1 for the C21 steroids and 1139 ng ml^–1 for the C19 steroids), all of the C21 steroids tested, progesterone (P4), 17-hydroxyprogesterone (17OHP), 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,(20,21-trihydroxy-4-pregnen-3-one 20-S), 11-deoxycortisol (S) and cortisol (F), as well as testosterone (T) induced germinal vesicle breakdown (GVBD) at 14 and 22 h. At 6 h, only P4 and 17,20-P induced maturation at the highest concentration (1024 ng ml^–1). At 14 and 22 h, 11-deoxycortisol was the most potent steroid inducer of GVBD followed by P4, 17OHP, 17,20-P, and 20-S. The steroid 11-hydroxytestosterone (11OHT) was completely ineffective at all concentrations and exposure times. The C21 steroids induced oocyte maturation at concentrations ranging from 4 to 1024 ng ml^–1, whereas T induced GVBD at 225 to 1139 ng ml^–1. Calculation of the mean effective concentration that induced 50% GVBD (EC50) from the 22 h incubations revealed the following order of potencies: S > P4 > 17OHP > 17,20-P > 20-S >> F > T. These bioassay results, together with previous findings on the endogenous production of steroids by ovarian follicles from gonadotropin-primed females, indicate that more than one steroid has a biological role in the resumption of meiosis in sturgeon oocytes and provides empirical evidence for P4, 17OHP, S, 20-S, and 17,20-P as maturation-inducing steroids in white sturgeon.     

Oocyte maturation inClarias batrachus. III. Purification and characterization of maturation-inducing steroid

Maturation-inducing steroid (MIS) in the Indian female catfish,Clarias batrachus, was purified and characterized from the incubation medium in which fully grown but immature folliculated oocytes were incubated with salmon gonadotropin (SG-G100) for 36 h. Maturation-inducing (MI) activity of residues obtained at various steps of extraction and purification was assessed byin vitro germinal vesicle breakdown (GVBD) assay using folliculated oocytes ofC. batrachus. The post incubation medium was extracted with diethyl ether. The ether phase was partitioned using 50% methanol plus n-hexane. The methanol phase which had MI activity was fractionated into 7 fractions using reverse-phase high-performance liquid chromatography. Of these 7 fractions, fraction 3 was found to be active in having MI ability and identified as 17 ,20-dihydroxy-4-pregnen-3-one (17,20-diOHprog). The authenticity of 17,20-diOHprog as the major follicular mediator of gonadotropin-induced oocyte maturation was further confirmed by thin-layer chromatography (TLC) in which fraction 3 was run along with authentic 17,20-diOHprog standard. This investigation gives a direct evidence that 17,20-diOHprog is the major naturally occurring MIS in Indian female catfish,C. batrachus.     

Brackishwater carp culture in potentially waterlogged areas using animal wastes as pond fertilizers

In order to assess the possibilities of utilizing drainage effluents (salinity range 5.0–12.5), fish culture experiments were carried out. Experiments on polyculture using cow dung (24 000 kg ha^–1 y^–1) as pond fertilizer were conducted at five different salinity levels (0.3–8.5). Studies have revealed that carp perform well in salinities up to 7.5 and reasonably high fish production has been obtained. Even though the ponds had a high trophic status, higher salinities ( > 7.5) appear to repress fish growth probably due to low dissolved oxygen (DO), high BOD and high NH₄-N. Experiments on monoculture of common carp (Cyprinus carpio) conducted at two different salinity levels (0.3–0.9 and 6.0–7.0) using four different organic fertilizers (cow dung at 24 000 kg and 20 000 kg, poultry at 1500 kg, duck at 6000 kg and sheep/goat at 1500 kg ha^–1 y^–1) have revealed the highest fish growth to be in poultry-treated ponds, followed in decreasing order by duck and sheep/goat wastes. Similar trends in fish production were observed both in fresh- and salt-water ponds. However, fish production was lower in ponds having higher salinities ( > 7.5). Nevertheless, these studies indicated that inland saline waters can be utilized for fish culture. With minor modifications in the existing technology of fish culture in stagnant freshwater fish ponds, animal wastes could be used to fertilize brackishwater fish ponds.     

