Post-ovulatory egg viability in the snapper Pagrus auratus (Sparidae)

1997 ◽  
Vol 48 (5) ◽  
pp. 385 ◽  
Author(s):  
A. C. Hobby ◽  
N. W. Pankhurst

The role of gonadal steroids in maintaining post-ovulatory egg viability was investigated in the snapper (Pagrus auratus), a daily-repeat-spawning teleost. Ovulated eggs were held in vitro in L15 medium, or in L15 medium supplemented with gonadal steroids or gonadotrophin, to investigate any direct effects of reproductive hormones on post-ovulatory egg viability. The viability of eggs retained in the oviduct decreased with increasing time after ovulation, to give fertilization of below 50% after 6 h. The viability of eggs held in L15 medium was at least as good at that of eggs held in vivo. Storage of eggs in L15 medium supplemented with the reproductive hormones testosterone, 17α-hydroxyprogesterone, 17α,20β-dihydroxy-4-pregnen-3-one and human chorionic gonadotrophin did not improve or reduce viability relative to controls, suggesting that the hormones investigated have no direct effect on post-ovulatory egg viability.

1995 ◽  
Vol 46 (2) ◽  
pp. 467 ◽  
Author(s):  
AR Ventling ◽  
NW Pankhurst

This study investigated the role of steroids and gonadotrophin in final oocyte maturation (FOM) in the New Zealand snapper (Pagrus auratus) by means of in vitro techniques. Ovarian fragments containing vitellogenic oocytes were incubated for up to 48 h with a range of doses of gonadal steroids andlor human chorionic gonadotrophin (hCG). The ability of ovarian tissue to synthesize putative maturational steroids was assessed by measuring production of 17α,20β-dihydroxy-4-pregnen- 3-one (17,20βP) by ovarian fragments incubated with hCG and 17α-hydroxyprogesterone (17P). 17,20βP was consistently the most effective maturational steroid, inducing FOM at doses as low as 1 ng mI-1. The next most potent steroids were 20β-hydroxylated progestins and 17P, which were effective at relatively high doses (100 ng mI-1) only. The single androgen tested (testosterone) was least effective at inducing FOM. Oocytes were less responsive to hCG alone than to steroids, but the combination of hCG and steroids enhanced responsiveness in some incubations. Oocytes produced 17,20βP in response to treatment with 17P, and the response was augmented by co-treatment with hCG. HCG alone was ineffective at stimulating production of 17,20βP. The results of this study are consistent with reports that 17,20βP is generally the most potent steroid for the induction of FOM in teleosts.


2009 ◽  
Vol 2009 ◽  
pp. 1-13 ◽  
Author(s):  
M. K. Gill-Sharma

In the last 20 years, a pituitary-hypothalamus tissue culture system with intact neural and portal connections has been developed in our lab and used to understand the feedback mechanisms that regulate the secretions of adenohypophyseal hormones and fertility of male rats. In the last decade, several in vivo rat models have also been developed in our lab with a view to substantiate the in vitro findings, in order to delineate the role of pituitary hormones in the regulation of fertility of male rats. These studies have relied on both surgical and pharmacological interventions to modulate the secretions of gonadotropins and testosterone. The interrelationship between the circadian release of reproductive hormones has also been ascertained in normal men. Our studies suggest that testosterone regulates the secretion of prolactin through a long feedback mechanism, which appears to have been conserved from rats to humans. These studies have filled in a major lacuna pertaining to the role of prolactin in male reproductive physiology by demonstrating the interdependence between testosterone and prolactin. Systemic levels of prolactin play a deterministic role in the mechanism of chromatin condensation during spermiogenesis.


1989 ◽  
Vol 2 (2) ◽  
pp. 107-112 ◽  
Author(s):  
R. K. Iles ◽  
T. Chard

ABSTRACT Material with the immunochemical characteristics of human chorionic gonadotrophin (hCG) is produced by bladder tumour cells in vitro and in vivo. In order to characterize this material further, media were collected from 17 cell cultures (three choriocarcinomas, seven bladder carcinomas and seven 'normal' urothelium). The hCG-like material was compared with pregnancy hCG and purified α- and β-subunits by specific radioimmunoassays. Media were also submitted to affinity chromatography and the fractions further analysed by SDS-PAGE and Western blotting. It was shown that both the neoplastic and normal urothelium produced only free β-subunit-like material. This urothelial 'β-hCG' has the same molecular weight and electrophoretic mobility as that present in the intact hCG of pregnancy.


2007 ◽  
Vol 19 (1) ◽  
pp. 288
Author(s):  
M.-K. Kim ◽  
H.-J. Oh ◽  
Y. H. Fibrianto ◽  
G. Jang ◽  
H.-J. Kim ◽  
...  

