AN INVESTIGATION OF THE ASSAY OF GONADOTROPHIN BY THE INDUCTION OF OVULATION IN IMMATURE MICE

SUMMARY An assay method depending on the induction of ovulation in immature mice has been applied to the assay of human urinary gonadotrophin, human chorionic gonadotrophin, pregnant mare serum gonadotrophin, and sheep and rat pituitary gonadotrophins. Valid parallel line assays were obtained in all cases, and the sensitivity and precision of the assay were such as to make it suitable for application to clinical studies. Experiments with mixtures of gonadotrophins, and the use of a barbiturate to suppress gonadotrophin secretion, indicate that the assay was not specific for LH. It is suggested that the lack of specificity may be due in part to the secretion of a variable amount of gonadotrophin by the pituitaries of the test animals.

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Effects of thymulin on spontaneous puberty and gonadotrophin-induced ovulation in prepubertal normal and hypothymic mice

Journal of Endocrinology ◽

10.1677/joe.0.1630255 ◽

1999 ◽

Vol 163 (2) ◽

pp. 255-260 ◽

Cited By ~ 12

Author(s):

L Hinojosa ◽

R Chavira ◽

R Dominguez ◽

P Rosas

Keyword(s):

Chorionic Gonadotrophin ◽

Human Chorionic Gonadotrophin ◽

Vaginal Opening ◽

Serum Progesterone ◽

Thymic Hormone ◽

Induced Ovulation ◽

Gonadotrophin Secretion ◽

Age Dependent ◽

Neuroendocrine Mechanisms ◽

Pregnant Mare Serum Gonadotrophin

The effects of thymulin administration beginning on days 19 or 24 of age on spontaneous puberty and gonadotrophin-induced ovulation were analysed in female normal and hypothymic mice. In normal and hypothymic mice, the daily administration of thymulin at 24 days of age resulted in a delay in the age of vaginal opening, with an increase in serum progesterone levels. Normal mice treated with 200 ng thymulin beginning on day 19 of age and injected with pregnant mare serum gonadotrophin (PMSG) 24 h later had an increase in ovulation rate, number of ova shed and weight of the ovaries. None of the hypothymic mice treated with thymulin on day 19 and PMSG on day 20 ovulated. PMSG treatment on day 25 induced ovulation in hypothymic mice. When these animals were injected previously with 200 ng thymulin, the number of ova shed by ovulating animals was lower than in PMSG-treated animals. Administration of thymulin and sequential injection of PMSG and human chorionic gonadotrophin 54 h later resulted in an increase in ovulatory response in comparison with those receiving only PMSG. The results suggest that thymulin plays a role in the regulation of spontaneous puberty through its effects on adrenal and ovarian endocrine functions. The increase in the ovarian PMSG response-treated animals, previously given thymulin, showed that this thymic hormone participates in the regulation of gonadotrophin secretion mechanisms and seems to be dose- and age-dependent. In hypothymic mice, neuroendocrine mechanisms regulating puberty are different from those of normal mice.

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GONADOTROPHIN REQUIREMENTS FOR IMPLANTATION IN THE MOUSE

Journal of Endocrinology ◽

10.1677/joe.0.0500019 ◽

1971 ◽

Vol 50 (1) ◽

pp. 19-27 ◽

Cited By ~ 3

Author(s):

B. M. BINDON

Keyword(s):

Luteinizing Hormone ◽

Half Life ◽

Single Injection ◽

Follicle Stimulating Hormone ◽

Chorionic Gonadotrophin ◽

Human Chorionic Gonadotrophin ◽

Suckling Mice ◽

Rat Pituitary ◽

Pituitary Glands ◽

Pregnant Mare Serum Gonadotrophin

SUMMARY Gonadotrophins were injected into mated hypophysectomized and suckling mice in an attempt to induce implantation. In these two classes of animal implantation is normally delayed by absence or suppression of pituitary gonadotrophin release. Antibodies raised against ovine gonadotrophins were injected into mice soon after mating in an attempt to inhibit implantation. Pregnant mare serum gonadotrophin (PMSG) was effective in inducing implantation in both hypophysectomized and in suckling mice. This may mean that a gonadotrophin with the qualities of PMSG normally initiates implantation. Alternatively, PMSG may have been effective by virtue of its long half-life rather than any special hormonal attributes. Human chorionic gonadotrophin was ineffective in both types of mouse. Mixtures of ovine follicle-stimulating hormone (FSH) and luteinizing hormone (LH) (100 μg of each), injected daily for 3 days, were necessary to induce implantation in hypophysectomized mice. Implantation was readily induced in suckling mice by a single injection of FSH (equivalent to 12·5 μg NIH-FSH-S3) prepared from rat pituitary glands. Implantation was readily inhibited by anti-ovine LH. Anti-ovine FSH was ineffective but this did not cross-react with mouse FSH.