Steroidogenesis by ovaries and testes of the European catfish,the wels (Silurus glanis), in vitro

Testosterone, 3,17-dihydroxy-5-pregnen-20-one, 17,20-dihydroxy-4-pregnen-3-one (17,20P) and 5-pregnane-3,17,20-triol were identified as the major metabolites of [³H] 17-hydroxyprogesterone in ovarian incubations of the European catfish Silurus glanis. 17,20P and the reduced triol were present only in ovaries from fish primed with carp hypophysial homogenate (chh) while testosterone yields were significantly higher in controls than in treated fish. 11-Ketotestosterone, 11-hydroxytestosterone and 17,20-dihydroxy-4-pregnen-3-one (17,20P) were identified as the major metabolites of [³H]17-hydroxyprogesterone in in vitro incubations of testes of a spermiating catfish. There was no significant production of conjugates or other water soluble metabolites by either sex. The stimulation of plasma 17,20P, 17,20P and 11-hydroxytestosterone by chh in primed but not control males suggests that the role of these steroids in spermiation should be further examined.     

Seasonal,reproductive, and nutritional influences on muscle “buffering capacity” in yellow perch (Perca flavescens)

Effective non-bicarbonate buffering capacity (or buffer value) was measured in white muscle of yellow perch (Perca flavescens) by titrations with mineral acid and base in a carbon-dioxide free, closed system. Yellow perch were collected at three month intervals throughout 1983 from an acidic lake (pH 4.6) and two alkaline lakes (pH 7.8) in northern Wisconsin. Buffering capacity was also determined for white muscle of perch kept in the laboratory under different regimes of temperature and ration. The mean buffering capacity of white muscle from yellow perch taken directly from natural environments ranged from 40.7 ± 3.1 (SD) slykes in March of 1983 to 53.7 ± 2.8 (SD) slykes in July of that year. These changes in buffering capacity were strongly correlated with water temperature. Egg production and thirty-day laboratory starvation produced significant decreases in buffering capacity and increases in the water content of yellow perch muscle. Fed perch in the laboratory had a temperature dependent buffering capacity similar to field caught fish. Buffering capacity of white muscle did not differ between yellow perch from acidic and alkaline lakes. Investigators using buffering capacity as a gauge of species differences in metabolic potential, should be wary of seasonal and reproductive factors that might alter their conclusions.     

Effects of acute temperature decreases on turbot fry and juveniles

Turbot fry (10–20 mm) and juveniles (85–110 mm) were transferred directly from 16.0–16.5 C to 1.0 C, 2.5 C, 5.5 C or 8.0 C seawater. The fry were more sensitive to cold water than juveniles. The fry survived for 1 week at 8.0 C but not at 5.5 C, whereas juveniles survived at 5.5 C but not at 2.5 C. Transfer of juveniles to 1.0 C and 2.5 C seawater caused a high mortality, a marked increase in plasma Cl^- concentration, decrease in muscle water content, and hyperglycaemia. Acclimation to 5.5 C (juveniles) or 8.0 C (fry and juveniles) markedly reduced the sensitivity to 1.0 C exposure.     

Regulation of oocyte maturation in the rainbow trout,Salmo gairdneri: role of cyclic AMP in the mechanism of action of the maturation inducing steroid (MIS), 17α-hydroxy, 20β-dihydroprogesterone

