345 EFFECT OF FOLLICLE STIMULATING HORMONE AND HUMAN CHORIONIC GONADOTROPHIN ON THE IN VITRO MATURATION OF CANINE OOCYTES

2007 ◽  
Vol 19 (1) ◽  
pp. 288
Author(s):  
M.-K. Kim ◽  
H.-J. Oh ◽  
Y. H. Fibrianto ◽  
G. Jang ◽  
H.-J. Kim ◽  
...  
Keyword(s):  
Estrous Cycle ◽  
Uv Light ◽  
Follicle Stimulating Hormone ◽  
Reproductive Hormones ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Nuclear Maturation ◽  
Maturation Rate ◽  
Stimulating Hormone

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.

THE ACTION OF FOLLICLE STIMULATING HORMONE AND OF HUMAN CHORIONIC GONADOTROPHIN UPON STEROID SYNTHESIS BY RABBIT OVARIAN TISSUES IN VITRO

1964 ◽  
Vol 47 (2) ◽  
pp. 293-305 ◽  
Author(s):  
Denis Gospodarowicz
Keyword(s):  
Metabolic Pathways ◽  
Specific Activity ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Steroid Synthesis ◽  
Site Of Action ◽  
Metabolic Sequence ◽  
Stimulating Hormone

ABSTRACT Rabbit ovarian follicles have been incubated with sodium acetate-14C with and without the presence of gonadotrophin. The major radioactive metabolites found were dehydroepiandrosterone (DHEA), androst-5-ene-3β,17β-diol, testosterone, and 17α-oestradiol. Lesser amounts of radioactivity were found in pregnenolone, progesterone, androstenedione, and oestrone. The results are interpreted as evidence that two metabolic pathways exist in the follicle; the major one giving rise to androgens and oestrogens from Δ5-3β-hydroxysteroids while the other pathway involves pregnenolone and progesterone as intermediates. Follicle stimulating hormone (FSH) and human chorionic gonadotrophin (HCG) both increased the incorporation of acetate-14C into steroids and decreased the specific activity of cholesterol. The site of action of the gonadotrophins in the metabolic sequence is discussed.

Therapeutic use of gonadotrophins and clomiphene

1969 ◽  
Vol 7 (9) ◽  
pp. 33-35
Keyword(s):  
Pregnant Women ◽  
Follicle Stimulating Hormone ◽  
Therapeutic Use ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Human Pituitary ◽  
Synthetic Compound ◽  
Pituitary Glands ◽  
Post Menopausal ◽  
Stimulating Hormone

The three substances now used to stimulate the gonads in infertility are human follicle stimulating hormone (HFSH) obtained mainly from post-menopausal urine, but also from human pituitary glands, human chorionic gonadotrophin (HCG) extracted from the urine of pregnant women, and clomiphene (Clomid - Merrell), a synthetic compound which we reviewed in 1967.1

MATURATIONAL CHANGES IN SHEEP OVARIAN FOLLICLES: GONADOTROPHIC STIMULATION OF CYCLIC AMP PRODUCTION BY ISOLATED THECA AND GRANULOSA CELLS

1978 ◽  
Vol 89 (1) ◽  
pp. 166-172 ◽  
Author(s):  
T. J. Weiss ◽  
D. T. Armstrong ◽  
J. E. A. McIntosh ◽  
R. F. Seamark
Keyword(s):  
Cyclic Amp ◽  
Granulosa Cells ◽  
Follicle Stimulating Hormone ◽  
Adenosine Monophosphate ◽  
Ovarian Follicles ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Camp Production ◽  
Stimulation Of ◽  
Stimulating Hormone

ABSTRACT Theca and granulosa tissues isolated from sheep ovarian follicles of different sizes were incubated in the presence of human chorionic gonadotrophin (HCG; 5 IU/ml) or follicle stimulating hormone (FSH; 5 μg NIH-FSH-S11/ml) for 40 min. Changes in the total amounts of cyclic 3′,5′-adenosine monophosphate (cAMP) were used as an index of the responsiveness of these preparations to the hormones. Thecal tissue of both large (4–6 mm in diameter) and small (1–3 mm) follicles responded similarly to gonadotrophins. Granulosa cells from small follicles failed to respond to stimulation by HCG. FSH, however, consistently increased cAMP production in comparison with controls or cells treated with HCG. Granulosa cells of large follicles responded to both HCG and FSH.

