Follicle growth and endocrine dynamics in women with spontaneous luteinized unruptured follicles versus ovulation.

STUDY QUESTION: Do growth patterns and endocrine profiles differ between ovulatory follicles (OvFs) and luteinized unruptured follicles (LUFs) in women? SUMMARY ANSWER: Growth rates, diameters and associated endocrine profiles differed between OvFs and LUFs in unstimulated cycles. WHAT IS KNOWN ALREADY: Two-three waves of antral follicles develop during the menstrual cycle in ovulatory women of reproductive age, with the second or third wave terminating in ovulation. In contrast, some women can develop LUFs, where a preovulatory follicle fails to rupture and there is subsequent luteinization of the follicle wall. However, no study has compared OvFs and LUFs in unstimulated cycles. STUDY DESIGN, SIZE, DURATION: This retrospective observational study was conducted in 56 healthy women of reproductive age (range: 19-41 years) and with a history of regular menstrual cycles. PARTICIPANTS/MATERIALS, SETTING, METHODS: Participants who met inclusion criteria were enrolled, as previously reported. Daily transvaginal ultrasonography was performed for one interovulatory interval (IOI) to measure the diameters of all follicles >2 mm. Blood samples were collected every 3 days during the IOI to measure serum concentrations of FSH, LH, estradiol and progesterone. MAIN RESULTS AND THE ROLE OF CHANCE: The interval from emergence to deviation (i.e. follicle selection) was shorter (P < 0.05) for LUFs compared to OvFs. However, the intervals from emergence to maximum diameter and deviation to maximum diameter were longer (P < 0.05) for LUFs compared to OvFs. Follicle deviation in LUFs occurred at a larger diameter (P < 0.05) compared to OvFs, and LUFs grew to larger (P < 0.0001) diameters compared to OvFs. Moreover, LUFs grew faster (P < 0.05) from emergence to deviation and from deviation to maximum diameter, compared to OvFs. LUFs were associated with low (P < 0.05) systemic LH levels at emergence and maximum diameter compared to OvFs. LUFs were also associated with low (P < 0.05) systemic FSH and high (P < 0.05) systemic progesterone at deviation and maximum diameter, respectively. Estradiol was higher (P < 0.05) at deviation and lower (P < 0.05) at maximum diameter for LUFs compared to OvFs. LIMITATIONS, REASONS FOR CAUTION: A 3-day interval of blood sampling for hormonal analyses was conducted, as a more frequent sampling interval was not considered acceptable by the study volunteers. A 3-day sampling interval did not allow characterization of acute changes in hormone production during the IOI. In addition, study visits were less frequent when LUFs persisted long after the expected day of the second ovulation of the IOI. WIDER IMPLICATIONS OF THE FINDINGS: Information about the growth and endocrine dynamics of OvFs and LUFs developing in unstimulated cycles in women may be applied to the early detection of LUF-associated anovulatory infertility and clinical management of women with this condition. STUDY FUNDING/COMPETING INTEREST(S): No external funding sources were used for this study. The authors have no conflicts of interest in publishing this manuscript. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov Identifier: NCT01389141.

Aromatase inhibitors: A new approach for controlling ovarian function in cattle.

Numerous treatments and protocols have been used to control the reproductive cycle in cattle, with varying effectiveness and many involving the administration of steroid hormones. Steroid hormones, such as estradiol, are perceived as having a negative impact on consumer health. This internationally shared opinion has led to a ban on the use of steroid hormones in food producing animals in many countries (i.e., European Union, New Zealand, and Australia). Letrozole, a non-steroidal aromatase inhibitor, inactivates the aromatase enzyme responsible for the synthesis of estrogens by reversibly binding to the "heme" group of the P450 subunit. Letrozole is approved as an adjuvant or first-line treatment for hormone-dependent breast cancer in post-menopausal women, but has been used increasingly for ovulation induction in the treatment of infertility in women. Using the bovine model to determine the effects on ovarian function, letrozole treatment was found to extend the lifespan of the dominant follicle and thereby delay emergence of the next follicle wave and/or ovulation. Letrozole treatment also had a luteotrophic effect; that is, larger CL and/or higher circulating concentrations of progesterone were detected in letrozole-treated heifers. Results of the initial studies in cattle provided the impetus for the development of aromatase inhibitor-based synchronization and fertility treatment in cattle. Biologically active concentrations of letrozole were achieved via intravenous, intramuscular or intravaginal administration, but the intravaginal route of administration is of particular interest because it permits extended and defined treatment periods, is minimally invasive, and reduces animal handling. Recent results revealed that irrespective of the stage of the cycle, a 4-day letrozole-based protocol induced ovulation in a significantly greater proportion of animals and with significantly greater synchrony than the control treatment. Evidence and reasons for the increasing use of programmed breeding and fixed-time artificial insemination are discussed in this review as a background to current development of an innovative aromatase inhibitor-based protocol as a safe and effective method of controlling the estrous cycle and ovulation in cattle.