In fish, oocyte maturation (resumption of meiosis after completion of vitellogenesis and before ovulation) is triggered by maturation inducing steroids (MIS) which generally appear to be secreted in the ovary in response to stimulation by a pituitary maturational gonadotropin. Converging data from different laboratories show that 17-hydroxy, 20-dihydroprogesterone (17, 20-OH-P) is the principal MIS in salmonoids; but clear identification remains to be done in other taxonomic groups.The experiments reported here in the rainbow troutSalmo gairdneri examine the possible involvement of oocyte cAMP on the mechanism of MIS action. The action of 17, 20-OH-P, on germinal vesicle breakdown (GVBD) in oocytes incubatedin vitro within the follicle, was inhibited by various substances expected to elevate the intraoocyte concentrations of cAMP: cAMP ( 1 mM) or dibutyril cAMP ( 2 mM), phosphodiesterase inhibitors such as theophylline ( 0.2 mM) or 3-isobutyl-1 methylxanthine (IBMX 0.1 mM), adenylate cyclase activators such as cholera toxin (> 100 nM) or forskolin ( 0.03 mM). In fact, the combined action of IBMX (1 mM) and forskolin (0.01 or 0.05 mM)in vitro was to promote accumulation of intraoocyte cAMP within 1 to 5 hours. Oocyte cAMP concentrations exhibited a large variability between different females, depending on the stage of oocyte development; a significant positive correlation between oocyte cAMP concentration and the follicular weight, and a significant negative correlation between oocyte cAMP concentration and the median efficient dose of 17, 20-OH-P for induction of GVBD, were observed. Finally, when intrafollicular oocytes were incubatedin vitro, the addition of a maturation-inducing concentration of 17, 20-OH-P (3×10^–6M) induced a significant decrease of oocyte cAMP within the first 10 hours of incubation. These results show that cAMP appears to play a central role in the regulation of oocyte sensitivity to 17, 20-OH-P and in the intraoocyte mechanisms leading to GVBD in trout.These data are discussed together with the few indications available in fish concerning the mechanism of MIS action which can be compared to some extent with the amphibian model.     

Binding characteristics and regulation of the 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S) receptor on testicular and sperm plasma membranes of spotted seatrout (Cynoscion nebulosus)

The binding characteristics of 17,20,21-trihydroxy-4-pregnen-3-one (20-S) to plasma membranes prepared from the testes and sperm of spotted seatrout (Cynoscion nebulosus) were investigated using a filtration method to retain the bound 20-S. A single class of high affinity (K_d = 17.9 nM), low capacity (B_max = 0.072 nM g^-1 testes) binding sites was identified by saturation and Scatchard analyses on testicular membranes of spermiating spotted seatrout. A corresponding receptor (K_d = 22.17 nM, B_max = 0.00261 nM ml^-1 milt) was also detected in spermatozoan membrane preparations. The rates of 20-S association and dissociation were rapid, both had T_halfs of less than 1 min. Competition studies indicated that the receptor was highly specific for 20-S. 17,20-dihydroxy-4-pregnen-3-one, which had the highest affinity of the other steroids tested, had a relative binding affinity (RBA) of 14.3%. Progesterone, 11-deoxycortisol and testosterone competed with an order of magnitude less affinity (RBA's of 7.4, 1.8 and 1.1%, respectively). Estradiol displayed low affinity for the receptor (RBA = 0.4%) and cortisol did not cause any displacement at 1000-fold excess concentration. Specific 20-S receptor binding was detected in plasma membranes from testes of both spermiating and non-spermiating seatrout and on spermatozoa. Prolonged incubation of testicular fragments from a spermiating fish with gonadotropin (15 IU ml^-1 human chorionic gonadotropin) or forskolin (10 µM) caused a 2–3 fold increase in membrane receptor binding. Previous studies have shown that gonadotropin-induced upregulation of the 20-S plasma membrane receptor in seatrout ovaries is required for the oocytes to become responsive to 20-S and undergo final maturation. The existence of a 20-S membrane receptor on sperm and its upregulation in the testes by gonadotropin raises the possibility that final maturation of spermatozoa in male seatrout may be regulated by a similar mechanism.     

The nature of hematological response in fish

Hematological status was examined in rainbow trout,Oncorhynchus mykiss, held for 3–4 weeks under temperature, photoperiod and P_{O 2} conditions approximating those of their winter, spring and summer habitats. The most striking change observed was in red cell population composition. In winter fish mature cells were predominant; juvenile and developing erythrocytes characterized spring and summer animals. Hemoglobin, hematocrit and both mean erythrocytic volume and hemoglobin were modestly lower in spring and summer than in winter fish. Red cell numbers were not significantly affected. These observations suggest that avoidance of viscosity-based increases in circulatory work cost is more advantageous than elevation of blood O₂-carrying capacity. Although hemoglobin isomorph profiles were significantly altered, there is little evidence that such changes are of critical adaptive importance. Given presumed age-based reduction in gas transport effectiveness, the replacement of mature and senescent cells by more metabolically-competent juvenile cells appears to be the pivotal event in hematological response. Leucocyte counts were significantly elevated in spring and summer as compared to winter fish. Lymphocyte/heterophil ratios declined from 8.27 in winter fish to 3.13 in summer trout. Thrombocyte, monocyte, eosinophil and basophil abundances were little changed.     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.     

