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1.
Gen Comp Endocrinol ; 166(2): 436-42, 2010 Apr 01.
Article in English | MEDLINE | ID: mdl-20067799

ABSTRACT

To address the potential luteolytic role for prostaglandin F(2 alpha) (PGF(2 alpha)) in the corpus luteum of the common marmoset monkey (Callithrix jacchus), the ability of marmoset luteal cells, maintained in monolayer culture, to produce PGF(2 alpha) was determined in vitro in the presence and absence of human chorionic gonadotrophin (hCG) and other established pharmacological modulators of PGF(2 alpha) synthesis. We also assessed the effects of the PGF(2 alpha) analogue, cloprostenol, on progesterone output from luteal cells isolated in the early luteal phase versus the mid-luteal phase (days 3 and 14 post ovulation, respectively). Cloprostenol had no effect on progesterone output from luteal cells isolated on day 3 of the luteal phase, whereas it significantly inhibited both basal and hCG-stimulated progesterone synthesis by day 14 luteal cells during the culture period 48-72 h (P<0.001). Intra-luteal PGF(2 alpha) concentrations were 5-fold higher in luteal cells isolated in the early luteal phase than in mid-luteal phase cells (16.5+/-3.5 versus 3.5+/-0.6 pmol/10(5) cells). While PGF(2 alpha) production was unaffected by hCG in vitro, it was decreased by indomethacin (1000 ng/ml) (P<0.05) and stimulated by the calcium ionophore A23187 (10 micromol/l) (P<0.05) in luteal cells from both stages of the luteal phase. Phospholipase A(2) did not influence PGF(2 alpha) production by day 3 luteal cells whereas at 10 IU/ml, it significantly stimulated PGF(2 alpha) production by day 14 luteal cells (P<0.05). Hence, the timing of luteolysis in the common marmoset monkey appears to involve changes in both the luteal cell response to and production of PGF(2 alpha).


Subject(s)
Callithrix/physiology , Dinoprost/biosynthesis , Dinoprost/physiology , Gonadotropins/antagonists & inhibitors , Luteal Cells/physiology , Luteal Phase/physiology , Animals , Cells, Cultured , Chorionic Gonadotropin/administration & dosage , Chorionic Gonadotropin/antagonists & inhibitors , Cloprostenol/administration & dosage , Dinoprost/analysis , Female , Luteal Cells/chemistry , Luteal Cells/drug effects , Luteolysis/physiology , Luteolytic Agents/administration & dosage , Progesterone/biosynthesis , Time Factors
2.
Res Vet Sci ; 62(2): 87-91, 1997.
Article in English | MEDLINE | ID: mdl-9243703

ABSTRACT

The ability of melatonin to stimulate luteal steroidogenesis directly in the ewe was investigated by using a perfusion cannula system in vivo and luteinised granulosa cells in vitro. Exposure to human chorionic gonodotrophin (hCG) or ovine luteinising hormone (oLH), as positive controls, significantly stimulated progesterone secretion by the perfused corpus luteum within 20 minutes and by granulosa cells after two days of culture. Melatonin perfused through the corpus luteum at a physiological concentration, significantly stimulated progesterone secretion but the response was not as rapid as that to hCG. The significant stimulation of progesterone by melatonin in the perfusion system was maintained for between 30 and 60 minutes. Melatonin significantly stimulated progesterone production by granulosa cells in vitro after two days of culture, but the degree of response depended on the size of the follicles from which the cells were obtained. These results demonstrate that melatonin can act directly on the corpus luteum to increase progesterone production, and that its mechanism of action appears to be different from that of hCG or oLH in the ewe. This action of melatonin may be related to the reported improved luteal function late in the breeding season after the prolonged exposure of ewes to melatonin.


Subject(s)
Corpus Luteum/metabolism , Granulosa Cells/metabolism , Melatonin/pharmacology , Progesterone/biosynthesis , Sheep/metabolism , Animals , Cells, Cultured , Chorionic Gonadotropin/pharmacology , Corpus Luteum/cytology , Corpus Luteum/drug effects , Dose-Response Relationship, Drug , Estrus , Female , Granulosa Cells/cytology , Granulosa Cells/drug effects , Humans , In Vitro Techniques , Light , Luteinizing Hormone/pharmacology , Progesterone/metabolism , Seasons , Time Factors
3.
Biol Reprod ; 50(6): 1394-402, 1994 Jun.
Article in English | MEDLINE | ID: mdl-8080927