Effects of a single 20 mg dose of letrozole on ovarian function post dominant follicle selection: an exploratory randomized controlled trial.

BACKGROUND: Our objective was to explore the impact of a single dose of an aromatase inhibitor (letrozole) administered at defined times of the follicular phase or immediately after ovulation on dominant follicle development, luteogenesis and new follicle wave emergence. METHODS: A prospective pilot study using a randomized complete block, controlled, open label design was conducted at an academic clinical research center. Forty-five healthy, female volunteers (25.5 ± 0.9 years, BMI 25.0 ± 0.6 kg/m2) who had not taken hormonal contraceptives for a minimum of 2 months were recruited. A 20 mg dose of Letrozole was administered once orally in each of 3 groups when the dominant follicle reached a diameter of 1) 12 mm, 2) 18 mm, 3) the first day following ovulation (post-ovulation), or 4) treatment was withheld (control). Serial ultrasonography and phlebotomy began on day 4 of the menstrual cycle and continued for 1.5 menstrual cycles. Participants recorded menses and daily events in a life events calendar for the duration of the study. Demographic and single point measurements were compared among groups by ANOVA. Changes in hormone concentrations over time were compared among groups by repeated measures ANOVA. Kruskal-Wallis tests were used for non-normally distributed data. RESULTS: The dominant follicle in all treatment groups ovulated. There were no differences among experimental groups in peak follicle diameter, follicular growth rate, endometrial thickness at ovulation or inter-ovulatory interval. Plasma concentrations of estradiol dropped, while FSH and LH concentrations rose following treatment in all treatment groups. Plasma FSH and LH concentrations were higher in the 18 mm group compared to the 12 mm and post-ovulation groups (P < 0.02). CONCLUSION: Administration of a single 20 mg dose of Letrozole at the times of the menstrual cycle we examined did not induce dominant follicle regression or failure of corpus luteum formation. Letrozole-induced suppression of estradiol synthesis by the dominant follicle was not detrimental to follicle growth or ovulation following follicle selection, likely due to increased circulating concentrations of FSH and LH resulting from a lack of estradiol-induced suppression of the hypothalamic-pituitary-ovarian axis. TRIALS REGISTRATION NUMBER: Clinical trials registration number NCT01046578 .

Synchronization of ovulation in cattle with an aromatase inhibitor-based protocol.

A study was designed to determine the effect of stage of the estrous cycle on the proportion of animals that ovulated and the synchrony of ovulation of heifers treated with an aromatase inhibitor-based protocol. Forty-eight heifers were treated intramuscularly with 500 µg of cloprostenol (PGF) followed by 100 µg of GnRH 24 hours later to serve as control data for comparison of the ovulatory response to a subsequent aromatase inhibitor protocol. Daily ultrasound examinations were done to determine the incidence of and interval to ovulation. At the time of ovulation (Day 0), heifers were assigned randomly to five day-groups (n = 8-11/group) and given an intravaginal device containing 3 g of letrozole for 4 days starting on Day 0, 4, 8, 12, or 16. At the time of device removal, heifers were given PGF followed by GnRH 24 hours later. Ultrasound examinations were done daily from 2 days before device insertion to 9 days after the posttreatment ovulation. The preovulatory follicle diameter after letrozole treatment was larger in the Day 4 group compared to the Day 0 and 16 groups and intermediate in the Day 8 and 12 groups (P < 0.001). Compared to control data, the percentage of heifers that ovulated after letrozole treatment was greater (87.1% vs. 69.4%, respectively; P < 0.05) as was the synchrony of ovulation (residuals: 0.24 ± 0.07 vs. 0.68 ± 0.13; P < 0.01). The day on which letrozole treatment was initiated did not affect the proportion of heifers that ovulated or the interval to ovulation. Plasma estradiol concentrations at the time of removal of the letrozole device in the Day 0 and 4 groups was lower (P < 0.05) than in the corresponding controls. Estradiol concentrations in the Day 8 and 12 groups did not differ from already low concentrations in the respective controls. Corpus luteum diameter profiles and progesterone production were not affected by day-group although reduced luteal lifespan after letrozole treatment was observed and requires further investigation. In summary, a protocol involving a letrozole-impregnated intravaginal device for 4 days, PGF treatment at device removal, and GnRH 24 later resulted in a greater ovulation rate and greater synchrony of ovulation than in heifers not given letrozole. Results suggest that the protocol may be initiated effectively at random stages of the estrous cycle and may provide impetus for further studies to assess the efficacy of a letrozole-based synchronization protocol for fixed-time insemination.