Circulating levels and in vitro production of two maturation-inducing hormones in teleost: 17α,20β-dihydroxy-4-pregnen-3-one and 17α,20β,21-trihydroxy-4-pregnen-3-one, in a daily spawning wrasse, Pseudolabrus japonicus

The female bambooleaf wrasse, Pseudolabrus japonicus, spawns daily during the spawning season, and exhibits a diurnal rhythm of ovarian development. In the present study, we have investigated: (1) circulating levels of 17a, 20-dihydroxy-4-pregnen- 17,20-P) and 17,20,21-trihydroxy-4-pregnen-3-one (20-S) in females sampled at different times of the day during spawning season in captivity, and (2) in vitro production of 17,20-P and 20-S by follicle-enclosed oocytes at seven different develo tal stages. In addition, we developed a microtiter plate enzyme-linked immunosorbent assay (ELISA) for 17,20-P. Serum levels of 17,20-P and 20-S showed similar diurnal changes; substantial increases in these levels occurred around the time of germinal vesicle breakdown (GVBD). In vitro experiments showed that massive production of 17,20-P and 20-S occurred in follicles collected just before or during GVBD. Further, acute decreases in 17,20-P and 20-S production were found in the ovarian follicles just prior to ovulation, suggesting inactivation of the maturation-inducing hormone (MIH). These results, taken together with our previous data on the occurrence of GVBD in vitro, suggest a role for both 17,20-P and 20b-S as MIHs in the bambooleaf wrasse.     

Gonadotropins I and II do not stimulate thein vitro secretion of 17α,20β-dihydroxy-4-pregnen-3-one 20-sulphate by rainbow trout gonads during final sexual maturation

Thein vitro secretion of 17,20-dihydroxy-4-pregnen-3-one 20-sulphate (17,20-P-sulphate) and the free steroid 17,20-dihydroxy-4-pregnen-3-one (17,20-P), by rainbow trout (Oncorhynchus mykiss) gonads, in response to gonadotropin (GTH) I and GTH II, were studied during the final stages of sexual maturation. Substantial amounts of 17,20-P-sulphate were produced, by both mature ovaries and testes, indicating considerable 20-hydroxysteroid sulphotransferase (20-HST) activity within these tissues. In the post-ovulatory ovary the level of 17,20-P-sulphate (36.6 ng ml^–1) greatly exceeded that of 17,20-P (8.59 ng ml^–1). The amount of 17,20-P-sulphate produced in incubations of both mature ovary and testes was unaffected by either GTH I or GTH II treatment at physiological concentrations up to 100 ng ml^–1. Similarly, incubations of maturing ovary and testes, treated with GTH I or GTH II, in the presence of added 17,20-P at 100 ng ml^–1 of medium, produced levels of 17,20-P-sulphate that were similar to those of the controls. In incubations of mature ovarian follicles at the stages of germinal vesicle breakdown and preovulation, both GTHs significantly stimulated secretion of 17,20-P, although GTH II was always more potent than GTH I. GTH II significantly elevated the levels of 17,20-P in testicular incubations from mature males more than 4-fold relative to GTH I and controls, which did not differ from one another.In conclusion, 20-HST, the enzyme responsible for the sulphate conjugation of 17,20-P, was found to be active in the ovaries and testes of rainbow troutin vitro. However, the levels of this enzyme do not appear to be regulated by either GTH I or GTH II.     

