Estrogen and glucocorticoid receptor agonists and antagonists in oocytes modulate the pattern of expression of genes that encode nuclear receptor proteins in very early stage rainbow trout (Oncorhynchus mykiss) embryos.

Previous studies show that changes in estrogen (ER) and glucocorticoid receptor (GR) function in rainbow trout (Oncorhynchus mykiss) oocytes modulate the growth performance phenotype of embryo and juvenile progeny; the present study was undertaken to determine whether this altered growth performance is associated with changes in the expression of several growth-related genes in early-stage embryos. Unfertilized oocytes were incubated in the presence of various combinations of GR and ER agonists and antagonists; the oocytes were then fertilized and the expression of genes that encode for six nuclear receptor superfamily (NRS) proteins (GR1, GR2, ERα, ERβ, TRα, and TRβ) and the two IGF peptides (IGF1 and IGF2) were measured in the 7-, 13-, and 26-dpf embryos. By day 26 of embryogenesis, the expression of the six NRS-related genes of interest and that of igf2 were significantly enhanced in embryos reared from ER agonist- or ER antagonist-treated oocytes, regardless of whether the GR agonist, cortisol, was also included in the initial oocyte incubation medium.

Cortisol and dexamethasone increase the in vitro multiplication of the haemoflagellate, Cryptobia salmositica, possibly by interaction with a glucocorticoid receptor-like protein.

Cryptobia salmositica is a pathogenic haemoflagellate of Pacific salmon, Oncorhynchus spp., on the west coast of North America. The in vitro multiplication of the parasite was significantly enhanced by the addition of cortisol (within a range consistent with physiological levels in salmonid fishes; 10-50 ng ml(-1)) to the culture medium (MEM supplemented with FBS).

The in vitro metabolism of cortisol by ovarian follicles of rainbow trout (Oncorhynchus mykiss): comparison with ovulated oocytes and pre-hatch embryos.

Mid-vitellogenic stage rainbow trout (Oncorhynchus mykiss) ovarian follicles (both intact and yolk free (YF)), ovulated oocytes and embryos were co-incubated with [2,4,6,7-(3)H]cortisol for 18 h to determine the degree and nature of the metabolism and biotransformation of the glucocorticoid. There was evidence of the conversion of cortisol to the less biologically potent glucocorticoid, cortisone, and the formation of glucocorticoid sulphates (both cortisol and cortisone) for all cell and tissue samples, suggesting the presence of 11β-hydroxysteroid dehydrogenase (11β-HSD) and glucocorticoid sulphotransferase (GST) activity at all stages; however, GST activity was particularly marked in both intact and YF ovarian follicles, suggesting an important role of follicles in limiting the exposure of oocyte to maternal cortisol. As there was no evidence of 11β-HSD or GST activity in ovarian fluid, the findings affirm that ovarian follicles (probably the thecal and granulosa cells) provide a barrier against the transfer of cortisol to the oocytes by forming sulphated steroids, whereas ovulated oocytes and early embryos have a more limited capacity to either metabolize or conjugate cortisol and are therefore more vulnerable at the post-ovulatory and early embryonic stages to increases in exposure to the glucocorticoid.

Glucocorticoid receptor activation following elevated oocyte cortisol content is associated with zygote activation, early embryo cell division, and IGF system gene responses in rainbow trout.

Increased in ovo cortisol content of rainbow trout oocytes from ~3·5 to ~5·0 ng.oocyte(-1) before fertilization enhances the growth of embryos and juveniles and changes the long-term expression pattern of IGF-related genes. This study used embryos reared from oocytes enriched with cortisol and the glucocorticoid receptor (GR) antagonist, RU486, to determine whether the growth-promoting actions of cortisol involve GR protein activation and modulation of gr expression.

The influence of oocyte cortisol on the early ontogeny of intelectin and TLR-5, and changes in lysozyme activity in rainbow trout (Oncorhynchus mykiss) embryos.

The ontogeny of lysozyme activity, intelectin, TLR-5M and TLR-5S gene expression and intelectin localization was examined in rainbow trout (Oncorhynchus mykiss) reared from oocytes immersed for 3h prior to fertilization in either ovarian fluid alone (CC) or cortisol-enriched ovarian fluid at either 100 ng mL(-1) (C1) or 1000 ng mL(-1) (C2) [final oocyte cortisol concentrations were ~3, ~5, and ~7.5 ng oocyte(-1) for the CC, C1 and C2 treatment groups, respectively]. Lysozyme activity was elevated in the cortisol-treated groups from the zygote until 13-days post fertilization (dpf), but was not affected at 21-dpf.

The actions of in ovo cortisol on egg fertility, embryo development and the expression of growth-related genes in rainbow trout embryos, and the growth performance of juveniles.