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A NEW BIOASSAY METHOD FOR LUTEINIZING HORMONE DEPENDING ON THE DEPLETION OF RAT OVARIAN CHOLESTEROL

Journal of Endocrinology ◽

10.1677/joe.0.0280321 ◽

1964 ◽

Vol 28 (3) ◽

pp. 321-328 ◽

Cited By ~ 30

Author(s):

E. T. BELL ◽

S. MUKERJI ◽

J. A. LORAINE

Keyword(s):

Luteinizing Hormone ◽

High Sensitivity ◽

Chorionic Gonadotrophin ◽

Human Chorionic Gonadotrophin ◽

Assay Method ◽

Cholesterol Depletion ◽

Low Degree ◽

Extreme Sensitivity ◽

New Bioassay ◽

Pregnant Mare Serum Gonadotrophin

SUMMARY A new assay method for luteinizing hormone (LH) activity is described and the criteria for its reliability are assessed. The technique depends on ovarian cholesterol depletion in intact immature rats pretreated with pregnant mare serum gonadotrophin (PMSG) and human chorionic gonadotrophin (HCG). The main advantage of the ovarian cholesterol depletion (OCD) test is its high sensitivity, the assay being approximately five million times more sensitive than the ovarian ascorbic acid depletion (OAAD) method. The chief disadvantage of the technique is its relatively low degree of precision. The LH activity of various gonadotrophin preparations has been determined by the new OCD test. The results obtained agree well with those found by the OAAD method. The extreme sensitivity of the OCD test suggests that it will be eminently suitable for clinical use.

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INDUCTION OF OVULATION BY GONADOTROPHINS IN THE INDIAN FIVE-STRIPED PALM SQUIRREL FUNAMBULUS PENNANTI (WROUGHTON)

Journal of Endocrinology ◽

10.1677/joe.0.0390369 ◽

1967 ◽

Vol 39 (3) ◽

pp. 369-NP ◽

Cited By ~ 3

Author(s):

PUSHPA SETH ◽

M. R. N. PRASAD

Keyword(s):

Breeding Season ◽

Reproductive Cycle ◽

Single Injection ◽

Chorionic Gonadotrophin ◽

Human Chorionic Gonadotrophin ◽

Follicular Growth ◽

Induction Of Ovulation ◽

Induced Ovulation ◽

Follicular Maturation ◽

Pregnant Mare Serum Gonadotrophin

SUMMARY Superovulation was induced in palm squirrels by the administration of gonadotrophins. The regimen of treatment effective in inducing ovulation was 60 i.u. pregnant mare serum gonadotrophin (PMS) administered in three doses of 20 i.u. on days 1, 4 and 7, followed by a single injection of 40 i.u. human chorionic gonadotrophin (HCG) on day 14. The same schedule of PMS and HCG administration induced ovulation in mature and immature squirrels both during the breeding season and the period of sexual quiescence. Ovulation did not occur in PMS-treated females if HCG was not administered. The period required for follicular growth before the follicles responded to the ovulatory stimulus of HCG was 13–14 days; the interval for follicular maturation leading to the release of the ovum was approximately 24 hr. Variations in the numbers of ova shed by different groups of females are related to the phase of the reproductive cycle and the age of the squirrels.

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INHIBITION OF OVULATION IN THE GONADOTROPHIN-TREATED IMMATURE RAT BY CHLORPROMAZINE

Journal of Endocrinology ◽

10.1677/joe.0.0300087 ◽

1964 ◽

Vol 30 (1) ◽

pp. 87-95 ◽

Cited By ~ 9

Author(s):

M. X. ZARROW ◽

K. BROWN-GRANT

Keyword(s):

Wistar Rats ◽

Single Injection ◽

Chorionic Gonadotrophin ◽

Human Chorionic Gonadotrophin ◽

Assay Method ◽

Systemic Injection ◽

Immature Rat ◽

Higher Doses ◽

Effect Of Age ◽

Pregnant Mare Serum Gonadotrophin

SUMMARY The effect of age and dose of a single injection of pregnant mare serum gonadotrophin (PMS) on spontaneous ovulation in immature Wistar rats is described. Ovulation could be induced by human chorionic gonadotrophin (HCG) at least 6 days before it occurred when pregnant mare serum gonadotrophin alone was given. Chlorpromazine was shown to block pregnant mare serum gonadotrophin-induced ovulation at a dose level (0·25 mg. in a 60 g. rat) which has no effect on the ovulatory response to human chorionic gonadotrophin. Higher doses interfered with the action of injected human chorionic gonadotrophin. Ovulation could be induced in the chlorpromazine-blocked animals by the systemic injection of an extract of bovine median eminence, but the sensitivity was too low to use this response for an assay method.