ANTIGONADOTROPHIC PROFILE OF ANTISERA AGAINST HUMAN GONADOTROPHIN PREPARATIONS

1971 ◽  
Vol 67 (2) ◽  
pp. 262-276 ◽  
Author(s):  
P. Petrusz ◽  
C. Robyn ◽  
E. Diczfalusy
Keyword(s):  
Biological Activity ◽  
Luteinizing Hormone ◽  
Total Dose ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Immunological Activity ◽  
Human Menopausal Gonadotrophin ◽  
Stimulating Hormone

ABSTRACT Forty-two antisera were prepared in rabbits against human chorionic gonadotrophin (HCG), human hypophysial gonadotrophin (HHG), human urinary luteinizing hormone (LH) and human menopausal gonadotrophin (HMG) preparations. The gonadotrophic profiles of the antigens were previously characterized by bioassay, immunoassay and bioimmunoassay methods. The 25 most potent antisera were tested in statistically valid bioassays for their HCG and follicle stimulating hormone (FSH) neutralizing activities as well as for their neutralizing potencies against the FSH-like activity present in HCG preparations. The anti-HCG/anti-FSH ratios of the anti-HCG sera tested varied between 6.2 and > 254, while those of the anti-HHG, anti-LH and anti-HMG sera were close to 2. It was found that the total dose of immunological activity (anti-HCG neutralizing and anti-FSH neutralizing potency) rather than that of the biological activity administered to the rabbits was decisive for obtaining antisera with high anti-HCG and anti-FSH titers. Immunization with a highly purified HCG preparation (> 17 000 IU/mg) resulted in antisera exhibiting lower anti-HCG/anti-FSH ratios than did immunization with partially purified preparations. A highly purified urinary LH preparation which did not contain any detectable FSH activity gave rise to antisera exhibiting anti-HCG/anti-FSH ratios of approximately 2.0. These highly purified HCG and LH preparations were shown previously to possess high anti-FSH neutralizing potencies (Petrusz et al. 1971b). Booster injections did not change significantly the quality or the titer of the antigonadotrophic sera studied. The HCG neutralizing potency of anti-HCG sera was approximately 3 times higher when assayed against a highly purified HCG preparation (> 17 000 IU/mg) as compared to potency estimates obtained against the laboratory standard of HCG (about 2000 IU/mg). It is suggested that consideration should be given to the establishment of standard preparations of antigonadotrophic sera. It is concluded that bioimmunoassays are more suitably than conventional bioassay methods for the assessment of the antigenic purity of human gonadotrophin preparations.

Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes

Zygote ◽  
2011 ◽  
Vol 22 (1) ◽  
pp. 50-57 ◽  
Author(s):  
F. Marco-Jiménez ◽  
J.S. Vicente ◽  
M.P. Viudes-de-Castro
Keyword(s):  
Oxygen Concentration ◽  
Cell Function ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Low Oxygen ◽  
Oocyte Growth ◽  
Important Indicator ◽  
Nuclear Maturation ◽  
Defined Culture