ABSTRACT

There is an increase in plasma concentrations of immunoreactive (ir) inhibin unaccompanied by a rise in plasma progesterone during early pregnancy in the marmoset monkey. We investigated the potential involvement of hCG and prostaglandin E2 (PGE2) in stimulating a selective increase in inhibin concentrations by measuring the production of ir-alpha-inhibin and progesterone by dispersed luteal cells cultured under serum-free conditions. After one day, hCG had no effect on progesterone production by the cells but stimulated a significant increase (p < 0.05) in alpha-inhibin production. PGE2 significantly increased progesterone production (p < 0.001) but inhibited the production of alpha-inhibin (p < 0.001). After three days of culture, output of alpha-inhibin fell to low levels and no significant effect of hCG or PGE2 was detected. Progesterone also fell with time in culture, but hCG maintained production resulting in a significant increase above control levels (p < 0.001). The addition of low density lipoproteins (LDL) to the culture medium increased progesterone production (p < 0.001) while decreasing alpha-inhibin production (p < 0.01). Immunoneutralization of endogenous alpha-inhibin resulted in a significant decrease in both basal (p < 0.05) and gonadotropin-stimulated (p < 0.05) progesterone concentrations. These results provide further evidence for differential control of progesterone and alpha-inhibin production by marmoset luteal cells and show that hCG can selectively stimulate alpha-inhibin production. In addition, alpha-inhibin may have a local paracrine action in the marmoset CL, enhancing both basal and gonadotropin-stimulated progesterone secretion.


Subject(s)
Chorionic Gonadotropin/pharmacology , Inhibins/pharmacology , Luteal Cells/metabolism , Progesterone/biosynthesis , Animals , Callithrix , Cells, Cultured , Culture Media , Dinoprostone/pharmacology , Epidermal Growth Factor/pharmacology , Female , Follicle Stimulating Hormone/pharmacology , Lipoproteins, LDL/pharmacology , Luteal Cells/drug effects , Progesterone/blood , Time Factors
5.
Am J Primatol ; 32(3): 187-195, 1994.
Article in English | MEDLINE | ID: mdl-31936919

ABSTRACT

This study describes the peripheral concentrations of immunoreactive (ir) inhibin, progesterone, and bioactive gonadotrophin during pregnancy in the marmoset monkey, a New World primate. Blood samples were taken every two weeks from six animals from ovulation to parturition. Plasma ir-inhibin concentrations were measured by inhibin α-subunit-directed radioimmunoassay (RIA) and a recently developed two-site immunoradiometric assay (IRMA) which is specific for inhibin αß dimer. Concentrations of α-inhibin increased (P < 0.001) during early pregnancy to reach a peak on week 12 of pregnancy and showed a positive correlation with bioassayable gonadotrophin concentrations (r = 0.5, n = 64; P < 0.001). The concentrations of both α-inhibin and gonadotrophin showed no further peaks and declined (P < 0.001) to low levels prior to birth. Concentrations of dimeric inhibin were substantially lower than those measured by RIA and did not vary significantly during pregnancy. Progesterone concentrations remained at luteal-phase levels during the first half of pregnancy and increased (P < 0.05) during the second half to reach maximum concentrations just prior to birth. The relationship between α-inhibin, gonadotrophin, and progesterone suggests that the increase in α-inhibin may be luteal in source and under the control of gonadotrophin. The absence of a second increase in α-inhibin later in pregnancy and the finding that lategestation placenta contained very little α-inhibin differs from observations in Old World primates studied and suggests that the placenta may not be a source of inhibin during pregnancy in the marmoset. The finding of high levels of α-inhibin, but not dimeric inhibin, suggests that inhibin-related molecules may have a role other than suppression of pituitary folliclestimulating hormone. © 1994 Wiley-Liss, Inc.

6.
Mol Cell Endocrinol ; 97(1-2): 81-91, 1993 Nov.
Article in English | MEDLINE | ID: mdl-8143907

ABSTRACT

It is well documented that prostaglandin F2 alpha (PGF2 alpha) inhibits progesterone production in luteal cells, but its mode of action is uncertain. It has recently been suggested that PGF2 alpha acts by activating the calcium and phospholipid-dependent protein kinase, protein kinase C (PKC). This hypothesis has been tested by comparing the site and mode of action of PGF2 alpha, a PGF2 alpha analogue (cloprostenol) and the PKC activator phorbol myristate acetate (4 beta PMA) in human granulosa-lutein cells. PGF2 alpha and cloprostenol exerted similar concentration-dependent inhibitory actions on gonadotrophin-stimulated cyclic AMP (cAMP) accumulation and progesterone production by human granulosa-lutein cells. The similarity in the actions of PGF2 alpha and cloprostenol in human granulosa-lutein cells suggests that they can be used interchangeably to study the role of PGF2 alpha in the regulation of steroidogenesis in the human ovary. Gonadotrophin-stimulated cAMP accumulation and progesterone production was also concentration-dependently inhibited by 4 beta PMA. In addition, cloprostenol and 4 beta PMA also inhibited dibutyryl cAMP-stimulated progesterone production, suggesting that these compounds inhibit LH action at sites before and after the generation of cAMP. The pre-cAMP site of action can be localised to the stimulatory G-protein (Gs) as both compounds inhibited cholera toxin-stimulated cAMP accumulation without affecting forskolin-stimulated cAMP accumulation. The post cAMP site of action can be localised to actions on cholesterol side chain cleavage enzyme, as both cloprostenol and 4 beta PMA inhibited 22R hydroxycholesterol-supported progesterone production without affecting pregnenolone-supported progesterone production. The finding that cloprostenol and 4 beta PMA interact with the steroidogenic cascade in a similar manner is indicative of a shared common mediator of their actions in human granulosa-lutein cells, i.e. PKC. The inhibitory actions of PGF2 alpha and 4 beta PMA on hLH-stimulated progesterone production were abolished in the presence of the PKC inhibitor, staurosporine. In addition, in PKC-depleted cells (achieved by exposure to 4 beta PMA for 20 h) the inhibitory actions of PGF2 alpha and 4 beta PMA were abolished. These results support the hypothesis that the inhibitory actions of PGF2 alpha are mediated by PKC in human granulosa-lutein cells.


