The Fine Structure of Rat Granulosa Cell Cultures Correlated with Progestin Secretion

1973 ◽  
Vol 31 ◽  
pp. 552-553
Author(s):  
T. M. Crisp ◽  
F.R. Denys
Keyword(s):  
Fine Structure ◽  
Granulosa Cell ◽  
Cell Cultures ◽  
Single Injection ◽  
Surrounding Medium ◽  
Petri Dish ◽  
Female Rats ◽  
Vaginal Smear ◽  
Immature Female ◽  
Serum Gonadotropin

The purpose of this paper is to present observations on the fine structure of rat granulosa cell cultures grown in the presence of an adenohypophyseal explant and to correlate the morphology of these cells with progestin secretion. Twenty-six day old immature female rats were given a single injection of 5 IU pregnant mares serum gonadotropin (PMS) in order to obtain ovaries with large vesicular follicles. At 66 hrs. post-PMS administration (estrus indicated by vaginal smear cytology), the ovaries were removed and placed in a petri dish containing medium 199 and 100 U penicillin/streptomycin (P/S)/ml. Under a 20X magnification dissecting microscope, some 5-8 vesicular follicles/ovary were punctured and the granulosa cells were expressed into the surrounding medium. The cells were transferred to centrifuge tubes and spun down at 1000 rpm for 5 mins.

Expression and regulation of high mobility group AT-hook 1 (HMGA1) during ovulation and luteinisation in rat ovary

2019 ◽  
Vol 31 (4) ◽  
pp. 698 ◽  
Author(s):  
Hao-ran Li ◽  
Yan Li ◽  
Yu Liu ◽  
Jiao-jiao Yu ◽  
Fei-xue Li
Keyword(s):  
Mrna Expression ◽  
Granulosa Cell ◽  
Cell Cultures ◽  
Granulosa Cells ◽  
In Situ Hybridisation ◽  
High Mobility ◽  
High Mobility Group ◽  
Female Rats ◽  
Corpora Lutea ◽  
Hmga1 Expression

High mobility group AT-hook 1 (HMGA1) is able to regulate gene expression and function as a tumour suppressor. The spatiotemporal expression pattern of HMGA1 was investigated in this study. Immature female rats (22–23 days old) were treated with 10IU, s.c., pregnant mare’s serum gonadotrophin to stimulate follicular development, followed 48h later by injection with 5IU, s.c., human chorionic gonadotrophin (hCG). Whole ovaries or granulosa cells were collected at various times after hCG administration (n=3 per time point). Real-time polymerase chain reaction and western blot analysis revealed that HMGA1 was highly stimulated in the ovary by 4–12h after hCG treatment. In situ hybridisation analysis demonstrated that Hmga1 mRNA expression was induced in granulosa cells between 8 and 12h after hCG treatment. There was negligible Hmga1 mRNA signal observed in newly forming corpora lutea. In addition, the data indicated that both the protein kinase (PK) A and PKC pathways regulated Hmga1 expression in rat granulosa cells. In rat granulosa cell cultures, upregulation of Hmga1 was dependent on new protein synthesis because Hmga1 was inhibited by cycloheximide. Furthermore, Hmga1 mRNA expression in rat granulosa cell cultures was inhibited by AG1478, whereas NS398 and RU486 had no effect, suggesting that Hmga1 expression was regulated, in part, by the epidermal growth factor pathway. In summary, the findings of this study suggest that induction of Hmga1 may be important for theca and granulosa cell differentiation into luteal cells.

Eremomastax speciosa potentializes the PMSG-inducing effect on some physiological and biochemical parameters in PMSG-primed immature rats

Zygote ◽  
2020 ◽  
Vol 28 (6) ◽  
pp. 482-488
Author(s):  
Gildas Tetaping Mbemya ◽  
Marie Stéphanie Goka Chekem ◽  
Landry Lienou Lienou ◽  
Njina Nguedia Sylvain ◽  
Jiatsa Nathalie Donfack ◽  
...  
Keyword(s):  
Biochemical Parameters ◽  
Methylene Chloride ◽  
Relative Weight ◽  
Female Rats ◽  
Immature Female ◽  
Pregnant Mare Serum Gonadotropin ◽  
Serum Gonadotropin ◽  
Puberty Onset ◽  
Uterine Proteins ◽  
Physiological And Biochemical