Growth, maturation, and ovulation of the Graafian follicle depend on appropriate patterns of secretion, sufficient concentrations, and adequate ratios of various reproductive hormones, especially follicle stimulating hormone (FSH) and luteinizing hormone (LH) (Van Tol et al. 1996 Mol. Reprod. Dev. 45, 218–224). The present study investigated the effects of FSH and human chorionic gonadotrophin (hCG) on the nuclear maturation of canine oocytes. In addition, in order to investigate the effect of stage of the estrous cycle on the meiotic competence of canine oocytes matured in vitro, oocytes were collected from various reproductive states and matured in vitro in the presence of the gonadotrophins. Estrous cycle stage was evaluated for each bitch by ovarian morphology, and bitches were categorized according to the stage of the estrous cycle (anestrus, follicular, or diestrus) prior to oocyte collection. Recovered oocytes were cultured in serum-free tissue culture medium (TCM)-199 supplemented with various concentrations of FSH (Exp. 1: 0, 0.1, 1.0, or 10 IU) or hCG (Exp. 2: 0, 0.5, 1.0, or 10 IU) or both (Exp. 3: 1 IU FSH + 1 IU hCG) for 72 h to determine the effective concentration of these hormones, and to examine their combined effect. After maturation culture, oocytes were denuded in PBS containing 0.1% (w/v) hyaluronidase by gentle pipetting. The denuded oocytes were stained with Hoechst 33342 in glycerol and the nuclear state of oocytes was evaluated under UV light. The rates of maturation to the MII stage were significantly higher (P < 0.05) when follicular-stage oocytes were supplemented with 1 IU FSH (6.2%) compared with the other FSH-supplemented groups (0.0 to 3.3%) or to the control (1.8%), or 0.1 or 10.0 IU FSH (0 to 1.2%). Significantly higher (P < 0.05) maturation rate to MII stage was observed in follicular-stage oocytes supplemented with 1.0 IU hCG (7.2%) compared with the control or other hCG-supplemented groups (0 to 1.5%). However, FSH and hCG together did not improve the nuclear maturation rate of canine oocytes (2.4%) compared with FSH (6.2%) and hCG alone (7.2%). In conclusion, FSH or hCG alone significantly increased the maturation of canine oocytes to the MII stage. This work was supported by grant No. M1062503005-06N250300510 from KOSEF, Republic of Korea.


1980 ◽  
Vol 84 (2) ◽  
pp. 295-310 ◽  
Author(s):  
P. L. STORRING ◽  
ROSE E. GAINES-DAS ◽  
D. R. BANGHAM

The preparation and nature of the International Reference Preparation of Human Chorionic Gonadotrophin (HCG) for Immunoassay (IRP), as well as that of a second batch of ampoules (HCG 75/589) prepared identically from the same HCG preparation, are described. A collaborative study of these materials was carried out by 11 laboratories in eight countries, using different bioassay and immunoassay methods. Using the various in-vivo and in-vitro bioassays and receptor assays, the mean log potency estimates for each method within each laboratory of the HCG content of ampoules of the IRP, in terms of the Second International Standard of Human Chorionic Gonadotrophin for Bioassay (IS), were homogeneous and gave an overall weighted geometric mean (95% confidence limits) of 650 (632–669) International Units (i.u.)/ampoule. There was considerable heterogeneity of potency estimates of the IRP in terms of the IS both within and between many of the immunoassay systems (reflecting the impurity of the IS), and hence attempts to calibrate the IRP with immunoassay systems of different specificities were invalid. Immunoassay estimates of the HCG content of preparations of serum and urine, in terms of the IRP, showed considerable heterogeneity between assay systems (although the degree of this heterogeneity was no greater than that observed using the IS as standard), but the ranking order between preparations was consistent. Confirmation was obtained that contamination of the IRP with HCG-α and HCG-β subunits was insignificant. Accelerated degradation studies of the IRP stored at increased temperatures suggested that its stability under normal storage conditions would be satisfactory. It was agreed that the IRP was suitable to serve as an international reference preparation for immunoassay, and it was assigned a unitage of 650 i.u./ampoule on the basis of bioassay calibration. Since the ampoules of HCG (75/589) did not differ significantly from the IRP in any of the assay systems studied, it appeared to be equally suitable as a reference preparation. The International Reference Preparations of the α and β Subunits of Human Chorionic Gonadotrophin for Immunoassay are also described.


1963 ◽  
Vol 42 (4) ◽  
pp. 509-513 ◽  
Author(s):  
D. Gospodarowicz ◽  
J. Legault-Démare

ABSTRACT Human chorionic gonadotrophin (HCG) and lactogenic hormone (LTH or prolactin) were found practically inactive on the incorporation of 14Cacetate into cholesterol of normal rat corpus luteum in vitro. On the contrary, when added simultaneously to the incubation medium, they increased by 90% the labeling of cholesterol. When pseudopregnancy corpora lutea were used, HCG alone stimulated to the same amount, but no stimulation was observed with LTH alone. These results show that the stimulation of cholesterol synthesis is produced by a synergic action of LTH and HCG, LTH being introduced either in vivo (pseudopregnancy) or in vitro.


