Expression of 17β-hydroxysteroid dehydrogenase in the rat ovary during follicular development and luteinization induced with pregnant mare serum gonadotrophin and human chorionic gonadotrophin

1994 ◽  
Vol 140 (3) ◽  
pp. 409-417 ◽  
Author(s):  
S A Ghersevich ◽  
M H Poutanen ◽  
H J Rajaniemi ◽  
R K Vihko
Keyword(s):  
Granulosa Cells ◽  
Follicular Development ◽  
Hydroxysteroid Dehydrogenase ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Northern Hybridization ◽  
Enzyme Expression ◽  
Immature Rat ◽  
Rat Ovary ◽  
Pregnant Mare Serum Gonadotrophin

Abstract Antibodies against human placental 17β-hydroxysteroid dehydrogenase (17-HSD) and 17-HSD cDNA were used to study the expression of the corresponding enzyme in the immature rat ovary during follicular development and luteinization, which were induced by treating the animals with pregnant mare serum gonadotrophin (PMSG) or with PMSG followed by human chorionic gonadotrophin (hCG). Immuno-blot analysis indicated that the Mr of the 17-HSD expressed in rat granulosa cells was 35 000, as previously shown for the human placental enzyme. In immunohistochemical studies of untreated immature rat ovaries, only the granulosa cells from small antral follicles were stained. One day after PMSG treatment, strong expression of 17-HSD was observed in the granulosa cells of growing Graafian follicles. A marked decrease in enzyme expression was observed in preovulatory follicles on day 2 of PMSG treatment, starting from the basal layers of granulosa cells and progressing toward the luminal cells. No 17-HSD expression was detected in luteinized follicles or corpora lutea 22 h after hCG injection. The stroma and theca cells were negative for 17-HSD staining. In Northern hybridization analyses, two 17-HSD mRNAs were detected (1·4 and 1·7 kb). The strongest expression for both mRNAs was detected after 1 day of PMSG treatment, coinciding with maximal immunostaining of the enzyme protein. Down-regulation of 17-HSD observed by immunohistochemistry was reflected in a similar decrease in mRNA expression and the signals were almost undetectable 22 h after hCG injection. Our data suggest that 17-HSD expression in rat granulosa cells is up-regulated during follicular development and, thereafter, the enzyme expression is down-regulated during luteinization. Journal of Endocrinology (1994) 140, 409–417

CHANGES IN STEROID CONCENTRATION IN THE OVARIES OF IMMATURE RATS TREATED WITH PREGNANT MARE SERUM GONADOTROPHIN AND HUMAN CHORIONIC GONADOTROPHIN

1977 ◽  
Vol 75 (1) ◽  
pp. 43-48 ◽  
Author(s):  
S. BAUMINGER ◽  
B. ECKSTEIN ◽  
H. R. LINDNER
Keyword(s):  
Dehydrogenase Activity ◽  
Hydroxysteroid Dehydrogenase ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Immature Rat ◽  
Immature Rats ◽  
Rat Ovary ◽  
Pregnant Mare Serum Gonadotrophin

The concentrations of testosterone, progesterone and 20α-hydroxypregn-4-en-3-one (20α-OHP) were measured in the ovaries of immature rats in which ovulation was induced by treatment with pregnant mare serum gonadotrophin (PMSG) and, 48 h later, with human chorionic gonadotrophin (HCG). The concentration of testosterone in the tissue increased significantly 48 h after treatment with PMSG, reached a peak 4 h after the administration of HCG and declined to the basal level 4 h later. Increases in the levels of progesterone and 20α-OHP were observed 4 h after the administration of HCG. Whereas the level of 20α-OHP continued to rise during the subsequent 30 h, progesterone levels declined near the presumed time of ovulation (12 h after administration of HCG). It is concluded that 20α-hydroxysteroid dehydrogenase activity is present in the immature rat ovary before ovulation and that an increase in the production of testosterone in the ovaries of rats treated with PMSG and HCG precedes increased production of progesterone and 20α-OHP in these ovaries.