Subject(s)
Dinoprost/pharmacology , Granulosa Cells/drug effects , Protein Kinase C/physiology , Bucladesine/pharmacology , Calcium/metabolism , Cells, Cultured , Cholera Toxin/pharmacology , Cloprostenol/pharmacology , Colforsin/pharmacology , Cyclic AMP/biosynthesis , Female , Humans , Hydroxycholesterols/pharmacology , Luteinizing Hormone/antagonists & inhibitors , Pregnenolone/pharmacology , Progesterone/biosynthesis , Tetradecanoylphorbol Acetate/pharmacology
7.
J Endocrinol ; 138(2): 291-8, 1993 Aug.
Article in English | MEDLINE | ID: mdl-8228738

ABSTRACT

Progesterone production by dispersed luteal cells obtained from the marmoset monkey on day 14 after ovulation can be stimulated by both prostaglandin F2 alpha (PGF2 alpha) and its structural analogue, cloprostenol. To establish whether these responses can be attributed to cross-reaction with the prostaglandin E2 (PGE2) receptor, this study compared the involvement of cyclic adenosine-3',5'-monophosphate (cAMP) and protein kinase C (PKC) in the luteotrophic responses to PGE2, PGF2 alpha and cloprostenol. While all three prostaglandins stimulated similar increases in progesterone production (239.5 +/- 7.9% of control; P < 0.01), only PGE2 stimulated a significant increase in cAMP accumulation (373.2 +/- 28.4% of control; P < 0.01). This study is the first to demonstrate PKC activity in the marmoset ovary. Following down-regulation of PKC with a tumour-promoting phorbol ester, 4 beta-phorbol 12-myristate 13-acetate (4 beta-PMA), basal progesterone production was significantly increased (150.9 +/- 8.2% of control; P < 0.05) and the luteotrophic effects of PGF2 alpha and cloprostenol were no longer evident, whereas the response to PGE2 was unaffected. These observations are consistent with the differential involvement of cAMP and PKC in the luteotrophic responses to PGE2 and PGF2 alpha/cloprostenol respectively. Hence, we conclude that the luteotrophic actions of prostaglandins E2 and F2 alpha on dispersed marmoset luteal cells are mediated via different receptors and signal transduction pathways.


Subject(s)
Corpus Luteum/drug effects , Cyclic AMP/metabolism , Dinoprost/pharmacology , Dinoprostone/pharmacology , Protein Kinase C/metabolism , Animals , Callithrix , Cells, Cultured , Cloprostenol/pharmacology , Corpus Luteum/cytology , Corpus Luteum/metabolism , Cyclic AMP/biosynthesis , Dose-Response Relationship, Drug , Female , Progesterone/biosynthesis
8.
J Reprod Fertil ; 97(2): 425-31, 1993 Mar.
Article in English | MEDLINE | ID: mdl-8388957