1985 ◽  
Vol 109 (1) ◽  
pp. 115-121
Author(s):  
Hannu Rajaniemi ◽  
Jan Sogn ◽  
Paul Holmes ◽  
Björn Källfelt ◽  
Per Olof Janson

Abstract. Pseudopregnant rats were injected with [125I] hCG, anaesthesized 1 h later and after cannulation of the aorta the ovaries were isolated and perfused with Gey & Gey buffer containing 0.2% BSA. The release of radioactivity was monitored for 2 h and analyzed by gel filtration. Five to ten per cent of the radioactivity was released within 2 h and represented small molecular weight peptides and iodotyrosine and [125I]hCG. Analysis of the ovarian radioactivity prior to and after perfusion revealed that virtually all hCG was receptor-bound. Loading the medium with unlabelled hCG displaced [125I]hCG from the receptor but did not enhance its degradation. Histological examination showed that the ovarian tissues were still intact after the 2 h perfusion. Immunohistochemical studies revealed a localization of the hCG at the cell periphery both prior to and after perfusion. These results provide evidence showing that the rate of internalization of receptor-hCG complexes in rat luteal cells is slow in vivo.


Reproduction ◽  
2005 ◽  
Vol 130 (1) ◽  
pp. 83-93 ◽  
Author(s):  
W Colin Duncan ◽  
Eva Gay ◽  
Jacqueline A Maybin

The human corpus luteum expresses genomic progesterone receptors (PRs) suggesting that progesterone may have an autocrine or paracrine role in luteal function. We hypothesised that the reduction in luteal PR reported in the late-luteal phase augmented progesterone withdrawal and had a role in luteolysis. We therefore tested the hypothesis that luteal rescue with human chorionic gonadotrophin (hCG) would maintain PR expression. PR was immunolocalised to different cell types in human corpora lutea (n = 35) from different stages of the luteal phase and after luteal rescue with exogenous hCG. There was no change in the staining intensity of theca-lutein cell or stromal cell PR throughout the luteal phase or after luteal rescue. In the late-luteal phase, granulosa-lutein cell PR immunostaining was reduced (P < 0.05) but the trend to reduction was also seen after luteal rescue with hCG (P = 0.055). To further investigate the effect of hCG on granulosa-lutein cell PR expression, an in vitro model system of cultured human luteinised granulosa cells was studied. Cells were cultured for 12–13 days exposed to different patterns of hCG and aminoglutethamide to manipulate progesterone secretion (P < 0.0001). Expression of PR A/B and PR B isoforms was examined by quantitative real-time RT-PCR. PR A/B mRNA was lower (P < 0.05) after 11–13 days of culture than after 7 days of culture. This reduction could not be prevented by hCG in the presence (P < 0.05) or absence (P < 0.05) of stimulated progesterone secretion. The expression of PR B mRNA showed a similar pattern (P = 0.054). Simulated early pregnancy in vivo and hCG treatment of luteinised granulosa cells in vitro did not appear to prevent the down-regulation of PR seen during luteolysis.


1966 ◽  
Vol 53 (3) ◽  
pp. 420-428 ◽  
Author(s):  
C. Robyn ◽  
P. O. Hubinont ◽  
E. Diczfalusy

ABSTRACT Immunologically mono-specific antisera prepared against human chorionic gonadotrophin (HCG) preparations completely neutralized in vitro as well as in vivo the luteinizing hormone (LH) and also the follicle-stimulating hormone (FSH) activity of both human hypophyseal gonadotrophin (HHG) and human menopausal gonadotrophin (HMG) preparations.


1977 ◽  
Vol 75 (1) ◽  
pp. 119-126 ◽  
Author(s):  
SOREL SULIMOVICI ◽  
M. S. ROGINSKY

The adenylate cyclase activity and the concentration of testosterone in testicular mitochondria from immature rats were measured after administration of human chorionic gonadotrophin (HCG) or dibutyryl cyclic AMP in vivo or in vitro. Intratesticular injection of HCG produced an increase in adenylate cyclase activity which preceded the rise in the level of testosterone, whereas addition of the trophic hormone in vitro resulted in simultaneous increases. Administration of dibutyryl cyclic AMP in vivo enhanced the testosterone content of the mitochondria. However, the cyclic nucleotide added in vitro at concentrations up to 5 mmol/l had no effect. Cycloheximide injected intraperitoneally before the administration of HCG abolished the stimulatory effect of the trophic hormone on the level of testosterone in the mitochondria, whereas chloramphenicol had no effect. These results, although they confirm the role of cyclic AMP as an intermediate in the stimulatory effect of HCG on the concentration of testosterone in rat testis, do not support a role for mitochondrial adenylate cyclase in this action. A protein regulator(s) formed extramitochondrially appears to be involved in the stimulatory effect of gonadotrophins on steroidogenesis.


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