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Mating stimulates estradiol production by ovaries of the musk shrew (Suncus murinus).

Asian musk shrews (Suncus murinus) are induced ovulators, but exhibit no cyclic changes in reproductive structures or in sexual behavior. Mating behavior is induced by contact with a male. To determine if mating induces changes in ovarian steroidogenesis, ovaries removed from unmated animals and at 3, 10, 15, and 36 h after mating were cultured for 4 h in the presence or absence of gonadotropins (LH + FSH, 1 microgram/ml). Histological analysis revealed no obvious changes in follicular size or appearance at the end of culture in ovaries cultured at 3 and 10 h post-mating, as compared with ovaries from unmated shrews, and mating did not stimulate any discernable changes in steroid secretion in these two groups. However, at the end of the culture period, ovulation had occurred or was occurring in ovaries from 35% of the animals ovariectomized at 15 h after mating, and corpora lutea (CLs) were present in 39% of ovarian pairs obtained 36 h after mating. At 15 h post-mating, ovaries with ovulations secreted three times more estradiol than did ovaries that showed no evidence of stimulation by mating, but there were no differences in testosterone or progesterone production. In contrast, ovaries isolated 36 h post-mating with CLs secreted dramatically more of all three steroids than ovaries without CLs (23, 13, and 52 times more estradiol, testosterone, and progesterone, respectively). These data are consistent with plasma concentrations of estradiol at the time of ovariectomy, which were twice as high at both 15 and 36 h after mating, in animals whose ovaries showed evidence of ovarian stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Abnormal electromyographic activity of the urethral sphincter, voiding dysfunction, and polycystic ovaries: a new syndrome?

A potential association between abnormal electromyographic activity--that is, decelerating bursts and complex repetitive discharges--of the urethral sphincter and difficulty in voiding was examined in 57 women with urinary retention. Abnormal electromyographic activity was found in 33. Ultrasonography of the ovaries in 22 of the 33 women showed that 14 had polycystic ovaries. Of the other eight women, two had had oophorectomies, one had shrunken ovaries and ovarian failure, and one had previously undergone oophorectomy and the other ovary could not be seen; in one neither ovary could be seen, and three had ovaries of normal appearance, although two of these women were taking the contraceptive pill. Thirteen of the group had endocrine symptoms and signs characteristic of the polycystic ovary syndrome. Videocystometrography in 17 of the women who were examined by ultrasonography showed low flow rates and high residual volumes of urine after micturition in 12 women who could void, the other five having chronic urinary retention. A speculative hypothesis for the observed association of impaired voiding, abnormal electromyographic activity of the urinary sphincter, and polycystic ovaries is advanced, based on the relative progesterone deficiency that characterises the polycystic ovary syndrome. Progesterone stabilises membranes, and its depletion might permit ephaptic transmission of impulses between muscle fibres in the muscle of the urethral sphincter, giving rise to the abnormal electromyographic activity. This may impair relaxation of the sphincter, resulting in low flow rates of urine, incomplete emptying of the bladder, and, finally, urinary retention.

Adult↗

The role of inhibin in women with polycystic ovary syndrome (PCOS).

Human follicular fluids from normal women and from women with polycystic ovary syndrome (PCOS) were collected at various stages of the menstrual cycle, and inhibin and steroid contents were determined by rat anterior pituitary cell monolayer bioassay system and specific RIAs, respectively. Inhibin contents in viable follicles were significantly higher than those in atretic and in cystic follicles. Preovulatory and luteal phase follicles had lower inhibin levels than viable follicles in normal women. A significant inverse correlation was found between inhibin contents and androstenedione to estrogen ratios. The inhibin contents in PCOS follicles were not significantly different from those of viable follicles of normal women at the early follicular phase, but they were significantly higher than in atretic follicles. Since PCOS ovaries often contain more antral follicles than normal ovaries, it could be that this potential source of inhibin may be greater than in normal ovaries. Furthermore, 9 out of 10 follicles in any ovary of normal women are atretic and contribute less inhibin production, whereas PCOS ovaries contain many follicles capable of secreting considerable amounts of inhibin. Therefore, PCOS ovaries totally may secrete more inhibin than normal ovaries. This higher potency of PCOS ovaries to secrete inhibin along with higher serum estrogen levels may be responsible for the disparity between basal LH and FSH levels in PCOS.

