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Characterization of oestrus and timed collection of oocytes in the grey short-tailed opossum, Monodelphis domestica.

A system of test-pairing was used to detect reproductive behaviour in the grey short-tailed opossum. This enabled timing and characterization of the development of pro-oestrous and oestrous behaviour, and facilitated collection of ovulated and unovulated oocytes. Oestrus was induced 8.5 days (n = 80, 95% confidence limits 7.56-9.21) after the introduction of a male. Timed examination of the ovaries by laparotomy indicated that ovulation occurred 14-16 h after the first onset of oestrous behaviour. The development of follicles was linked to pro-oestrous behaviour, and ovulation occurred in the absence of copulation. Vaginal exfoliative cytology indicated that pro-oestrous behaviour was associated with an increasing number of keratinized epithelial cells, and at the time of maximum receptivity to males, a heavy infiltration of polymorphonuclear leucocytes was seen. Oocytes were typically marsupial: large (approximately 250 microns in diameter), with a yolky vitellus and thin zona pellucida. An average of 6 oocytes were ovulated per ovary.

Animals↗

Follicular, endocrine and behavioural dynamics of the Indian rhinoceros (Rhinoceros unicornis) oestrous cycle.

Longitudinal ultrasound, behaviour and endocrine evaluations were conducted, over 14 to 18 months, in two young female Indian rhinoceroses (Rhinoceros unicornis) to characterize the oestrous cycle. Both females showed the same pattern of follicular development producing a large follicle (10-12 cm diameter) on one of the ovaries that persisted for 8.5 +/- 4.68 days before spontaneously ovulating. Ovulation occurred in all eight cycles monitored in a 6- to 7-year-old female versus 10 out of 14 cycles monitored in a 5- to 6-year-old female. Ultrasound examinations confirmed follicular collapse 48 h following the onset of behavioural oestrus in ovulatory cycles, while anovulatory cycles were associated with the formation of a haemorrhagic follicle. The day of behavioural oestrus corresponded to peak urinary oestrogen conjugate concentrations for each cycle, but anovulatory cycles had lower concentrations on the day of behavioural oestrus compared with ovulatory cycles. A transient increase in urinary progesterone metabolite concentrations was detected 1 day prior to ovulation. Irregular urinary progesterone metabolite profiles followed anovulatory cycles, reflecting varying degrees of follicular luteinization. In an attempt to ensure that a cycle would result in ovulation in the 5- to 6-year-old female, a GnRH treatment was tested during two separate cycles. Administration of GnRH on the day of behavioural oestrus resulted in an increase in urinary luteinizing hormone concentrations 2 h following injection. Regardless, ovulation did not occur in response to treatment. This study provides the first ultrasound data on ovarian activity in the Indian rhinoceros and establishes normal physiologic and behavioural relationships during the oestrous cycle that may facilitate the breeding of this species in captivity.

Animals↗

Tissue inhibitor of metalloproteinase-1 (TIMP-1) deficient mice display reduced serum progesterone levels during corpus luteum development.

TIMP-1 is a multifunctional protein that is abundantly expressed within the ovary during the period of early corpus luteum formation. TIMP-1 has been suggested to play a role in progesterone production, but reports are conflicting. To more thoroughly examine the role of TIMP-1 in steroidogenesis during early luteal development in vivo, immature wild-type and TIMP-1 null female mice were primed with gonadotropins. Mice were sacrificed at 0, 24, and 48 h post hCG administration and serum was collected for determination of estradiol (E(2)) or progesterone (P(4)) concentrations. In the 24 h hCG groups, ovulation was assessed by counting the number of shed ova. Serum P(4) concentrations were significantly lower in the TIMP-1 null mice, both at 24 and 48-h post hCG compared to wild-type counterparts. No differences were detected in the number of ovulations between genotypes at the 24 h time point. In both the 24 and 48 h post-hCG groups, null mice had significantly higher ovarian wet weights. Interestingly, ovarian MMP activity was greater in the null mice at 24 h post hCG but higher in the wild-type at 48 h post hCG administration. These observations suggests that expression of TIMP-1 during early luteal development may participate in regulation of progesterone production via its ability to regulate ovarian MMP activity.