ABSTRACT

The luteolytic response to a prostaglandin F2 alpha analogue, cloprostenol, was investigated in vivo and in vitro at defined stages of the luteal phase. In vivo administration of cloprostenol to female marmoset monkeys on day 3 after ovulation had no effect on plasma progesterone concentrations, whereas administration on day 14 after ovulation reduced plasma progesterone to preovulatory concentrations within 4 h. To identify the cellular basis for this luteolytic action, marmoset luteal tissue obtained on days 3, 6 and 14 after ovulation was incubated in vitro and progesterone production, cAMP accumulation and phosphoinositide (PI) turnover measured in response to cloprostenol, human chorionic gonadotrophin (hCG) with or without cloprostenol, or dibutyryl-cAMP with or without cloprostenol. Progesterone production was stimulated by both hCG and dbcAMP at all stages of the luteal phase. Although neither hCG nor dbcAMP had any significant effects on PI turnover, hCG also increased cAMP accumulation. In marmoset luteal tissue obtained on day 3 after ovulation, cloprostenol had no significant effect on basal or hCG/dbcAMP-stimulated progesterone production but significantly stimulated PI turnover. In contrast, on days 6 and 14 after ovulation, cloprostenol significantly inhibited hCG- and dbcAMP-stimulated progesterone production and the cAMP response to hCG, but had no significant effect on PI turnover. Since progesterone production by the marmoset corpus luteum depends on the luteotrophic support of luteinizing hormone (LH), these observations suggest that the luteolytic action of cloprostenol in vivo involves the inhibition of LH/hCG action at sites both prior and subsequent to cAMP accumulation. However, such luteolytic effects do not appear to require the generation of inositol phosphates by increased PI turnover.


Subject(s)
Cloprostenol/pharmacology , Corpus Luteum/drug effects , Cyclic AMP/physiology , Inositol Phosphates/physiology , Luteolysis/drug effects , Animals , Bucladesine/pharmacology , Callithrix , Chorionic Gonadotropin/pharmacology , Culture Techniques , Female , Inositol Phosphates/metabolism , Luteal Phase/physiology , Progesterone/blood
9.
Biol Reprod ; 47(4): 554-60, 1992 Oct.
Article in English | MEDLINE | ID: mdl-1391341

ABSTRACT

Although recent data for several species of primate, including human and marmoset, indicate that the corpus luteum secretes high levels of radioimmunoassayable inhibin, the nature of the immunoreactive (ir) inhibin detected has not been established. In this study, plasma ir-inhibin levels during the ovarian cycle of the marmoset (n = 12 animals) were measured by alpha-subunit-directed inhibin RIA, and values were compared with those estimated by a recently developed two-site immunoradiometric assay (IRMA) specific for inhibin alpha-beta dimer. Consistent with earlier data, plasma levels of ir-inhibin measured by RIA (overall mean value 133 +/- 7 ng/ml; n = 171) reached values 4-fold higher (p less than 0.001) during the luteal phase (222 +/- 20 ng/ml) than during the follicular phase (58 +/- 8 ng/ml), being directly correlated with plasma progesterone levels (r = 0.65; p less than 0.001). In contrast, plasma ir-inhibin levels estimated by IRMA were substantially lower than those measured by RIA (overall mean value 9.62 +/- 1.08 ng/ml; n = 171) and did not vary significantly during the cycle. Administration of a luteolytic dose of cloprostenol during the late luteal phase/early pregnancy led to an abrupt fall in plasma concentrations of progesterone (95%) and alpha-inhibin measured by RIA (82%), whereas dimeric inhibin levels remained unchanged. Analysis of marmoset luteal extracts (n = 5) by RIA, IRMA, and inhibin bioassay yielded inhibin estimates of 102.6 +/- 21.0, 0.632 +/- 0.103, and less than 2.0 ng/mg, respectively, thus confirming that only a very small proportion of the inhibin produced was dimeric (i.e., bioactive) in nature.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Callithrix/physiology , Corpus Luteum/metabolism , Inhibins/metabolism , Animals , Cloprostenol/pharmacology , Female , Immunoradiometric Assay , Inhibins/analysis , Inhibins/chemistry , Luteal Phase/drug effects , Luteal Phase/physiology , Menstrual Cycle/drug effects , Menstrual Cycle/physiology , Progesterone/blood , Protein Conformation , Radioimmunoassay
10.
J Reprod Fertil ; 96(1): 385-93, 1992 Sep.
Article in English | MEDLINE | ID: mdl-1432971

ABSTRACT

The role of the embryo in promoting increased plasma concentrations of immunoreactive inhibin after conception in the marmoset monkey was determined by flushing embryos from the uterus between days 5 and 9 after ovulation (implantation commences on days 11-12). Blood samples were taken from each animal (three times a week) after ovulation until the end of the luteal phase. Plasma inhibin concentrations were measured using a radioimmunoassay based on antisera against a synthetic fragment of the alpha-subunit of human inhibin. When embryos were flushed on days 5 and 6 (n = 6) after ovulation inhibin concentrations did not exceed 250 ng ml-1 for the duration of the luteal phase. In contrast when embryos were flushed on days 7 (n = 4), 8 (n = 4) and 9 (n = 3) maximum concentrations of inhibin always exceeded 250 ng ml-1, reaching > 400 ng ml-1 when embryos were flushed on days 8 and 9. Inhibin concentrations remained high for the duration of the luteal phase, which varied in length between 20 and 32 days. Significantly (P < 0.01) higher mean plasma concentrations of immunoreactive inhibin were first recorded on days 7-8 after ovulation in animals that had embryos flushed on days 7, 8 and 9 compared with concentrations in animals that had embryos flushed on days 5 and 6. Inhibin could not be detected in the medium of embryos cultured for up to 2 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Blastocyst/metabolism , Inhibins/biosynthesis , Pregnancy, Animal/metabolism , Animals , Callithrix , Female , Inhibins/blood , Luteal Phase/blood , Pregnancy , Radioimmunoassay
11.
Mol Cell Endocrinol ; 82(2-3): 207-14, 1991 Dec.
Article in English | MEDLINE | ID: mdl-1724435