CXADR-like membrane protein (CLMP) in the rat ovary: stimulation by human chorionic gonadotrophin during the periovulatory period

2016 ◽  
Vol 28 (6) ◽  
pp. 742
Author(s):  
Feixue Li ◽  
Xiaoping Miao ◽  
Yonglong Chen ◽  
Thomas E. Curry
Keyword(s):  
Cell Adhesion ◽  
Protein Kinase ◽  
Membrane Protein ◽  
Mrna Expression ◽  
Granulosa Cells ◽  
Receptor Activation ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Rat Ovary ◽  
Stimulation Of

CXADR-like membrane protein (CLMP) is a novel cell–cell adhesion molecule. The present study investigated the spatiotemporal expression pattern of CLMP and its regulation in the rat ovary during the periovulatory period. Real-time polymerase chain reaction analysis revealed that Clmp mRNA was rapidly stimulated in intact ovaries by 4 h after human chorionic gonadotrophin (hCG) treatment. In situ hybridisation analysis demonstrated that Clmp mRNA expression was stimulated in theca cells at 4 h after hCG and remained elevated until 12 h. Clmp mRNA was also upregulated in granulosa cells and was present in forming corpora lutea. Our data indicate that the protein kinase A but not the protein kinase C pathway regulates the expression of Clmp mRNA in granulosa cells. Phosphatidylinositol 3 kinase and p38 kinase are also involved in regulating Clmp mRNA expression. The stimulation of Clmp mRNA by hCG requires new protein synthesis. Furthermore, inhibition of epidermal growth factor receptor activation significantly inhibited Clmp mRNA expression, whereas inhibition of prostaglandin synthesis or progesterone action had no effect. The stimulation of CLMP in the rat ovary may be important in cell adhesion events during ovulation and luteal formation such as maintaining the structure and communication of ovarian follicular and luteal cells.

Distribution of 125I-labelled follicle-stimulating hormone and human chorionic gonadotrophin in the gonads of hypogonadal (hpg) mice

1982 ◽  
Vol 93 (2) ◽  
pp. 247-NP ◽  
Author(s):  
H. M. Charlton ◽  
Dilys Parry ◽  
D. M. G. Halpin ◽  
R. Webb
Keyword(s):  
Granulosa Cells ◽  
Follicular Development ◽  
Interstitial Cells ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Testicular Development ◽  
Similar Response ◽  
Thecal Cells ◽  
Lh Rh ◽  
Lh Releasing Hormone

Hypogonadal mice are deficient in LH releasing hormone (LH-RH), the releasing factor for LH and FSH, with a consequent failure of postnatal ovarian and testicular development. After intravenous injection of hypogonadal females with 125I-labelled human chorionic gonadotrophin (hCG), followed by autoradiography of semi-thin (1 μm) slices of the ovary, labelled hCG was found to be associated with interstitial cells and thecal cells with little or no labelling of granulosa cells. Labelled human FSH was associated solely with granulosa cells. Hypogonadal females, implanted for 5 days with a silicone elastomer capsule of oestrogen, showed a similar response to that of normal females with hCG labelling of the granulosa cells of the larger follicles as well as of the thecal cell layer. Furthermore, subcutaneous injection of hypogonadal females with LH-RH (50 ng), 12 times daily for 5 days, increased uterine weight and stimulated ovarian development with some large follicles binding hCG to both thecal and granulosa cells. Therefore stimulation of follicular development may possibly be associated with increased oestradiol concentrations. In the male, after injection of 125I-labelled hCG, silver grains were associated with the interstitial cells alone in both hypogonadal and normal mice. Labelled human FSH was undetectable in semi-thin testicular sections, but the mode of injection (intravenous) may not have allowed enough labelled hormone to reach the testis in order to resolve the question as to whether the hypogonadal or normal testis can bind FSH.

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

1983 ◽  
Vol 103 (3) ◽  
pp. 406-412 ◽  
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.