Animals↗

Ultrasound in the timing of artificial insemination with frozen donor semen.

Ultrasonography was performed to time ovulation in 30 patients participating in program for artificial insemination with donor semen (AID). During this study (10 months) 61 scans were carried out where one or more follicles were visible. The mean number of treatment cycles per patient was 2.8. Insemination with frozen donor semen was performed if a clear and mature follicle was detectable. The mean follicular diameter in successful inseminations was 20.1 +/- 0.8 mm, ranging from 16 to 23 mm. Twelve patients became pregnant, 9 (75%) of them after three treatment cycles. The mean time to conception was 2.75 cycles and the most successful day of the menstrual cycle was day 13. Ovulation was induced in 8 (56.7%) patients who conceived. Ultrasound is a noninvasive, quick and reliable method in the detection and timing of ovulation in most AID patients.

Adult↗

Infertility in women: diagnostic evaluation with hysterosalpingography and other imaging techniques.

This article examines the current roles of hysterosalpingography and other imaging studies used in diagnostic assessment and treatment of infertility in women. The advantages and limitations of hysterosalpingography, transabdominal or transvaginal sonography, and MR imaging in detecting abnormalities of the uterus (congenital anomalies, intrauterine adhesions, myometrial and endometrial derangements) and tubal and peritubal disease are presented. The use of sonography to evaluate cyclical endometrial changes, to assess development of the follicle and corpus luteum, to determine whether ovulation has occurred, and to detect dysfunctional ovulatory dynamics is described, as is the role of this technique in improving the prognostic value of the postcoital test for assessing the quality of cervical mucus. With the development and availability of newer and improved methods for the treatment of infertility disorders in women, comprehensive and detailed evaluation to determine the precise underlying cause or causes of the infertility is important to allow formulation of the most appropriate therapeutic plan. With careful and thorough assessment, the cause of infertility can be identified in 90-95% of couples evaluated.

Adult↗

[Studies on the determination of optimal insemination time using electrometric resistance measurements in heifers and young cows after estrus synchronization].

Studies were conducted on how to use under the conditions of industrialised production in the GDR an electrometric method proposed by Soviet scientists to determine oestrus and ovulation date in oestric-synchronised heifers and cows. "Oestrometer-2", an instrument developed in the USSR, was used for 4,818 measurements of electric resistance in 700 oestric-synchronised heifers and 39 oestric-synchronised cows. The typical course of the resistence curve was confirmed by the results. Individual data variation together with unsatisfactory data reliability of the instrument, when applied to measurement at vaginovestibular mucosa, were likely to suggest that the method could not be used with good effect for high-accuracy determination of the ovulation date and therefore, was not useful in helping to save semen pellets after oestric synchronisation.

Animals↗

Incidence of spontaneous ovulation in young, group-housed cats based on serum and faecal concentrations of progesterone.

Cats are considered to be reflex ovulators that exhibit a luteal phase (pregnancy or pseudopregnancy) only after copulatory stimulation. In a group-housed colony of 15 1-year-old domestic queens, 23 noncopulatory, spontaneous ovulations were observed in 87% of the queens over 4.5 months based upon the detection of increased concentrations of progesterone in faeces, serum, or both. Luteal phases were detectable for periods of 3 to 5 weeks with peak progesterone concentrations averaging 21 +/- 1 ng ml-1 in serum and 1874 +/- 281 ng g-1 in faeces. Individual cats exhibited from 0 to 3 spontaneous ovulations and subsequent pseudopregnancies each. A male was added to a separate cage within the room during the last 1.5 months of the study. The incidence of ovulation per 10 day period ranged from 0% to 22% before introduction of the male and from 33% to 57% immediately after introduction of the male, suggesting a potential noncopulatory influence of the male on the incidence of spontaneous ovulation in young, group-housed cats.