ABSTRACT

To investigate the involvement of cyclic AMP phosphodiesterase in the antigonadotrophic actions of prostaglandin F2 alpha (PGF2 alpha), human granulosa cells were cultured in serum-supplemented medium for 72 h, treated for 30 min with cloprostenol or phorbol myristate acetate (PMA) and then exposed to human chorionic gonadotrophin (hCG) +/- isobutyl-methylxanthine (IBMX) for a further 4 h. In the absence of IBMX, cloprostenol and PMA inhibited hCG-stimulated progesterone production. However, in the presence of 0.5 mM IBMX, inhibition of phosphodiesterase prevented these antigonadotrophic effects. Phosphodiesterase activity was assessed by a novel direct assay performed on intact cells after 3 days of culture. PGF2 alpha, cloprostenol and 4 beta-PMA all enhanced cAMP degradation whilst an inactive phorbol ester (4 alpha-PMA) had no effect. Down-regulation of protein kinase C by 4 beta-PMA pre-treatment prevented the subsequent stimulation of phosphodiesterase activity by both cloprostenol and 4 beta-PMA. We conclude that the antigonadotrophic actions of PGF2 alpha in cultured human granulosa cells involve a stimulation of cAMP phosphodiesterase mediated via protein kinase C.


Subject(s)
3',5'-Cyclic-AMP Phosphodiesterases/metabolism , Dinoprost/physiology , Granulosa Cells/enzymology , Protein Kinase C/metabolism , 1-Methyl-3-isobutylxanthine/pharmacology , 3',5'-Cyclic-AMP Phosphodiesterases/antagonists & inhibitors , Adult , Cells, Cultured , Cloprostenol/pharmacology , Cyclic AMP/metabolism , Female , Humans , Kinetics , Progesterone/physiology , Tetradecanoylphorbol Acetate/pharmacology
12.
J Endocrinol ; 131(2): 319-25, 1991 Nov.
Article in English | MEDLINE | ID: mdl-1660521

ABSTRACT

The antigonadotrophic action of a prostaglandin F2 alpha analogue, cloprostenol, has been investigated in human granulosa cells obtained from cycles stimulated for in-vitro fertilization and induced to secrete luteal quantities of progesterone by culture in serum-supplemented medium. Cells were exposed to conditions which may mimic those occurring in early pregnancy to establish the roles of human chorionic gonadotrophin (hCG) versus LH and that of cyclic AMP (cAMP) in the anti-gonadotrophic action of cloprostenol. When human granulosa cells were cultured in the absence of treatment for 3 days, exposure to cloprostenol had no effect on basal progesterone production but inhibited hCG-stimulated progesterone (60% decrease; P less than 0.01), hCG-stimulated cAMP (40% decrease; P less than 0.05) and the progesterone response to dibutyryl cAMP (dbcAMP; 70% decrease; P less than 0.01), suggesting pre- and post-cAMP sites of cloprostenol action. The inhibitory actions of cloprostenol were prevented when the granulosa cells were either continuously exposed to treatment from the start of culture or pre-exposed for 3 days to maximum concentrations of LH, hCG, dbcAMP or 8-bromo-cAMP. We conclude that prior exposure either in vivo or in vitro to LH or hCG prevents the subsequent antigonadotrophic action of cloprostenol via a cAMP-dependent mechanism. Prevention of the antigonadotrophic action of cloprostenol after exposure to hCG may be a mechanism through which CG prevents regression of the corpus luteum in early pregnancy, while the suppressive effect of LH pretreatment may account for the refractory response of the early corpus luteum to cloprostenol following the midcycle LH surge.