Smoltification induced by a ‘skeleton’ photoperiod in underyearling coho salmon (Oncorhynchus kisutch)

Underyearling coho salmon fry were subjected to three initial photoperiod treatments (6L18D, 10L14D, 14L10D) for two months and subsequently to three final treatments (16L8D, 9L6D1L8D, 10L14D) in a factorial design. Growth rates and seawater adaptability were monitored regularly. The groups that were exposed initially to 6L18D or 10L14D and then to 16L8D grew faster and had lower plasma sodium ion levels after seawater challenge tests than any of the other groups. Fish which were initially exposed to 6 L or 10 L daylength and then to a 9L6D1L8D skeleton photoperiod, showed a slightly lower growth rate and seawater adaptability than those given the corresponding complete 16L8D photoperiod. However fish maintained on skeleton photoperiods had significantly greater growth rates and seawater adaptability than those kept on the 10L14D photoperiod. This indicates that it is not the accumulated number of hours of exposure to light that initiates smolting, but rather the time during the day when light is experienced. Fish exposed initially to 14L10D showed little or no response to subsequent changes in photoperiod, suggesting that responsiveness to inductive photoperiods depends on the initial photoperiod treatment.     

Progestogen metabolism by ovaries of the roach (Rutilus rutilus L.) and the rudd (Scardinius erythrophthalmus L.)

Roach ovaries converted 17-hydroxyprogesterone to 17,20-dihydroxy-4-pregnen-3-one (17,20P) and to glucuronides of testosterone and 17,20P. Small amounts of 5-pregnane-3- and -3, 17, 20-triols, 7-hydroxy-5-reduced metabolites and 17,20-dihydroxy-4-pregnen-3-one (17,20P) were also formed. Rudd ovaries converted this substrate mainly to 17,20P, 5-pregnane-3- and -3,17,20-triols, 17,20-dihydroxy-5-pregnan-3-one and testosterone glucuronide. The main metabolites of progesterone with both species were 17,20P, 5-pregnane-3,17,20-triol and 7-hydroxy-5-reduced steroids. Rudd ovaries formed, in addition, 17,20-dihydroxy-5-pregnan-3-one from progesterone. The pattern of metabolites was markedly altered when the concentration of substrate was increased from 42ng to 1 µg or 100 µg. At the highest concentration, glucuronides and polar steroids were not detectable, while at low concentrations they accounted for over 50% of the metabolites. 20-Hydroxysteroid dehydrogenase was shown to have a very high capacity, producing 21–47 µg 17,20P from 100 µg 17-hydroxyprogesterone substrate with 200 mg ovarian tissue in 5h.     

Modulation of glycoprotein hormone α- and gonadotropin IIβ-subunit mRNA levels in the pituitary gland of mature male African catfish, Clarias gariepinus

The cDNAs encoding the glycoprotein hormone -subunit (GP) and the gonadotropin II-subunit (GTH II) were cloned from the pituitary gland of the African catfish, Clarias gariepinus. Using RNase protection analysis, we studied the steady-state mRNA levels of GP as well as GTH II in the pituitary gland of adult male catfish. Castration of adult male catfish resulted in a significant decrease of GTH II mRNA levels, whereas there was no change in the GP mRNA levels. Treatment of intact males with a single dose of 11-ketotestosterone (11-KT) resulted in dose-dependent increases in mRNA levels of both GP and GTH II. We conclude that 11-KT, a prominent, non-aromatizable teleost androgen, has a stimulatory effect on the pituitary mRNA levels of GP and GTH II of adult male fish.     

Australia‘s Lake Eacham rainbow fish: lessons and outlook

Recent DNA comparisons confirm that the endangeredLake Eacham rainbow fish (Melanotaenia eachamensis Allen andCross, 1982) is a separate species, distinct from othermelanotaeniids in the area. Although the large populationformerly in Lake Eacham has not been re-established, other smallnatural populations of M. eachamensis have been identified fromLake Euramoo and Dirran Creek.Commencing with 23 captive individuals (3 14 6 I), representing three original stocks, Project Eachamensis (initiated by the Australia New Guinea Fishes Association in July 1987) has stimulated and actively encouraged the establishment of new captive populations. Approximately 15 captive breeding populations are now reported in eastern Australia -- numbers of individuals are estimated to exceed 1000; additional Bowman and Tappin populations are being maintained in North America and Europe.A recent review of this conservation programme (involving individual aquarists, hobbyist organizations and scientists as well as the aquarium industry and government) has now recommended: an alteration to the conservati on status category from endangered to vulnerable; maintenance of existing captive populations; formulation of a comprehensive recovery plan with the long-term objective of re-establishing and increasing natural populations; and encouraging public education on the risks of translocation.