Animals↗

Between-ovary interaction in the regulation of follicle growth, corpus luteum function, and gonadotropin secretion in the primate ovarian cycle. III. Temporal and spatial dissociation of folliculogenesis and negative feedback regulation of tonic gonadotropin release after luteectomy in rhesus monkeys.

This study was designed to examine effects of previous ovarian status on subsequent follicle growth and the role of between-ovary communication in the regulation of folliculogenesis and gonadotropin secretion during the primate ovarian cycle. Responses to luteectomy were compared in two groups of rhesus monkeys. In the first, follicle growth and corpus luteum function had been constrained chronically to a single ovary by hemiovariectomy performed 66--258 days earlier; the second group was composed of intact monkeys that underwent contralateral wedge resection at luteectomy. In each group, luteal ablation was followed by a prompt fall in serum progesterone levels, a premature onset of menses, and the next preovulatory gonadotropin surges 14.7 +/- 1.1 or 15.4 +/- 1.4 days later (mean +/- SE; P greater than 0.25). Although the patterns of circulating estradiol before and after ablation in each group were superimposable, luteectomy in monkeys lacking a contralateral ovary was followed by a large (2- to 4-fold) and prolonged (7--10 days) increase in serum FSH, whereas in monkeys with two ovaries, serum FSH levels exhibited only a small short-lived rise. The findings indicate that 1) prior chronic constraint of ovarian function to a single ovary did not alter the overall time course of new follicle growth culminating in ovulation after luteectomy; 2) the contralateral ovary provided the principal negative feedback regulation of gonadotropin secretion for some time after luteectomy even though it may not have been the exclusive site of new follicle growth; 3) whereas the ability of the luteectomized ovary to regulate tonic gonadotropin secretion was temporarily impaired, its ability to support the customary temporal pattern of follicle growth after luteal ablation was not; 4) some (contralateral) ovarian factor other than estradiol or progesterone apparently made a major contribution to the regulation of FSH secretion after luteectomy; and 5) folliculogenesis culminating in ovulation from a single follicle and the negative feedback regulation of tonic gonadotropin secretion in some circumstances may occur concurrently but separately on opposite ovaries or may occur at different times within the same ovary.

Animals↗

The effects of a gonadotropin-releasing hormone agonist on ovulation and steroidogenesis during perfusion of rabbit and rat ovaries in vitro.

The ability of a GnRH agonist (GnRHa) to exert direct effects on rat and rabbit ovaries was examined in vitro. Ovaries of estrous rabbits and immature, PMSG-primed rats were surgically removed and perfused with a defined medium via an aortic cannula. In this system, the ovary remains viable and capable of undergoing ovulation in response to LH. Samples of perfusion medium were taken for steroid measurements and the number of ovulations determined by direct observation (rabbit) or oocyte recovery (rat). Follicles of ovaries perfused with medium alone rarely ovulated. GnRHa (0.1 micrograms/ml) induced ovulations in 6 of 7 rat ovaries (4 to 22 ovulations per ovulating ovary) and this effect was blocked by a GnRH antagonist. In contrast, a much higher dose of the agonist (10 micrograms/ml) induced ovulations in only 7 of 15 rabbit ovaries. GnRHa caused small but significant increases in progesterone levels in the perfusion medium in both species in comparison to no treatment. Mean estradiol levels also tended to be higher in the GnRHa groups in comparison to controls but the differences were not significant. GnRHa appears to act directly on both the rabbit and rat ovary but the rat ovary is much more sensitive to its ovulation-inducing effects.

Animals↗

Expression of insulin-like growth factor-binding protein-2 and -3 messenger ribonucleic acid in the porcine ovary: localization and physiological changes.