Subject(s)
Cloprostenol/pharmacology , Cyclic AMP/metabolism , Gonadotropins/pharmacology , Granulosa Cells/metabolism , Luteolysis/physiology , 8-Bromo Cyclic Adenosine Monophosphate/pharmacology , Bucladesine/pharmacology , Cells, Cultured , Chorionic Gonadotropin/pharmacology , Female , Granulosa Cells/drug effects , Humans , Luteinizing Hormone/pharmacology , Progesterone/biosynthesis
13.
J Reprod Fertil ; 92(2): 497-509, 1991 Jul.
Article in English | MEDLINE | ID: mdl-1886105

ABSTRACT

Genes for chorionic gonadotrophin (CG) are transcribed by the 16-cell embryo stage in humans, but there is no clear evidence of CG secretion as a bioactive dimer before attachment and trophoblast outgrowth stages of implantation. The studies summarized question the timing of CG expression and secretion, the possible roles of CG for intraembryonic differentiation and at the implantation site, and the recognition of this primate embryo-derived signal in support of the corpus luteum. The data suggest that the implantation window in primates may be broader than in non-primate species, where a closer synchrony between embryonic, tubal and uterine events appears to be necessary for embryonic survival. Some preliminary data concerning an association between peripheral thrombocytopenia, ovarian inhibin secretion and peri-implantation stages of embryo development indicate that an unknown embryonic signal may be secreted before bioactive CG can be detected.


Subject(s)
Chorionic Gonadotropin/physiology , Embryo Implantation/physiology , Primates/physiology , Animals , Chorionic Gonadotropin/genetics , Chorionic Gonadotropin/metabolism , Corpus Luteum Maintenance/physiology , Embryo, Mammalian/physiology , Female , Gene Expression/physiology , Humans , Inhibins/physiology , Pregnancy
14.
Hum Reprod ; 6(6): 779-82, 1991 Jul.
Article in English | MEDLINE | ID: mdl-1757514

ABSTRACT

Human granulosa cells, from women undergoing ovum collection for in-vitro fertilization (IVF), will luteinize in vitro and provide a model for investigating the antigonadotrophic action of a prostaglandin F2 alpha (PGF2 alpha) analogue, cloprostenol, on granulosa-derived luteal cells. The granulosa cells were cultured in a defined medium and exposed to treatments during a preincubation period of 0 to 3 days and a final incubation with low density lipoprotein (LDL) from days 3 to 4. In the absence of human chorionic gonadotrophin (HCG), progesterone production was low, whereas exposure to HCG in the final incubation resulted in a 10-fold increase in progesterone concentrations. The inclusion of cloprostenol with HCG in the final incubation significantly (P less than 0.05) inhibited HCG-stimulated progesterone production. Exposure to HCG during the preincubation prevented the antigonadotrophic action of cloprostenol in the final incubation. The antigonadotrophic action of cloprostenol was retained when the granulosa cells were exposed to cloprostenol during the preincubation. Omission of LDL from the final incubation lowered the production of progesterone but the pattern of responses to HCG and cloprostenol were similar. Prevention of the antigonadotrophic action of cloprostenol after exposure to HCG may be a mechanism through which chorionic gonadotrophin can prevent regression of the corpus luteum in early pregnancy. Cloprostenol does not appear to inhibit LDL-stimulated steroidogenesis in human granulosa cells.


Subject(s)
Chorionic Gonadotropin/physiology , Cloprostenol/antagonists & inhibitors , Granulosa Cells/drug effects , Progesterone/biosynthesis , Cells, Cultured , Female , Granulosa Cells/metabolism , Humans
15.
J Endocrinol ; 128(3): 465-73, 1991 Mar.
Article in English | MEDLINE | ID: mdl-2013750

ABSTRACT

Peripheral concentrations of immunoreactive (ir) inhibin have been measured during the ovarian cycle and early pregnancy in the marmoset monkey. Blood samples were taken (three per week) during conception (n = 6) and non-conception (n = 5) cycles. Ir-inhibin was measured by radioimmunoassay using an antiserum raised against a synthetic peptide fragment of the alpha subunit of human inhibin. Monomeric bovine alpha subunit and 32 kDa bovine inhibin were used as tracer and standard respectively. In all animals low concentrations of ir-inhibin were recorded during the follicular phase (40-60 micrograms/l) of the cycle. After ovulation, ir-inhibin concentrations increased but the peak concentrations attained differed between conception and non-conception cycles. In non-pregnant animals ir-inhibin concentrations reached a maximum of 242 +/- 16 micrograms/l on days 12/13 after ovulation. In pregnant animals ir-inhibin concentrations were significantly (P less than 0.05) higher (1.8-fold) than in non-pregnant animals on days 8/9 after ovulation, and reached a maximum value of 636 +/- 141 micrograms/l on days 20/21 after ovulation. Administration of an LHRH antagonist during the luteal phase on days 6-8 after ovulation resulted in a significant (P less than 0.05) decrease in progesterone and ir-inhibin concentrations within 4 and 8 h respectively. This was prevented by co-administration with human chorionic gonadotrophin. Administration of cloprostenol to pregnant animals between days 17 and 20 after ovulation halved the initial concentrations of both inhibin and progesterone within 1.5 h.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Cloprostenol/pharmacology , Estrus/blood , Gonadotropin-Releasing Hormone/analogs & derivatives , Gonadotropin-Releasing Hormone/antagonists & inhibitors , Inhibins/blood , Pregnancy, Animal/blood , Animals , Callithrix , Chorionic Gonadotropin/blood , Chorionic Gonadotropin/pharmacology , Female , Gonadotropin-Releasing Hormone/pharmacology , Pregnancy , Progesterone/blood
16.
J Endocrinol ; 128(1): 121-9, 1991 Jan.
Article in English | MEDLINE | ID: mdl-1900320