SummaryThe choice of medium and supplements can affect meiotic regulation and may have an impact on the regulation of mammalian oocyte growth and embryonic cell function. The aim of the present study was to assess the effects of oxygen concentration and endogenous lanosterol on the in vitro maturation (IVM) media without serum and based on recombinant human chorionic gonadotrophin in prepubertal ewe oocytes. Firstly, the effect of varying oxygen concentrations (5% and 20%) during IVM in TCM-199 supplemented (4 mg/ml bovine serum albumin (BSA), 100 μM cysteamine, 0.3 mM sodium pyruvate, 0.1 UI/ml recombinant follicle-stimulating hormone (r-FSH; Gonal-F® 75 UI, Serono, Italy), 0.1 UI/ml recombinant leuteinizing hormone (r-LH; Lhadi® 75 UI, Serono, Italy) and 1 μg/ml estradiol-17β) on subsequent nuclear maturation of oocytes examined under ultraviolet light following staining with bisbenzimide (Hoechst 33342) was investigated. Secondly, two concentrations of lanosterol (0, 10 and 50 μM) were added to the IVM medium. Nuclear maturation of oocytes was examined as previously. Lipid content in oocytes, an important indicator of cytoplasmic maturity, was also measured using Nile red fluorescent stain. The results showed that low oxygen concentration affected the nuclear maturation. Similarly, a significantly higher rate of meiosis resumption was observed with 10 μM (72.3%) of lanosterol compared with the control (51.8%) or 50 μM of lanosterol (59.4%). A significantly higher content of lipids was also observed with 10 and 50 μM of lanosterol (7.3 ± 0.2 × 106 and 7.4 ± 0.2 × 106 arbitrary units of fluorescence) compared with the control (6.7 ± 0.2 × 106 arbitrary units of fluorescence). The results indicate that 10 μM lanosterol during IVM in medium without serum and based on recombinant human chorionic gonadotrophin has a positive effect on maturation of prepubertal ewe oocytes.

OBSERVATIONS ON THE ASSAY OF HUMAN URINARY FOLLICLE-STIMULATING HORMONE BY THE AUGMENTATION TEST IN MICE

1966 ◽  
Vol 35 (2) ◽  
pp. 199-206 ◽  
Author(s):  
P. S. BROWN ◽  
M. WELLS
Keyword(s):  
No Value ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
High Potency ◽  
Human Pituitary ◽  
Ovarian Weight ◽  
C3h Mice ◽  
C57bl Mice ◽  
Stimulating Hormone

SUMMARY The follicle-stimulating hormone (FSH) content of urinary gonadotrophic extracts was assayed by its effect on the ovarian weight of immature mice when given in conjunction with 40 i.u. human chorionic gonadotrophin. About three-quarters of all routine assays gave values of λ between 0·15 and 0·30. Precision was slightly increased when the material was given in three rather than in five injections. Correction of ovarian weight for body weight was either invalid or of no value in reducing variance. Removal of between-litter variance increased precision considerably. Mice of three randomly bred colonies were all satisfactory, and inbred C57BL mice were also suitable for the assay. C3H mice were less sensitive. The efficiency of different methods of extracting FSH from urine was examined. The method of Johnsen (1958) using precipitation with tannic acid was considered the most satisfactory and gave extracts of high potency and low bulk. Limited experiments in which purified human pituitary FSH was assayed with and without added luteinizing hormone, gave results compatible with the assumption that the method is specific for FSH.

PRODUCTION OF ANTISERA AGAINST HIGHLY PURIFIED HUMAN FOLLICLE-STIMULATING HORMONE, LUTEINIZING HORMONE AND THYROID-STIMULATING HORMONE

1976 ◽  
Vol 70 (3) ◽  
pp. 335-344 ◽  
Author(s):  
J. I. THORELL ◽  
B. HOLMSTRÖM
Keyword(s):  
Luteinizing Hormone ◽  
Follicle Stimulating Hormone ◽  
Thyroid Stimulating Hormone ◽  
High Titre ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Antigenic Determinants ◽  
High Avidity ◽  
Cross Reactions ◽  
Stimulating Hormone

SUMMARY Antisera were produced in rabbits against highly purified preparations of human LH (2000 or 10000 i.u./mg), human FSH (5500 i.u./mg), and human TSH (7·5 i.u./mg). Most rabbits produced antisera of high titre and high avidity. Cross-reactions were minimal between human TSH and human chorionic gonadotrophin (HCG) and between human FSH and HCG but marked between human LH and HCG. TSH and FSH also showed a constant but relatively weak cross-reaction. LH cross-reacted with FSH to a higher degree than did HCG. The avidity of the antisera was high. It was concluded that much of the lack of specificity recorded for glycoprotein antisera are effects of impure immunogens. Some of the true cross-reactions are probably explained by shared antigenic determinants of the β-subunits. Unadsorbed antisera could be used for assay of FSH and TSH in plasma from pregnant women.

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