Animals↗

Follicular and FSH dynamics in ewes with a history of high and low ovulation rates.

Daily transrectal ultrasound scanning and twice-daily blood sampling were used to monitor the temporal relationships between FSH concentrations and follicle development during complete interovulatory intervals for ewes in which the ovulation rate in each of the 2 previous years was high or low (> or = 3 and < or = 2 ovulations, respectively). Follicles that reached > or = 5 mm were used to define a follicular wave and were tracked retrospectively to 3 mm (emergence). The hypothesis that FSH surges (identified with a computer program) and follicular waves (retrospectively determined based on ultrasound scanning) are temporally associated was supported in both groups by the emergence of an anovulatory or ovulatory follicular wave near the peak of an FSH surge. Further support for the hypothesis was a significant increase in FSH concentrations before and a significant decrease after follicular-wave emergence in both groups independent of the identification of FSH surges. Ewes with a history of high ovulation rates had smaller follicles (anovulatory and ovulatory) and more ovulations, but the 2 groups were similar in the number of ovulatory follicular waves and associated FSH surges, number and characteristics of the FSH surges, and mean FSH concentrations per interovulatory interval. Surges of FSH were periodic (every 3 or 4 d) regardless of the ovulation-rate group or follicle response. In ewes with a low ovulation rate, the nonovulatory FSH surges were most frequently associated with emergence of detected anovulatory follicular waves. In ewes with a high ovulation rate, more FSH surges were not associated with a detected follicular wave, as defined, presumably because the largest follicle did not reach 5 mm. The results indicated that the factors resulting in a high ovulation rate were not exerted through circulatory patterns or concentrations of FSH but involved a shorter growth phase and smaller maximal diameter of follicles.

Animals↗

An objective method for detecting the shift in basal body temperature in women.

The upward shift in basal body temperature (BBT) observed in women of child-bearing age is often used as an indicator that ovulation has occurred and that a period of infertility has begun. In natural family planning programmes it is important to detect the BBT shift reliably and with a minimum of user uncertainty, against a background of physiologically induced temperature variations. The cumulative sum (CUSUM) test is adapted to the problem of detecting BBT shift and applied to 137 BBT charts obtained from 21 women using natural family planning. Results show successful detection of the shift in all cases. Shifts occurred at gynaecologically appropriate times in the menstrual cycle.

Analysis of Variance↗

Differential expression of glycoside residues in the mammalian zona pellucida.

The mammalian zona pellucida is an extracellular matrix surrounding the oocyte, and is composed of three major glycoproteins, ZP1, ZP2, and ZP3. Previous studies have suggested that the sperm receptor activity of the zona pellucida resides in specific oligosaccharide chains on the ZP3 glycoprotein. However, the nature of the terminal monosaccharide(s) on these glycosidic chains to which sperm bind is a matter of active debate. Evidence has been presented to support a role for at least three distinct monosaccharides in sperm binding, alpha-galactose, L-fucose on Lewis X structures, and beta-N-acetylglucosamine. Previous studies have shown that beta-N-acetylglucosamine is uniformly distributed throughout the zona matrix. In this study, we have investigated the expression and distribution of alpha-galactose and fucose moieties during the maturation of the zona pellucida in mouse, rat, and hamster. Interestingly, alpha-galactose residues are expressed only during later stages of zona secretion and, consequently, are confined to the inner portions of the mature zona pellucida in mouse and rat. In hamster, alpha-galactose residues are only detectable in the zona pellucida of ovulated eggs, and are not found in ovarian oocytes. Fucosyl residues linked to Lewis X glycosides are not detectable at any stage of zona maturation in these three species, whereas fucose linked to N-linked core oligosaccharides are present throughout the zona. These studies indicate a previously unappreciated heterogeneity in the composition of zona glycosides. The specific localization of alpha-galactose residues to the inner portions of the zona matrix suggest a role in the later stages of sperm penetration through the zona. Finally, due to their absence from the zona surface, alpha-galactose and Lewis X fucosyl residues are not likely to be mediators of primary sperm binding.