ABSTRACT

The effects of the luteolytic and luteotrophic agents cloprostenol, human chorionic gonadotrophin (hCG) and melatonin on the corpus luteum have been investigated in marmoset monkeys treated with an LHRH antagonist to reduce endogenous LH secretion. This has allowed the effects of these agents to be investigated in the absence of the principal endogenous luteotrophin. Administration of the LHRH antagonist ([N-acetyl-D beta Nal1-D-pCl-Phe2-D-Phe3-D-Arg6-Phe7-Arg8-D-Ala10]NH2-LHRH) or cloprostenol between days 7 and 11 after ovulation (preimplantation) resulted in luteolysis. A significant (P less than 0.05) decrease in progesterone concentrations had occurred by 4 h after administration of the LHRH antagonist and was indeed preceded by a fall in LH concentrations. Coadministration of hCG with the LHRH antagonist prevented the fall in progesterone. In contrast, administration of cloprostenol resulted in an immediate fall in progesterone concentrations, to less than half the initial level within 1 h, and co-administration with hCG did not prevent the fall. Administration of hCG stimulated progesterone production when given 8 h after the LHRH antagonist but not after 24 h. Cloprostenol prevented the stimulation by hCG. Co-administration of melatonin with the LHRH antagonist did not prevent the decrease in progesterone concentrations. Melatonin was also not effective in preventing the fall in progesterone induced by cloprostenol. However, co-administration of melatonin and cloprostenol between days 17 and 21 after ovulation (post-implantation) significantly (P less than 0.05) delayed the fall in progesterone seen with cloprostenol alone. These results suggest that while the LHRH antagonist and cloprostenol have different sites of action their effect is similar at the corpus luteum, that is in depriving the corpus luteum of luteotrophic support. The results also suggest that melatonin may be able to influence the luteolytic action of cloprostenol but that its effect varies with the stage of the cycle. The physiological role for such an action, if any, remains unknown.


Subject(s)
Cloprostenol/pharmacology , Corpus Luteum/drug effects , Gonadotropin-Releasing Hormone/analogs & derivatives , Gonadotropin-Releasing Hormone/antagonists & inhibitors , Luteal Phase/physiology , Animals , Callitrichinae , Chorionic Gonadotropin/pharmacology , Female , Gonadotropin-Releasing Hormone/pharmacology , Luteinizing Hormone/blood , Melatonin/blood , Melatonin/pharmacology , Progesterone/blood
17.
J Reprod Fertil ; 90(2): 427-37, 1990 Nov.
Article in English | MEDLINE | ID: mdl-2250242

ABSTRACT

The size distribution of marmoset luteal cells was determined on Days 6, 14 and 20 after ovulation in non-pregnant cycles and in early pregnancy. Image analysis was used to estimate the cell diameter of dispersed cells prepared from the marmoset corpus luteum (CL). Steroidogenic cells showed a size distribution consistent with one population of cells. There was a significant increase in mean cell diameter (P less than 0.05) from Day 6 to Day 14 in pregnant and non-pregnant animals with no further increase on Day 20. Micrographs of marmoset luteal tissue showed cells of greater than 10 micron containing the organelles typical of steroid-producing cells, and smaller non-steroidogenic cells surrounding the steroid-producing cells. On the basis of microscopy, there were no areas within the CL where cell composition was noticeably different. In contrast, micrographs of human luteal tissue showed two types of steroidogenic cell; most cells were similar to those in the marmoset CL but a smaller population of smaller cells could be distinguished around the periphery and along vascular septa. It is likely that these smaller and larger types of steroidogenic cells are of theca and granulosa cell origin respectively, the two cell populations differing in the degree of electron density and amount of rough endoplasmic reticulum. A distinguishing feature between marmoset and human luteal cells was the shape of the mitochondrian which were considerably rounder in marmoset luteal cells. The origin of steroidogenic cells in the marmoset CL is unclear, although in marmosets and man the luteal cell types display morphological characteristics distinct from the large and small luteal cells described for CL of the domestic ungulates.