Insulin-like growth factor-binding protein (IGFBP)-2 and -3 are the most prevalent IGFBPs in porcine follicular fluid, as determined on ligand blots, but little is known about the localization and regulation of their synthesis in vivo. This study was designed to investigate the localization and cyclic regulation of the mRNA for these two IGFBPs in the porcine ovary, RNA was extracted from whole ovaries morphologically classified as immature, preovulatory, and luteal. Northern hybridization analysis of this RNA showed no significant difference in the expression of IGFBP-2 mRNA in these ovaries (OD for preovulatory, luteal, and immature ovaries, 0.076 +/- 0.01, 0.071 +/- 0.01, and 0.10 +/- 0.008/micrograms RNA, respectively). IGFBP-3 mRNA was not different in immature and preovulatory ovaries, but was 10-fold greater (P less than 0.025) in luteal ovaries. Northern analysis of RNA extracted from ovaries also showed no significant change in IGFBP-2 mRNA on days (d) 11, 16, and 21 of the estrous cycle. IGFBP-3 mRNA tended to decrease between d11-16 with the onset of luteal regression and was significantly decreased in d21 preovulatory ovaries to 22% of the values in d11 ovaries. Granulosa, thecal, and luteal cells were also analyzed for IGFBP mRNA. IGFBP-2 mRNA was most abundant in granulosa cells, lower in thecal cells, and lowest in luteal cells. No IGFBP-3 mRNA could be detected in granulosa cells, and luteal cells expressed 15- to 63-fold greater levels than thecal cells. These results show that IGFBP-2 and -3 mRNAs are expressed in specific ovarian cell types and that their expression appears to be independently regulated during the reproductive cycle. This provides further evidence for the importance of these proteins as paracrine/autocrine regulators of ovarian function.

Actins↗

Locally produced angiotensin II induces ovulation by stimulating prostaglandin production in in vitro perfused rabbit ovaries.

The present study was undertaken to investigate the role of exogenous and endogenous angiotensin II (Ang II) in ovarian steroidogenesis and production of prostaglandin (PG) in in vitro perfused rabbit ovaries. The addition of 100 or 10 micrograms Ang II at 2-h intervals to the perfusate did not stimulate progesterone production, but significantly stimulated estradiol (E2) production by perfused rabbit ovaries. When the specific antagonist of Ang II, saralasin at 2 x 10(-6) M, was added to the perfusate 30 min before the onset of Ang II administration, Ang II-stimulated production of E2 was significantly blocked. Ang II also significantly stimulated both PGE2 and PGF2 alpha production, while the addition of saralasin to the perfusate significantly inhibited the Ang II-stimulated production of PG. The levels of PGs in ovaries perfused with saralasin plus 100 micrograms Ang II did not differ significantly from those in control ovaries perfused with medium alone. Exposure to human CG (hCG) significantly stimulated production of progesterone and E2 by perfused rabbit ovaries, while the concomitant administration of 2 x 10(-6) M saralasin significantly reduced only E2 production. Addition of saralasin to the perfusate inhibited hCG-stimulated PG production in a dose-dependent manner. The ovulatory efficiency in ovaries treated with hCG alone or hCG plus saralasin was significantly correlated with PG production by perfused rabbit ovaries at 12 h after exposure to hCG. The production of PG stimulated by Ang II was completely reduced by indomethacin treatment during the entire perfusion period. Indomethacin completely blocked Ang II-induced ovulation, but not Ang II-stimulated oocyte maturation. Concurrent administration of staurosporine, a protein kinase C inhibitor, at 10(-6) M significantly inhibited Ang II-stimulated meiotic maturation of ovulated ova and follicular oocytes. In conclusion, these results indicate that Ang II has a direct role in ovarian production of E2 and PG. An intrinsic renin-angiotensin system in the rabbit ovary may act as an intermediary of gonadotropin-stimulated PG production. Locally produced Ang II may induce ovulation in the rabbit ovary, at least in part, by stimulating PG production.

Angiotensin II↗

Granulosa cell production of anti-Müllerian hormone is increased in polycystic ovaries.