THE EFFECT OF CHLORMADINONE ON THE RESPONSE OF THE OVARIES AND UTERUS OF THE IMMATURE RAT TO GONADOTROPHIC STIMULATION

1964 ◽  
Vol 30 (2) ◽  
pp. 235-245 ◽  
Author(s):  
M. J. K. HARPER
Keyword(s):  
Ovarian Response ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Female Rats ◽  
Corpora Lutea ◽  
Immature Female ◽  
Immature Rat ◽  
Orally Active ◽  
Pregnant Mare Serum Gonadotrophin ◽  
Intact Rats

SUMMARY The effects of chlormadinone (6-chloro-Δ6-17α-acetoxyprogesterone), an orally active progestational agent without significant oestrogenic activity, on the response of the ovaries of intact and hypophysectomized immature female rats to exogenous gonadotrophin have been examined. Administration of the steroid whether starting on the same day as, or 4 days before treatment with gonadotrophin, did not depress the ovarian response in intact rats. In hypophysectomized animals, pretreated with the progestagen, the ovarian response to gonadotrophin was depressed. In intact rats, treatment with the steroid and pregnant mare serum gonadotrophin (PMSG) resulted in ovulation, whereas in similar animals given PMSG alone no corpora lutea were found. Corpora lutea were seen in all groups given PMSG and human chorionic gonadotrophin (HCG) but ovulation occurred earlier when, in addition, treatment with the steroid was included. In only one experiment with intact rats did administration of the steroid alone cause a significant increase in uterine weight compared with controls. In neither experiment on hypophysectomized animals did such an increase occur, and significant decreases were recorded.

scholarly journals Early postnatal methoxychlor exposure inhibits folliculogenesis and stimulates anti-Mullerian hormone production in the rat ovary

2006 ◽  
Vol 191 (3) ◽  
pp. 549-558 ◽  
Author(s):  
Mehmet Uzumcu ◽  
Peter E Kuhn ◽  
Jason E Marano ◽  
AnnMarie E Armenti ◽  
Lisa Passantino
Keyword(s):  
Western Blot ◽  
Granulosa Cells ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Endocrine Disruptor ◽  
Follicular Development ◽  
Immature Rat ◽  
Antral Follicles ◽  
Ovarian Morphology ◽  
Rat Ovary

Methoxychlor [1,1,1-trichloro-2,2-bis(4-methoxyphenyl) ethane; MXC] is a chlorinated hydrocarbon pesticide commonly used in the United States as a replacement for DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane]. While MXC is a weak estrogenic compound, its more active, major metabolite [2,2-bis(p-hydroxyphenyl)-1,1,1-trichloroethane; HPTE] shows estrogenic, anti-estrogenic, or anti-androgenic properties depending on the receptor subtype with which it interacts. Anti-Mullerian hormone (AMH) is a paracrine factor that suppresses initial follicle recruitment in the ovary. Studies have shown the effects of exposure to MXC on adult ovarian morphology and function. However, the effect of exposure to MXC at an early postnatal stage on pre-pubertal follicular development and ovarian AMH production has not been studied. Around postnatal day (P) 4, most of the primordial follicular assembly in rats is complete, and a large number of primordial follicles transition into the primary follicle stage, a process that is inhibited by estrogen. The objective of this study was to examine the effect of early postnatal (P3–P10) MXC exposure on ovarian morphology and size, follicle number, and AMH production in the pre-pubertal (P20) rat ovary and to investigate the effect of HPTE on AMH production in immature rat granulosa cells in vitro. Female rats were injected (s.c.) daily with vehicle (control) or 1, 10, 50, 100, or 500 mg MXC/kg per day (referred to here as 1MXC, 10MXC, and so forth.) between P3 and P10. On P20, uterine and ovarian weights were determined, ovarian histology was examined, and follicles were counted and classified into primordial, primary, secondary, pre-antral, or antral stages using the two largest serial sections at the center of the ovary. Ovarian AMH production was examined using immunohistochemistry and western blot analysis. The effect of HPTE (0.5–25 μM) on AMH production in cultured immature rat granulosa cells was determined by western blot analysis. Ovarian weight was reduced by 50, 100, and 500MXC (P < 0.01). MXC treatment inhibited folliculogenesis. Both 100 and 500MXC had a reduced number of antral follicles (P < 0.05) with a concomitant increase in pre-antral follicles (P < 0.05). Follicle numbers were not significantly affected by 1, 10, or 50MXC. Total follicle number and the number of primordial, primary, or secondary stage follicles were not significantly different in all treatment groups. Immunohistochemistry showed that MXC-treated ovaries had more AMH-positive follicles with stronger AMH immunostaining. Western blot analysis showed that AMH production was 1.6 ± 0.2, 1.85 ± 0.6, and 2.2 ± 0.5 times higher in the 50, 100, and 500MXC ovaries as compared with the control ovaries respectively (P < 0.05). Granulosa cells treated with 1 or 5 μM HPTE had significantly greater AMH production (P < 0.05). These results demonstrate that MXC inhibits early ovarian development and stimulates AMH production directly in the rat ovary. In addition, HPTE was shown to stimulate AMH production in rat granulosa cells. Endocrine disruptors are widespread in the environment, and MXC represents a model endocrine disruptor due to the multiple actions of its metabolites. This study confirms that the endocrine disruptor MXC inhibits follicular development and demonstrates for the first time that MXC and HPTE directly stimulate AMH productionin the ovary. Thisnovel finding suggests that elevated AMH may play a role in MXC’s inhibitory effect in the ovary.