Subject(s)
Callithrix/anatomy & histology , Corpus Luteum/cytology , Pregnancy, Animal/physiology , Animals , Corpus Luteum/ultrastructure , Endoplasmic Reticulum/ultrastructure , Female , Humans , Image Processing, Computer-Assisted , Mitochondria/ultrastructure , Pregnancy , Pregnancy Trimester, First
18.
Br Vet J ; 146(1): 17-23, 1990.
Article in English | MEDLINE | ID: mdl-2306598

ABSTRACT

The effect of prolonged exposure to exogenous melatonin on the reproductive status and growth rate of ewe lambs was investigated. Ewe lambs (born late March) were given intravaginal melatonin implants on 4 July (group J, n = 10) during anoestrus, or 20 December (group D, n = 10) during the breeding season. A third group (group C, n = 7) received empty implants on 4 July. Plasma progesterone concentrations were used to assess reproductive status. In the control group cyclic ovarian activity began on November 15 +/- 4 days, and ceased on January 28 +/- 8 days. In group J the onset of the breeding season was advanced by 4.9 weeks (occurring on October 12 +/- 4 days; P less than 0.001). The onset of anoestrus was also advanced in this group with 9 out of 10 of the ewes ceasing ovarian activity at least 3.4 weeks in advance of the control ewes (P less than 0.01). In contrast the timing of anoestrus was unchanged in group D, occurring on February 14 +/- 6 days. Melatonin treatment of ewe lambs from early July had no effect on growth rate, whereas treatment from mid-December had a depressive effect. Ewes therefore become refractory to the inductive effects of exogenous melatonin after long-term exposure. The results are consistent with the view that photorefractoriness is due to changes in the processing rather than the generation of the melatonin signal.


Subject(s)
Anestrus/drug effects , Estrus/drug effects , Melatonin/pharmacology , Sexual Maturation/drug effects , Sheep/physiology , Animals , Female , Sheep/growth & development , Weight Gain/drug effects
19.
J Reprod Fertil ; 87(1): 301-10, 1989 Sep.
Article in English | MEDLINE | ID: mdl-2560061

ABSTRACT

Dispersed marmoset luteal cells were incubated for 2 h and progesterone production measured after exposure to hCG, cloprostenol, dibutyryl cAMP, PGF-2 alpha, PGF-2, adrenaline or melatonin. The cells were studied on Days 6, 14 and 20 after ovulation in conception and non-conception cycles. Luteal cells from Day 14 non-pregnant marmosets were compared with human luteal cells taken in the mid-luteal phase. All the treatments stimulated progesterone production including cloprostenol, which is luteolytic when administered to the marmoset in vivo, but the degree of response varied with the stage of the cycle or pregnancy and between marmoset and human luteal cells. In the marmoset, overall analysis of the effect of the treatments showed that, on Day 6 after ovulation, there was no significant effect of any of the treatments in cells from pregnant or non-pregnant animals. In contrast, luteal cells from non-pregnant animals on Day 14 showed a significant response to the treatments (F (8,41) = 2.79, P less than 0.0145) whereas cells from pregnant Day-14 animals were responsive; in cells from pregnant animals, the control production of progesterone was high and already equivalent to the levels stimulated by the treatments. By Day 20, cells from pregnant animals produced lower control concentrations of progesterone than did those on Day 14 and there was a significant overall effect of the treatments (F (8,33) = 3.78, P less than 0.003). These results show that the marmoset CL gains responsiveness to treatment between Days 6 and 14 after ovulation in the non-pregnant cycle. In pregnancy, on Day 14, 2 days after attachment of the embryo, the high control concentrations of progesterone and absence of response to treatment suggest that an embryo message may have affected the CL, providing an endogenous stimulus.


Subject(s)
Corpus Luteum Maintenance/drug effects , Corpus Luteum/physiology , Estrus/physiology , Progesterone/biosynthesis , Animals , Bucladesine/pharmacology , Callithrix , Chorionic Gonadotropin/pharmacology , Cloprostenol/pharmacology , Corpus Luteum/drug effects , Dinoprost/pharmacology , Epinephrine/pharmacology , Female , Humans , Melatonin/pharmacology , Pregnancy
20.
Am J Primatol ; 17(1): 73-79, 1989.
Article in English | MEDLINE | ID: mdl-31968856

ABSTRACT

This study describes the concentrations of melatonin in plasma samples taken from marmoset monkeys (Callithrix jacchus) every 4 h over three 24-h periods. A circadian pattern of secretion was apparent, with higher levels recorded at night (20.00-08.00 h) than during the day (08.00-20.00 h) and a peak concentration at 20.00 h. There was a significant difference in the mean day and night concentrations (32.5 ± 4.5 pg/ml versus 49.0 ± 6.9 pg/ml, respectively) with individual concentrations ranging between<10-60 pg/ml in the day and 15-200 pg/ml at night. Circadian plasma melatonin concentrations were similar over the three 24-h periods, in male (n = 3) and female (n = 3) monkeys, and in dominant (cyclic, n = 5) and subordinate (acyclic, n = 4) females. The results show a less pronounced circadian profile in the marmoset than is seen in the human but a similar profile to that in the seasonally breeding rhesus monkey.

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