SummaryThe present study evaluated the effect of the aqueous extract from leaves of E. speciosa on some physiological and biochemical parameters of reproduction and the onset of puberty in pregnant mare serum gonadotropin (PMSG)-primed immature female rats. High pressure liquid chromatography (HPLC) was used to quantify the phenolic compounds in the methanol/methylene chloride (1:1) extract, the ethanolic and ethyl acetate fractions and the aqueous residue of E. speciosa. E. speciosa (0, 8, 32 or 64 mg/kg) were administered for 15 days to 24 non-PMSG-primed and 24 primed rats with 0.01 IU of PMSG. At the end of the treatment period, animal were sacrificed and their body, ovarian, uterine weight, ovarian protein or cholesterol level, as well as data on puberty onset were recorded. Of the 16 polyphenolic compounds quantitatively revealed in the extracts and fractions of E. speciosa after HPLC analysis, quercetin, rutin, apigenin and eugenol were the most abundant. Non-primed rats showed a significant increase (P < 0.05) in the uterine relative weight at the dose of 8 mg/kg when compared with the other treatments. The uterine proteins and the ovarian cholesterol (P < 0.05), respectively, showed a reduction at doses of 64 mg/kg and 32 mg/kg in non-primed rats. However in PMSG-primed rats, a significant decrease (P < 0.05) was observed in ovarian cholesterol at 64 mg/kg. In conclusion, E. speciosa potentializes the PMSG-inducing effect on folliculogenesis in PMSG-primed rats.

Ontogeny of sex differences in LH and FSH levels 48 h after castration in the rat

1981 ◽  
Vol 241 (6) ◽  
pp. E460-E464
Author(s):  
J. R. Lorenzen ◽  
J. A. Ramaley
Keyword(s):  
Follicle Stimulating Hormone ◽  
Female Rats ◽  
Hormonal Changes ◽  
Vaginal Opening ◽  
Immature Female ◽  
Immature Male ◽  
Male And Female Rats ◽  
Serum Lh ◽  
Pituitary Secretion ◽  
Serum Gonadotropin

Serum gonadotropin levels were measured 12, 24, and 48 h after gonadectomy in male and female rats (ages, 22--60 days) to assess when during development the rate of rise of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) after castration approximates that seen in the gonadectomized adult. In females serum LH levels 48 h after ovariectomy were increased above sham levels only when the ovaries were removed prior to vaginal opening. Ovariectomy on the day of vaginal opening or at older ages resulted in no increase in LH levels by 48 h after surgery. Serum FSH levels at 24 and 48 h after ovariectomy declined with increasing age at the time of ovariectomy. In males serum LH levels at 48 h after castration increased with increasing age at the time of gonadectomy. Serum FSH levels at either 12, 24, or 48 h after orchidectomy did not change appreciably with age at the time of surgery. It is concluded that the acute pituitary secretion of gonadotropins after removal of testes in the immature male resembles that seen in the mature male early in the course of the development of sexual maturity. In contrast, the acute pituitary secretion of gonadotropins after removal of the ovaries in the immature female does not resemble that seen in the ovariectomized adult until she is mature and capable of ovulating. Thus, the observed delay in the rise of LH seen in ovariectomized adults may be a function of some aspect of the hormonal changes associated with the estrous cycle.

Induction of ovulation in immature female rats by a single injection of 4-hydroxyoestradiol-dibenzoate

1982 ◽  
Vol 100 (1) ◽  
pp. 105-108 ◽  
Author(s):  
Günter Emons ◽  
Peter Ball
Keyword(s):  
Single Injection ◽  
Female Rats ◽  
High Dose ◽  
Corpora Lutea ◽  
Immature Female ◽  
Induction Of Ovulation ◽  
Oestradiol Benzoate ◽  
Different Doses