Rainbow trout (Oncorhynchus mykiss) oocytes were incubated for 3 hr in ovarian fluid alone (CC), or cortisol-enriched ovarian fluid [100 or 1,000 ng ml(-1) (CL and CH, respectively)], after which they were fertilized; the growth and development of the embryos reared from these oocytes was monitored until first feed, and the juveniles were monitored for 9 months. The hatching rates of the CH group were significantly reduced, but the overall survival as measured at 40-week post-fertilization was similar in the three treatment groups. In addition, significant apparently biphasic changes relative to the CC group were found in the expression of some key growth-related genes in the CL and CH treatment groups, particularly IGF-1, IGF-2, GH1, GH2, GH receptors, and thyroid hormone receptors (TRα and TRβ).

Bisphenol A in oocytes leads to growth suppression and altered stress performance in juvenile rainbow trout.

Background: Bisphenol A (BPA), used in the manufacture of plastics, is ubiquitously distributed in the aquatic environment. However, the effect of maternal transfer of these xenobiotics on embryonic development and growth is poorly understood in fish. We tested the hypothesis that BPA in eggs, mimicking maternal transfer, impact development, growth and stress performance in juveniles of rainbow trout (Oncorhynchus mykiss).

Expression profiles of growth-related genes during the very early development of rainbow trout embryos reared at two incubation temperatures.

Quantitative RT-PCR was used to determine the profiles of expression of 10 growth- or development-related genes in rainbow trout (Oncorhynchus mykiss) embryos prior to the formation of the somatotropic (ST) axis (pituitary somatotrops and liver); embryos were sampled immediately after fertilization and water-hardening (t(0)), 1-h post-fertilization, and 1-, 2-, 5-, 7-, 10- and 13-days post-fertilization (dpf); expression profiles were examined in embryos reared at two temperatures (6.0 and 8.5 degrees C), which had different developmental rates.

Effect of glutamate and somatostatin-14 on basal and cAMP-stimulated steroidogenesis by rainbow trout (Oncorhynchus mykiss) ovarian follicles, in vitro.

The effects of glutamate and somatostatin-14 (SRIF) on the in vitro basal and cAMP-stimulated steroid production of mid-vitellogenic rainbow trout (Oncorhynchus mykiss) ovarian follicles were investigated. cAMP-stimulation was achieved by the addition of the adenylyl cyclase activator, forskolin (FS), or a membrane permeate cAMP agonist, 8-bromo-cAMP (BA), to the incubation medium. Testosterone (T) and 17beta-estradiol (E(2)) secretion was measured using radioimmunoassay.

Ah Receptor-mediated impairment of interrenal steroidogenesis involves StAR protein and P450scc gene attenuation in rainbow trout.

The objective of the study was to investigate the impact of aryl hydrocarbon receptor (AhR) activation on interrenal steroidogenesis in rainbow trout. To this end, fish were fed AhR agonist (beta-naphthoflavone (BNF): 10 mg/kg body mass/day) and antagonist (alpha-naphthoflavone (ANF): 10 mg/kg body mass/day) either singly or in combination (ABNF) for 5 days to elucidate the mechanisms involved in AhR-mediated depression of cortisol production. Liver AhR protein expression was significantly elevated only with ABNF, but not with BNF and ANF compared to the control group.

Effect of glutamate on basal steroidogenesis by ovarian follicles of rainbow trout (Oncorhynchus mykiss).

The effects of glutamate on the in vitro basal steroid production of three maturational stages of rainbow trout (Oncorhynchus mykiss) ovarian follicles were investigated. Radioimmunoassays were used to measure the rates of synthesis of testosterone (T) and 17-estradiol (E2). High performance liquid chromatography (HPLC) was used to examine the steroid metabolites produced from a tritium labeled precursor, pregnenolone (P5).

Trafficking of L-triiodothyronine between ovarian fluid and oocytes of rainbow trout (Oncorhynchus mykiss).

The study examines the dynamics of thyroid hormone (TH) trafficking between rainbow trout (Oncorhynchus mykiss) oocytes and ovarian fluid (OF) to explore the processes involved in the transfer of hormone to the oocytes. We also examined the effects of enhancing oocyte T(3) content and subsequent embryo survival. Oocytes incubated in OF alone had significant losses of THs within 12 h, whereas the T(3) content of oocytes retained in T(3)-enriched OF (10 and 100 microg ml(-1)) was significantly elevated in a dose-dependant manner within 3 h.

Effect of alloxan and insulin immunoneutralization on circulating thyroid hormone levels in larval landlocked sea lampreys, petromyzon marinus.

The effects of alloxan, an insulin (INS)-secreting cell toxin, and INS immunoneutralization on circulating levels of thyroid hormones (thyroxine, T(4); triiodothyronine, T(3)) were examined in larval landlocked sea lampreys, Petromyzon marinus. Animals were injected intraperitoneally with either (Experiment 1) saline (0.6%) or alloxan (20 or 200 microg/g body weight), or with (Experiment 2) normal rabbit serum or anti-lamprey INS.