CONTEXT: There has been renewed interest in anti-Müllerian hormone (AMH) because of its role in the ovary. Data on its actions are sparse, but it appears to inhibit follicle growth. Interestingly, serum AMH is two to three times higher in women with polycystic ovary (PCO) syndrome than women with normal ovaries. OBJECTIVE: We examined the production of AMH by cells from a range of follicle sizes from normal ovaries and compared this with production by ovulatory and anovulatory (anov) PCOs. DESIGN: Granulosa cells (GCs) and theca and follicular fluid (ff) were isolated from intact follicles. Cells were cultured for 48 h +/- FSH or LH, and AMH was measured in ff and cell-conditioned media (CM). RESULTS: AMH levels in ff and GC-CM ranged from 42 to 2240 and 0.025 to 1.7 ng/ml, respectively, and were low or undetectable in ff and GC-CM from follicles greater than 9 mm, luteinized cells, and theca and stroma. The mean level of AMH was four times higher in GC-CM from ovulatory PCOs [mean (range) 1.56 (0.025-7)] and 75 times higher from anovPCO [21.4 (17.2-43 ng/ml)] than normal ovaries [0.37 (0.025-1.7)]. Neither LH nor FSH had an effect on AMH production by GCs from normal ovaries, but in cells from PCOs, FSH significantly decreased AMH, and in contrast, LH increased AMH. CONCLUSIONS: The reduction of AMH in follicles greater than 9 mm from normal ovaries appears to be an important requirement for the selection of the dominant follicle. AMH production per GC was 75 times higher in anovPCOs, compared with normal ovaries. This increase in AMH may contribute to failure of follicle growth and ovulation seen in polycystic ovary syndrome.

Adult↗

Development and fertility of ovaries in the B6.YDOM sex-reversed female mouse.

When the Y chromosome of Mus musculus domesticus (YDOM) was introduced onto the C57BL/6 (B6) mouse background, half of the XY progeny (B6.YDOM) developed bilateral ovaries and female internal and external genitalia. We examined the fertility of the B6.YDOM sex-reversed female mouse. The chromosomal sex of the individual mouse was identified by dot hybridization with mouse Y chromosome-specific DNA probes. The results indicated that all XY females lacked regular estrous cyclicity although most were able to mate and ovulate after treatment with gonadotropins. When they had been ovariectomized and grafted with ovaries from the XX female litter mate, they initiated estrous cyclicity. Reciprocally, the XX female that had received XY ovarian grafts did not resume estrous cyclicity. Development of the XY ovary was morphologically comparable to the XX ovary until 16 day of gestation (d.g.), when most germ cells had reached the zygotene or pachytene stage of meiotic prophase. However, by the day of delivery (19 or 20 d.g.), no oocyte remained in the medullary cords of the XY ovary. In the control XX ovary, the first generation of follicles developed in the medullary region, and 5 delta-3 beta-hydroxysteroid dehydrogenase (3 beta-HSDH) activity appeared first in the stromal cells around growing follicles by 10 days after birth. In contrast, in the XY ovary, follicles were not formed in the medullary region, and 3 beta-HSDH activity appeared in epithelial cells of the oocyte-free medullary cords. Primordial follicles in the cortex region continued development in both the XX and XY ovaries. These results suggest that the XY female is infertile due to a defect inside the XY ovary. The prenatal loss of oocytes in the medullary cords may be a key event leading to abnormal endocrine function, and thereby, the absence of estrous cyclicity.

Animals↗

Gonadal sex reversal of the developing marsupial ovary in vivo and in vitro.

Undifferentiated tammar wallaby ovaries were transplanted under the skin of male pouch young during the period of mitotic division of the XX germ cells. After 25 days, all the germ cells had disappeared and the ovaries contained seminiferous-like cords. Similarly, undifferentiated ovaries cultured for 4 days with recombinant human Müllerian-inhibiting substance (rhMIS) also contained well-differentiated seminiferous-like cords and few or no surviving germ cells. The majority of controls cultured without rhMIS developed as normal ovaries. However, in a few control ovaries seminiferous-like cords developed in those regions of the ovaries that were partially necrotic and contained few germ cells. These results strongly suggest that sex-reversal of the tammar ovary is the direct result of loss of mitotic germ cells, rather than an effect of MIS on female somatic cells. MIS is apparently toxic to these female germ cells in mitosis, but not to male germ cells in mitosis. Thus, in normal development in the tammar, the presence of XX germ cells in the ovary inhibits the formation of seminiferous cords so that the gonad develops as an ovary.