Animals↗

A role for trkA nerve growth factor receptors in mammalian ovulation.

Several members of the neurotrophin (NT) family, including nerve growth factor (NGF), NT-3, and NT-4/5, are expressed in the mammalian ovary. As their respective receptor tyrosine kinases are also found in the gland, the possibility exists that NTs act directly on the gonads to exert effects unrelated to their support of the ovarian innervation. We now report that trkA, the NGF receptor tyrosine kinase, is involved in the acute activational process that leads to the first ovulation. The trkA gene becomes transiently expressed in periovulatory follicules at the time of the first preovulatory surge of gonadotropins at puberty; the increase in trkA messenger RNA (mRNA) content is dramatic ( > 100-fold), but transient (approximately 9 h). No such changes in trkB or trkC mRNA were observed; the abundance of these mRNAs, which encode the receptor tyrosine kinase for NT-4/5 and brain-derived neurotrophic factor, and NT-3, respectively, remained at very low levels throughout puberty. In vivo and in vitro experiments demonstrated that the activation of trkA gene expression is brought about by the proestrous discharge of LH. The increase in trkA mRNA levels is mainly localised to cells of the follicular wall and interstitial tissue of the ovary. NGF mRNA abundance also increases at proestrus, with peak values detected about 5 h before ovulation; as in the case of trkA mRNA, NGF mRNA was found in thecal-interstitial cells. Both trkA and NGF protein, detected by immunohistochemistry, were localized to this same ovarian compartment. Interleukin-1 beta (IL-1 beta), a putative mediator of LH action, enhances both trkA and NGF gene expression in ovarian cells, an effect prevented by IL-1ra, a natural IL-1 beta receptor antagonist. Il-1 beta also stimulates PGE2 release, and this effect was inhibited by both NGF antibodies and a trk receptor blocker, NGF antibodies administered in vivo attenuated the increase in ovarian PGE2 synthesis that antedates ovulation. Immunoneutralization of NGF action or pharmacological blockade of trk tyrosine kinase activity targeted to one ovary resulted in the ipsilateral inhibition of ovulation. The remarkably narrow time frame of trkA gene activation at the completion of follicular growth suggests that NGF acting as a neuroendocrinotrophic factor in a developmentally restricted manner contributes to the acute cytodifferentiation process that leads to the first ovulation in mammals.

Animals↗

Luteinized unruptured follicle syndrome: hormone assay on peritoneal fluid in a laparoscopic study of infertile women.

33 infertile women with normal ovulatory cycles were investigated for the presence of a Luteinized Unruptured Follicle Syndrome (L.U.F.) using steroid hormone assays in peritoneal fluid and laparoscopic visualization of ovulation stigmata. We failed to identify a stigma in 36% (12) of the patients in the early luteal phase, 1 subject had a cystic corpus luteum and in 4 cases no diagnosis was made due to the presence of adhesions. The mean hormone concentrations in PF were significantly higher when the stigma was present (17-beta-estradiol, P less than 0,05; progesterone, P less than 0,01; 17-oh-progesterone, P less than 0,05). The two groups (with and without ovulation stigmata) showed no differences in plasma levels of Estradiol (E2) and Progesterone (P). Stigmata were detected only in 17% of subjects with concomitant endometriosis. 3 patients with a luteal phase defect showed low levels of steroids in PF in spite of the presence of an ovulation stigma.

17-alpha-Hydroxyprogesterone↗

Distinct roles for the mammalian A-type cyclins during oogenesis.