Abstract. This study was designed to test the ability of 2- and 4-hydroxyoestrogens to induce ovulations and the formation of corpora lutea in immature female rats. To this end 25 day old animals received a single injection of different doses of either 4-hydroxyoestradiol-dibenzoate (10, 25, 50 μg) or 2-hydroxyoestradiol-dibenzoate (25, 250 μg) or oestradiol-benzoate (10, 25, 50 μg). On day 31 the ovaries were checked for corpora lutea. 4-Hydroxyoestradiol-dibenzoate and oestradiol-benzoate in doses of 50 or 25 μg significantly increased the number of animals with corpora lutea whereas animals treated with 10 μg of either steroid did not differ significantly from the respective vehicle groups. 2-Hydroxyoestradioldibenzoate, even at the high dose of 250 μg did not show a significant effect. This is the first demonstration that a catecholoestrogen 4-hydroxyoestradiol - can induce ovulation. As its potency in doing so is similar to that of oestradiol and as 4-hydroxyoestradiol can be formed in neuroendocrine tissues from primary oestrogens, we conclude that this catecholoestrogen might play a role in the regulation of ovulation.

INCREASED PRODUCTION OF FOLLICLE-STIMULATING HORMONE IN IMMATURE FEMALE RATS BY A SINGLE INJECTION OF ANDROGEN OR OESTROGEN

1969 ◽  
Vol 45 (1) ◽  
pp. 29-36 ◽  
Author(s):  
R. H. NAQVI ◽  
D. C. JOHNSON
Keyword(s):  
Testosterone Propionate ◽  
Single Injection ◽  
Follicle Stimulating Hormone ◽  
Ovarian Response ◽  
Steroid Treatment ◽  
Female Rats ◽  
Immature Female ◽  
Ovarian Weight ◽  
Oestradiol Benzoate ◽  
Stimulating Hormone

SUMMARY An increase in ovarian weight in immature rats after the injection of chorionic gonadotrophin (HCG) was used to measure variations in endogenous follicle-stimulating hormone (FSH) after steroid treatment. A single injection of several steroids (testosterone, androstenediol, androstenedione, oestradiol benzoate) given 12–96 hr. before treatment with HCG caused a 30–200% increase in ovarian weight. This was not a direct effect of the steroids since hypophysectomy abolished the response, and administration of the compounds concurrently with HCG was ineffective. Within certain limits an increase in the duration of pretreatment enhanced the ovarian response while an increase in the dose of steroid had little effect. Pretreatment with testosterone propionate did not change pituitary FSH activity, indicating that the increase in circulating FSH was due to an increased production of hormone. On the other hand, pituitary FSH in animals treated with oestradiol benzoate was significantly lowered within 72 hr. suggesting an increased release of FSH.

scholarly journals Regulation of Ascorbic acid by gonadotropins in rat Ovary

2019 ◽  
Author(s):  
Shah Saddad Hussain ◽  
Taruna Arora ◽  
Madan Mohan Chaturvedi ◽  
Sharmila Basu-Modak ◽  
Rajesh Chaudhary ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Specific Activity ◽  
Inhibitory Effect ◽  
Female Rats ◽  
Transcriptional Level ◽  
Immature Female ◽  
Serum Gonadotropin ◽  
Rat Ovary ◽  
Dose Dependent

AbstractThe dose dependent depletion of ovarian Ascorbic acid (AA) in rat ovaries, has been used as a bioassay for measurement of Luteinizing Hormone (LH). However, the mechanism of action of gonadotropin (LH, FSH) on ascorbic acid depletion is not completely clear in biochemical terms. To elucidate the mechanism, we looked for the pathways; one, where L-GulonateDehydrogenase (L-GuDH) catalyzes the conversion of L-Gulonic acid (L-GuA) to L-Xylulose, and, in the second the pathway conversion of L-GuA to AA, in a cats, dogs and Rats. Kinetic analysis of the enzyme L-GuDHin vitroshowed the inhibitory effect of AA on L-GuDH. Therefore, we hypothesized that gonadotropins (FSH and LH) may regulate the L-GuDH maintain level of AA in ovary. LH administration to super-ovulated immature female rats caused depletion of ovarian AA but did not result in any change in the specific activity of the ovarian L-GuDH. Further, we administrated a surrogate FSH like hormone (PMSG) to immature female rats which, resulted in increased specific activity of ovarian L-GuDH. However, microarray data on RNA from ovaries exposed to FSH like hormone such as Pregnant Mare serum Gonadotropin (PMSG) did not reveal any increased expression of L-GuDH transcript. It is therefore concluded from the results obtained that; that neither LH, in decreasing the ovarian AA, nor FSH, in increasing the ovarian AA do so by regulating the activity of enzyme L-GuDH at transcriptional level. The results obtained have also been discussed by giving emphasis on the mechanism of ovarian ascorbic acid regulation of LH and FSH.