Animals↗

Blood flow in the ovary and adjacent structures of the non-pregnant sheep.

Blood flow of the ovary, vascular pedicle and oviduct was measured in anaesthetized non-cycling and cycling ewes by timed collection of ovarian venous blood. The degree of arterio-venous shunting across the ovary and pedicle was estimated both in vivo and in vitro by perfusing the tissues with 15 +/- 5 micrometers radioactive microspheres. The mean ovarian blood flow in non-cycling animals was 1.9 ml/min, which was 51% of blood flow in the ovarian vein. In cycling animals ovarian blood flow at midcycle was 2.9 ml/min (66% of ovarian venous flow) in non-luteal ovaries and 4.3 ml/min (79% of venous flow) in luteal ovaries. The degree of arterio-venous shunting was low in all stages of the cycle (1.0-2.6% across ovary + pedicle). The degree of shunting was also found to be very small in vitro (0.007-1.38%) in both non-luteal and luteal ovaries. A considerable number of microspheres was entrapped in the vascular pedicle of the ovary indicating the presence of an extensive capillary bed. There was an inverse relationship between blood flow in the ovary and flow in the vascular pedicle. Alterations in distribution of blood flow between the ovary and adjacent structures supplied by the ovarian artery may be of functional significance in allowing rapid changes in ovarian blood flow. The results of the present study indicate that changes in ovarian blood flow during the oestrus cycle are not caused by an action on arterio-venous shunt vessels.

Animals↗

Role of thyroid hormone in controlling the concentration of luteinizing hormone/human chorionic gonadotropin receptors in rat ovaries.

Little is known about the mechanism by which thyroid hormones influence ovarian function, especially in gonadotropin receptor formation. In this study the concentration of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptors in the ovaries of hypo- and hyperthyroid rats was estimated. Rats were made experimentally hypothyroid by thyroidectomy (N = 10) and hyperthyroid by injections of 40 micrograms of L-thyroxine daily for 21 days (N = 14). After 3 weeks the ovaries were excised, weighed, immersed in liquid nitrogen and then, after 24 h of incubation with 125I-labeled hCG (CR-121), the concentration of receptors (cpm) for one ovary and 1 mg of tissue was counted in their respective homogenates. The ovaries of the hyperthyroid group were diminished in size and consequently the level of receptors per ovary also was reduced when compared with control animals. The number of receptors per ovary and per milligram of tissue of hypothyroid rats was three times higher than in the control. In hyperthyroid animals a significant decrease in these values was noted when compared with hypothyroid rats, especially in the calculation of receptor concentration per ovary. It may be concluded that thyroid function may affect the size of the gland and also the number of LH/hCG-binding sites in rat ovaries. These data may be useful for interpretation of the pathophysiology of polycystic ovary syndrome in women and animals.

Animals↗

Structural and functional changes in ovaries from adult mice treated with diethylstilboestrol in the neonatal period.