There are two A-type cyclins in higher vertebrates, cyclin A1 and A2. Targeted mutagenesis has shown that cyclin A2 is essential for early embryonic development while cyclin A1 is required only for male meiosis. The embryonic lethality of cyclin A2 knockout mice has obviated understanding its role in other aspects of mammalian development, including the germ line. We reported previously that cyclin A2 expression in the male germ line is consistent with a role in both mitotic and meiotic cell cycles. Using in situ hybridization and immunohistochemistry, we now observe high levels of cyclin A2 in granulosa cells and less-abundant but readily detectable expression in ovarian and ovulated oocytes. A decrease in cyclin A2 protein was observed in oocytes from embryonic stages to post-natal and adult ovaries. Interestingly, cyclin A2 protein was nuclear in oocytes from embryonic day 13.5 to 15.5, changing to largely cytoplasmic in oocytes from embryonic day 16.5 to post-natal and adults. Readily detectable expression of the cyclin-dependent kinases Cdk1 and Cdk2, two common partners for the A-type cyclins, was observed in granulosa cells and oocytes at all stages of folliculogenesis. Cdk1 was predominantly cytoplasmic, whereas Cdk2 was both cytoplasmic and nuclear in oocytes. No cyclin A1 expression, at either the mRNA level or the protein level was detected in either embryonic or adult ovaries, consistent with the full fertility observed in female cyclin A1-deficient mice. These results suggest that in the female germ line, cyclin A2 but not cyclin A1 has distinct roles in both mitosis and meiosis.

Animals↗

Effects of boar contact and housing conditions on estrus expression in sows.

In this paper, the authors review the effects of boar contact and different components of boar presence on onset and expression of estrus in weaned sows. Evidence is presented that boar contact may influence LH release, onset of follicle development and timing of ovulation after weaning. Once the sow is in estrus it is important that she shows estrous behavior, because her expression of estrus determines whether she will be inseminated. Boar contact or components thereof affect expression of estrus in sows. There are distinct differences between different components of boar contact in their effectiveness in the induction of estrous behavior (standing response) in sows. Habituation to boars (due to frequency of boar contact or housing of boars near sows) also affects estrus expression in sows. It is important to inseminate sows at the correct moment relative to ovulation. The use of different estrus detection protocols (e.g., by giving sows different levels of boar stimuli during estrus detection) may result in the definition of distinct periods of estrus that may help to predict the moment of ovulation. However, results to date are not very encouraging. Besides boar stimuli, the housing conditions of sows may affect onset of estrus and estrus expression. This paper focuses on social sow-to-sow interactions. The effects of group housing (as compared to individual housing) on onset of estrus and expression of estrus are equivocal. These effects likely depend on factors like aggression between pen mates, reproductive status of pen mates and social rank of sows within the group.

Animals↗

Gonadotropin-releasing hormone treatment induces follicular growth and ovulation in seasonally anestrous mares.

A study was conducted to evaluate the effectiveness of gonadotropin-releasing hormone (GnRH) pulse infusion to stimulate follicular development and induce ovulation in seasonally anestrous standardbred mares. Seventeen mares were selected for use in this experiment, on the basis of a previous normal reproductive history, and were housed under a photoperiod of 8L:16D beginning one week prior to the start of the experiment (second week in January). Mares were infused with 20 micrograms (n = 7) or 2 micrograms (n = 6) GnRH/h, or were subjected to photoperiod treatment only (controls, n = 4). Serum concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and progesterone did not vary, and neither significant follicular development nor ovulation was observed in any control mare throughout the experimental period (greater than 60 days). By contrast, both groups of GnRH-treated mares showed elevated serum concentrations of LH and FSH within one day after the start of infusion. Mares infused with 20 micrograms GnRH/h had at least one follicle greater than or equal to 25 mm in 7.4 +/- 1.3 (mean +/- SEM) days following the start of infusion, and ovulated in 12.0 +/- 0.7 days. In the 2-microgram-GnRH/h treatment group, a 25-mm follicle was detected in 5.7 +/- 0.7 days, and ovulation occurred after 10.0 +/- 0.3 days of infusion. Ovulation in every instance was followed by a functional luteal phase, as indicated by the profiles of progesterone secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Anestrus↗

Pregnancy rates for embryos transferred from early postpartum beef cows into recipients with normal estrous cycles.