Effects of hydroxyflutamide on rats treated with a superovulatory dose of pregnant mare serum gonadotropin

1991 ◽  
Vol 69 (2) ◽  
pp. 185-190 ◽  
Author(s):  
Frank H. Yu ◽  
Young W. Yun ◽  
Basil Ho Yuen ◽  
Young S. Moon
Keyword(s):  
Serum Levels ◽  
Treated Group ◽  
Biologically Active ◽  
Female Rats ◽  
Vehicle Group ◽  
Immature Female ◽  
Pregnant Mare Serum Gonadotropin ◽  
Serum Gonadotropin ◽  
17Β Estradiol ◽  
Androgen Binding

Immature female rats treated with superovulatory doses of pregnant mare serum gonadotropin (PMSG) were used to study the effects of the antiandrogen hydroxyflutamide on steroid production, particularly the biologically active androgens, in two experiments. In the first experiment, animals were given either 5 mg hydroxyflutamide or vehicle alone at 30 and 36 h following 40 IU PMSG. Compared with the vehicle group, hydroxyflutamide treatment significantly reduced the percentage of degenerate oocytes recovered from oviducts (p < 0.05). Serum levels of testosterone and androstenedione, and their aromatized product 17β-estradiol, significantly decreased (p < 0.05) in the hydroxyflutamide-treated group; however, nonaromatizable androgen, 5α-dihydrotestosterone, was not affected. In the second experiment, ovaries obtained 48 h after stimulation with 4 or 40 IU PMSG were incubated with and without hydroxyflutamide (10−5 M) and (or) testosterone (10−7 M) to study [4-14C]pregnenolone metabolism to major steroids. In 40 IU stimulated ovaries, hydroxyflutamide significantly decreased the metabolism of pregnenolone to progesterone (p < 0.01) and androstenedione (p < 0.01), while the production of 17β-estradiol increased significantly (p < 0.05); however, pregnenolone conversions to testosterone and 5α-dihydrotestosterone were not affected. Testosterone completely reversed the hydroxyflutamide-induced alteration of pregnenolone metabolism. In contrast, there was no difference in the pregnenolone conversion patterns between untreated and hydroxyflutamide or hydroxyflutamide plus testosterone groups in 4 IU stimulated ovaries. Present results confirm our previous finding that hydroxyflutamide decreases the percentage of abnormal oocytes recovered from superovulating rats and indicates that this hydroxyflutamide effect may be partly mediated by altered ovarian steroidogenesis following inhibition of androgen binding in the ovary.Key words: superovulation, pregnant mare serum gonadotropin, antiandrogen, hydroxyflutamide, androgen.

Morphological Changes in the Rat Granulosa Cell Surface During Differentiation Induced by Estradiol and Gonadotropins

1977 ◽  
Vol 35 ◽  
pp. 484-485
Author(s):  
P. Bagavandoss ◽  
JoAnne S. Richards ◽  
A. Rees Midgley
Keyword(s):  
Cell Surface ◽  
Granulosa Cell ◽  
Morphological Changes ◽  
Follicular Development ◽  
Female Rats ◽  
Functional Changes ◽  
Group 4 ◽  
Group 3 ◽  
Group 5 ◽  
Group 2

During follicular development in the mammalian ovary, several functional changes occur in the granulosa cells in response to steroid hormones and gonadotropins (1,2). In particular, marked changes in the content of membrane-associated receptors for the gonadotropins have been observed (1).We report here scanning electron microscope observations of morphological changes that occur on the granulosa cell surface in response to the administration of estradiol, human follicle stimulating hormone (hFSH), and human chorionic gonadotropin (hCG).Immature female rats that were hypophysectcmized on day 24 of age were treated in the following manner. Group 1: control groups were injected once a day with 0.1 ml phosphate buffered saline (PBS) for 3 days; group 2: estradiol (1.5 mg/0.2 ml propylene glycol) once a day for 3 days; group 3: estradiol for 3 days followed by 2 days of hFSH (1 μg/0.1 ml) twice daily, group 4: same as in group 3; group 5: same as in group 3 with a final injection of hCG (5 IU/0.1 ml) on the fifth day.

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