Ovaries from 8-week-old female NMRI mice in different stages of the oestrous cycle, or from females neonatally treated with the synthetic oestrogen diethylstilboestrol (DES; 5-10(-6) micrograms daily for 5 days), were studied histologically and for the ability to synthesize steroids from [3H]pregnenolone in vitro. Daily doses of 10(-4) micrograms DES or higher resulted in absence of corpora lutea. In ovaries lacking corpora lutea, the interstitial tissue dominated and the cells in this compartment were large with a clear cytoplasm. The steroids synthesized in ovarian homogenates were separated with thin-layer chromatography. The homogeneity of the steroids was checked in recrystallization experiments. Daily doses of 5-10(-4) micrograms DES in the neonatal period resulted in pronounced deviations in the pattern of ovarian steroids synthesized as compared with control ovaries. In DES-exposed ovaries, the synthesis of androstenedione and, above all, progesterone was high while the synthesis of 17 alpha-hydroxyprogesterone and testosterone was reduced compared with controls. These results could argue for a difference in activities of 17 alpha-hydroxylase and 17 beta-ol-dehydrogenase in ovaries from DES-treated females compared with controls. After transplantation of DES-exposed ovaries to ovariectomized control females, the steroid pattern changed to that typical for control ovaries. Control ovaries transplanted to DES-treated females had a steroid pattern similar to that of DES-exposed ovaries.

17-alpha-Hydroxyprogesterone↗

Methodology and characterization of an in vitro perfusion model for the mouse ovary.

Perfusion of intact ovaries in vitro has proved to be a suitable model for the study of ovarian events, such as ovulation and corpus luteum function, with advantages over cell culture systems of preserved three-dimensional structure with intact intercellular communication. This methodology has been described for several larger experimental animals. Development of genetically manipulated mouse strains and the larger availability of recombinant proteins and monoclonal antibodies in this species prompted the development of a mouse ovary perfusion system. In the present study, we describe the methodology for perfusion of mouse ovaries in vitro and characterize the model with respect to steroidogenesis, ovulatory efficiency and morphology. Swiss mice (30-45 g) were synchronized with a single injection of a GnRH agonist and laparotomy was performed on the morning of pro-oestrus. The right ovary and its vasculature, after cannulation on the arterial side, were surgically isolated and transferred to a recirculating perfusion system, and were perfused for 20 h. The ultrastructure of the ovaries after 20 h of perfusion was examined by transmission electron microscopy, which showed well-preserved cellular organelles and nuclei and distinct tight junctions between endothelial cells. Unstimulated ovaries did not ovulate and secreted small quantities of progesterone. After addition of hCG (50 iu) or ovine LH (0.1 microgram ml-1), ovulations were observed in all perfused ovaries (2.6 +/- 0.5 and 2.0 +/- 0.3 ovulations per treated ovary, respectively). There was a distinct rise in progesterone output after hormone addition. When a phosphodiesterase inhibitor, isobutylmethylxanthine (0.2 mumol l-1), was added in combination with LH, the ovulation rate (7.4 +/- 1.2) was similar to that in vivo and progesterone secretion was further increased compared with other treatments. Since the mouse has become a very important experimental animal in medical research and we have the ability to genetically manipulate this species, this methodology for perfusing mouse ovaries in vitro may be a useful tool in future studies of ovarian physiology.

1-Methyl-3-isobutylxanthine↗

Differential effects of ovarian local anaesthesia during pro-oestrus on ovulation by the right or left ovary in normal and hemi-ovariectomized adult rats.

The effects on spontaneous ovulation in normal and hemi-ovariectomized cyclic rats of an injection of saline or a local anaesthetic (2-diethylamine-2',6'-acetoxilidide hydrochloride with adrenaline, Xylocaine; administered at 13:00 h) into the bursa ovarica, were studied to investigate the participation of ovarian innervation in the regulation of the response of ovarian follicles to endogenous gonadotrophins on the day of pro-oestrus. The injection of saline or Xylocaine into both ovaries did not affect ovulation by the right ovary. Bilateral injection of Xylocaine did not affect ovulation by the left ovary. However, when saline was injected into the right ovary and Xylocaine into the left ovary, the number of ova shed by the left ovary decreased significantly (0.9 +/- 0.7 versus 4.4 +/- 0.2, P < 0.01; Mann-Whitney U test). In hemi-ovariectomized animals, when the right ovary was anaesthetized, ovulation was similar to that in saline-treated animals, and when the right ovary was anaesthetized, ovulation was blocked (one of six animals ovulated). The injection of hCG at 14:00 h to hemi-ovariectomized rats did not restore ovulation, whereas all the rats ovulated when anaesthesia occurred at 18:00 h. The present results suggest that on the afternoon of the day of pro-oestrus, before the LH peak, a neural signal modulates the response of the left ovary to gonadotrophins.