Survival rate of embryos from first ovulations of postpartum cows with SHORT (6.9 +/- 0.3 days; n = 35) or NORMAL (17.1 +/- 0.3 days; n = 42) luteal phases and quality of the embryos on Day 6 were compared. At 19 to 23 days postpartum, cows were allotted to receive a norgestomet implant for 9 days (normal luteal phase) or to serve as untreated controls (short luteal phase). Calves were weaned 7 days after initiation of treatment to induce behavioral estrus in cows for mating. In 25 cows, growth of the ovulatory follicle was monitored by ultrasonography. On Day 6 after estrus, embryos were recovered nonsurgically, and live embryos were transferred into recipient cows exhibiting normal estrous cycles. The medium used to flush the embryos from the uterus of each donor cow was assayed for prostaglandin F2 alpha (PGF2 alpha). Days from calf removal to estrus and size of ovulatory follicles at ovulation (4.1 +/- 0.3 days and 16.7 +/- 0.7 mm, respectively) did not differ between NORMAL and SHORT cows. Interval from detection of the ovulatory follicle to ovulation was longer in NORMAL (10 +/- 0.7 days) than in SHORT cows (8 +/- 0.6 days; p < 0.05). Rates of recovery of an embryo or ovum (64%), rates of fertilization (65%), and quality or stage of development of Day 6 embryos did not differ between SHORT and NORMAL cows. Overall pregnancy rate from recovered oocytes was 13% for SHORT and 32% for NORMAL cows (p = 0.06); survival of fertilized oocytes was 23% for SHORT and 47% for NORMAL cows (p = 0.08).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Immunoreactive beta-endorphin in human ovaries.

Beta-endorphin (beta-EP) immunostainable cells were demonstrated in human ovarian tissue using a non-cross-reacting anti-beta-EP serum and the avidin-biotin-peroxidase detection technique. In ovaries from ovulating and premenopausal women, beta-EP immunoreactivity was localized in the luteinized cells of theca interna of maturing follicles with almost negligible staining in granulosa cells; cells of primary follicles did not stain. In corpora lutea, luteinized cells in both theca interna and granulosa, layers were equally positive. In postmenopausal ovaries, staining was detectable only in scattered luteinized stromal cells. This is the first report on the presence of immunoreactive beta-EP in human ovaries, in which beta-EP seems to be produced by the same sex cord cells engaged in active steroidogenesis and may be under gonadotropin central regulation. The significance of this finding is discussed.

Endorphins↗

Effect of gonadoreline and naloxone on induction of ovulation and plasma LH in rabbit.

To study the effect of an opiate antagonist receptor (naloxone) on LH secretion, two groups of seven does were randomly allocated to GnRH (gonadoreline, 20 microgram/animal), and GnRH + naloxone (1.8 mg/kg b.w.) treatment, respectively and another group of eight animals was treated with naloxone alone. Blood samples were obtained immediately before treatment (0 minutes) and at 60, 120 and 180 minutes after injection. Plasma levels of LH were determined by a direct, simple and precise sandwich enzyme immunoassay (EIA), adapted for measuring plasma LH in rabbit. Luteinizing hormone reached maximum values 60 minutes after administration of GnRH or GnRH + naloxone (p < 0.05) and all the females of these groups ovulated. Only 25% of naloxone treated rabbit does ovulated and these animals presented a high LH plasma level at 60 minutes post-treatment (p < 0.05). In non ovulated animals no variations in plasma LH concentrations were observed at 60 minutes, in relation to blood samples obtained at 0, 120 and 180 minutes. Results show a limited effect of this naloxone dose on the anterior pituitary of non lactating adult rabbi does in relation to induced ovulation. No synergic effect was detected when naloxone was administered in addition to gonadoreline.

Animals↗