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Heparin and low-dose aspirin restore placental human chorionic gonadotrophin secretion abolished by antiphospholipid antibody- containing sera

Human Reproduction ◽

10.1093/humrep/12.9.2061 ◽

1997 ◽

Vol 12 (9) ◽

pp. 2061-2065 ◽

Cited By ~ 48

Author(s):

N. Di Simone ◽

S. Ferrazzani ◽

R. Castellani ◽

S. De Carolis ◽

S. Mancuso ◽

...

Keyword(s):

Low Dose ◽

Chorionic Gonadotrophin ◽

Human Chorionic Gonadotrophin ◽

Antiphospholipid Antibody ◽

Dose Aspirin ◽

Gonadotrophin Secretion ◽

Low Dose Aspirin

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STUDIES ON A HUMAN CHORIONIC GONADOTROPHIN-LIKE MATERIAL PRESENT IN NON-PREGNANT SUBJECTS

Acta Endocrinologica ◽

10.1530/acta.0.0890492 ◽

1978 ◽

Vol 89 (3) ◽

pp. 492-505 ◽

Cited By ~ 25

Author(s):

D. M. Robertson ◽

H. Suginami ◽

H. Hernandez Montes ◽

C. P. Puri ◽

S. K. Choi ◽

...

Keyword(s):

Gel Filtration ◽

Chorionic Gonadotrophin ◽

Human Chorionic Gonadotrophin ◽

Specific Sequence ◽

Gonadotrophin Secretion ◽

Carboxy Terminal Region ◽

Assay Methods ◽

Low Levels ◽

Carboxy Terminal

ABSTRACT The presence of an hCG-like material in urinary and pituitary extracts and plasma obtained from non-pregnant subjects was investigated. Two assay methods were used to detect this material following fractionation of pituitary and urinary extracts by gel filtration (Ultrogel AcA 54) and/or isoelectrofocusing: a) a radioimmunoassay employing an antiserum raised against a specific sequence of the carboxy terminal region (residues 115– 145) of the β-subunit of hCG, and b) an in vitro bioassay method which measures both hLH and hCG activities. The fractionation procedures employed provide a satisfactory separation of highly purified hCG and hLH preparations. In the pituitary and urinary extracts hCGβ-peptide-like immunoactive (PIA) material was found consistently, which co-eluted with iodinated hCG following gel filtration and possessed pI values similar to those of hCG when subjected to isoelectrofocusing. The PIA material also exhibited in vitro biological activity similar to that shown by hLH and hCG. Detectable levels of immunoactive material were also found in plasma; however, the plasma levels of this PIA material were not influenced by classical endocrine measures such as the stimulation or inhibition of gonadotrophin secretion. The low levels of this material in plasma precluded its further characterization by gel filtration or electrofocusing. Whereas the present data and those reported by other investigators seem to suggest the presence of some hCG-like material in urinary and pituitary extracts and possibly in plasma of non-pregnant subjects, it is emphasized that the available evidence is not sufficiently conclusive to exclude other interpretations as to the nature of this material.

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Internalization of receptor-bound human chorionic gonadotrophin in preovulatory rat granulosa cells in vivo

Acta Endocrinologica ◽

10.1530/acta.0.1030406 ◽

1983 ◽

Vol 103 (3) ◽

pp. 406-412 ◽

Cited By ~ 8

Author(s):

Kalle Jääkeläinen ◽

Seppo Markkanen ◽

Hannu Rajaniemi

Keyword(s):

Plasma Membrane ◽

Granulosa Cells ◽

Subcellular Distribution ◽

Chorionic Gonadotrophin ◽

Human Chorionic Gonadotrophin ◽

Female Rats ◽

Electron Microscope Autoradiography ◽

Silver Grains ◽

Pregnant Mare Serum Gonadotrophin

Abstract. The subcellular distribution of 125I-labelled human chorionic gonadotrophin (hCG) in preovulatory rat granulosa cells was studied in vivo. Pregnant mare serum gonadotrophin-pretreated immature female rats received an iv injection of [125I]hCG a few hours before the endogenous preovulatory gonadotrophin surge. The animals were killed at 2 or 6 h after the [125I]hCG injections. Light microscope autoradiographs showed that the mural granulosa cells of large follicles were the most highly labelled cells in the ovaries. Electron microscope autoradiography was used to study the subcellular distribution of radioactivity in the mural granulosa cells. At 2 h 45% of the counted silver grains were associated with the plasma membrane and 10% with the lysosomes, at 6 h the values were 51% and 9%, respectively. The distribution of the observed silver grains was compared with the generated expected source to grain pairs by computerized linear multiple regression analysis. The magnitudes of the regression coefficients revealed that the plasma membrane and the lysosomes were the only specifically 125I-labelled organelles, that a few radioactive molecules were located diffusely over the cytoplasm at 2 h and that the 125I-radioactivity of the nuclei was negligible. The present results suggest that preovulatory rat granulosa cells are in vivo able to internalize into lysosomes [125I]hCG initially bound to LH/hCG receptors of the plasma membrane.

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