Anesthetics, Local↗

Growth hormone receptors in ovary and liver during gametogenesis in female rainbow trout (Oncorhynchus mykiss).

Changes of growth hormone receptivity in the ovary during the reproductive cycle were studied in rainbow trout (Oncorhynchus mykiss). A method for characterizing growth hormone receptors in crude ovary homogenate was required for this. Binding of radiolabelled recombinant rainbow trout growth hormone (125I-labelled rtGH) to crude ovary preparation was dependent on ovarian tissue concentration. The sites were specific to growth hormone, with no affinity for prolactins and gonadotrophins. Similar high affinities for 125I-labelled rtGH were obtained with crude ovary (4.2 x 10(9) +/- 0.3 mol l-1) and crude liver preparations (4.9 x 10(9) +/- 0.1 mol l-1) at all stages of ovogenesis, and with ovarian membrane preparations (8.2 x 10(9) mol l-1) tested at the beginning of vitellogenesis. Ovarian growth hormone receptor concentration was highest during the early phases of follicular development (endogenous vitellogenesis: 315-310 fmol g-1 ovary) and decreased regularly during oocyte and follicular growth (exogenous vitellogenesis) to reach a minimal value at oocyte maturation (42 fmol g-1 ovary). In postovulated fish, binding was at a similar level (297 fmol g-1 ovary) to that found in endogenous vitellogenesis. Conversely, the absolute binding capacity of the whole ovary was low from immaturity to early exogenous vitellogenesis (0.1-0.6 pmol per pair of gonads), increased slowly during vitellogenesis and more markedly during rapid oocyte growth and at the time of final maturation (10.8 pmol per pair of gonads). In postovulated fish, the absolute binding capacity decreased partially (4.4 pmol per pair of gonads). Mean hepatic growth hormone receptor concentration did not vary with the reproductive stage for most of the cycle (3.0-4.5 pmol g-1 liver) except in endogenous vitellogenesis where significantly higher concentrations were observed (6.7 pmol g-1 liver). Individual ovarian growth hormone receptor concentrations were correlated with hepatic growth hormone receptor concentrations, indicating that they are regulated in a similar way. We conclude that growth hormone receptors are present in the ovary during the entire ovarian cycle in rainbow trout, probably mainly in somatic cells as indicated by the same concentration of binding sites in immature and in postovulated fish. Growth hormone is potentially important during oocyte recruitment in vitellogenesis and initiation of growth and during final follicular maturation.

Analysis of Variance↗

Early postnatal methoxychlor exposure inhibits folliculogenesis and stimulates anti-Mullerian hormone production in the 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 microM) 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 microM 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 production in the ovary. This novel finding suggests that elevated AMH may play a role in MXC's inhibitory effect in the ovary.

Animals↗

Surgical transposition of the ovaries: imaging findings in 14 patients.

Pelvic radiation therapy for cervical or vaginal cancer often leads to ovarian failure. To remove the ovaries from the radiation portal and preserve their function, they can be transposed to the lateral abdomen. Serial imaging studies in 14 patients who had undergone ovarian transposition (five bilateral, nine unilateral) were reviewed. Images obtained included 32 CT scans, 20 sonograms, and one MR image. Most transposed ovaries were located along the paracolic gutters near the iliac crests, creating an extrinsic mass effect on adjacent bowel. Detection of surgical clips on the ovary on CT scans allowed confident recognition of all 19 transposed ovaries. Cysts in the transposed ovaries, noted on most imaging studies, did not correlate with complications of pain or hormonal dysfunction. In one case, a large physiologic cyst in a transposed ovary distorted the cecum and was mistaken for a mucocele of the appendix. In another case, a large ovarian cyst was thought to be tumor recurrence or a lymphocele. These findings indicate that although the transposed ovaries can be recognized on CT scans by the surgical clips attached to the ovaries, the appearance of the ovary does not predict reliably the development of complications.

Adult↗