OVULATION INDUCED BY PREGNANT MARE SERUM GONADOTROPHIN IN THE IMMATURE RAT TREATED NEONATALLY WITH A LOW OR A HIGH DOSE OF ANDROGEN

1972 ◽  
Vol 55 (3) ◽  
pp. 533-541 ◽  
Author(s):  
J. Th. J. UILENBROEK ◽  
J. J. van der WERFF ten BOSCH
Keyword(s):  
Testosterone Propionate ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Female Rats ◽  
High Dose ◽  
Immature Female ◽  
Immature Rat ◽  
Progesterone Treatment ◽  
Combined Administration ◽  
Pregnant Mare Serum Gonadotrophin

SUMMARY Ovulation-inducing effects of pregnant mare serum gonadotrophin (PMSG) were studied in immature female rats treated on day 5 (day 1 = day of birth) with oil or with 5 or 1250 μg testosterone propionate (TP). The response of rats treated with 1250 μg TP was negligible regardless of the age of the animals and of the dose of PMSG. The response of rats treated with 5 μg TP to PMSG alone was low (36% of rats, with 2·6 ova/ovulating rat), but could be improved by progesterone administration 2 days after PMSG injection (91% of rats, with 14·5 ova/ovulating rat). At every age and dose of PMSG tested the response of animals treated with 5 μg TP to combined PMSG and progesterone treatment was less than that of control animals. It is concluded that neonatal TP treatment diminishes the release of endogenous ovulating hormone subsequent to PMSG injection. This effect is dependent on the dose of TP used, but already demonstrable in animals treated with 5 μg TP on day 5, which would have been cyclic and fertile after puberty. Only for the animals treated with 1250 μg TP could a decreased sensitivity of the ovaries to combined administration of PMSG and human chorionic gonadotrophin be demonstrated.

STUDIES ON GONADOTROPHIN-INDUCED OVULATION IN THE IMMATURE RAT

1968 ◽  
Vol 41 (2) ◽  
pp. 171-177 ◽  
Author(s):  
E. T. BELL ◽  
S. F. LUNN
Keyword(s):  
Marked Effect ◽  
Cell Mass ◽  
Hormone Treatment ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Maximum Increase ◽  
Immature Rat ◽  
Immature Rats ◽  
Wet Weight ◽  
Pregnant Mare Serum Gonadotrophin

SUMMARY The effect of the administration of 25 i.u. human chorionic gonadotrophin (HCG) on the induction of ovulation in intact immature rats treated with 50 i.u. pregnant mare serum gonadotrophin (PMSG) has been studied. After the administration of HCG a marked increase in ovarian wet weight was observed. The maximum increase, which occurred 10 hr. after hormone treatment, was noted 2 hr. before ova were found in the oviducts. The alteration in ovarian wet weight was associated with a fall in percentage solids. However, it appears likely that an increase both in follicular fluid and in cell mass occurred before ovulation. Possible reasons for the absence of any marked effect on uterine wet weight or percentage solids are discussed.

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

1964 ◽  
Vol 30 (1) ◽  
pp. 87-95 ◽  
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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