Effect of vehicle and route of administration of letrozole on ovarian function in a bovine model.

The objective of this study was to determine the effects of vehicle and route of administration of letrozole on ovarian function in sexually mature beef heifers. On Day 3 (Day 0=ovulation), heifers were assigned randomly to four treatment groups and given 1mgkg(-1) letrozole intravenously (iv, n=10) or intramuscularly (im, n=10) or given a placebo iv (control iv, n=5) or im (control im, n=5). The interwave interval was longer in heifers treated with letrozole im than in im and iv controls (11.7±0.30 vs 9.5±0.50 and 10±0.43, respectively; P<0.05). Corpus luteum diameter profiles and plasma progesterone concentrations were greater (P<0.03 and P<0.05, respectively) in heifers treated with letrozole im compared with control im. Plasma oestradiol concentrations were lower in both letrozole-treated groups compared with controls (P≤0.03). Plasma LH concentrations tended to be elevated at the time of wave emergence in heifers treated with letrozole im compared with other groups (group-by-day interaction, P=0.06) and plasma FSH concentrations tended to be greater (P<0.09) in heifers treated with letrozole by either route compared with a single control group. We conclude that intramuscular administration of letrozole in oil is a feasible route and vehicle for the development of a letrozole-based treatment protocol for herd synchronisation in cattle.

Ultrasound biomicroscopy: a non-invasive approach for in vivo evaluation of oocytes and small antral follicles in mammals.

The use of ultrasonography has changed our understanding of the ovarian function in live animals. However, most of the studies that have used ultrasonography to image the ovary have provided data only of structures >1mm in diameter. The recent availability of high-resolution ultrasound technology with high-frequency transducers (25-70 MHz), offers the potential to examine the developmental dynamics of small antral follicles and the cumulus-oocyte complex (COC) in vivo. In this review we provide data from a series of studies performed by Veterinary Biomedical Sciences Laboratory describing the advantages and disadvantages, as well as image characteristics, of ultrasound biomicroscopy (UBM) to study ovarian biology in mammals. Data and images of small ovarian structures in rabbits, cattle, mice and humans are shown. The UBM technique allowed visualisation of small antral follicles ranging in size from 300 to 700 µm in all species examined, as well as COC within follicles in rabbits, cattle and humans. Furthermore, UBM permitted clear distinction of the follicular wall from the surrounding ovarian stroma in cattle and humans. At present, the limited depth of penetration of UBM restricts the use of this technique to an experimental setting. In that regard, further studies using UBM will probably result in a greater understanding of the pattern and control of early antral folliculogenesis and oogenesis.

In vivo imaging of cumulus-oocyte-complexes and small ovarian follicles in cattle using ultrasonic biomicroscopy.

The objectives of this study were to: (1) evaluate the feasibility of ultrasonic biomicroscopy (UBM) for imaging ovarian follicles in vivo in cows and heifers, (2) compare the transvaginal to the transrectal approach, (3) compare the number of follicles detected by UBM to conventional ultrasonography (US), and (4) identify cumulus-oocyte-complexes (COC) within follicles by UBM. Mature cows (n=5) and peri-pubertal heifers (n=5) were examined once using conventional B-mode US (Aloka 900) with a 7.5 MHz transrectal, and a 5 MHz transvaginal transducers. A second series of examinations was performed using UBM (Visualsonics Vevo 660) equipped with a 30 MHz transrectal and a 40 MHz transvaginal transducer. A three- to four-fold increase in the number of small follicles (<3 mm) was detected using the transvaginal approach with UBM compared to conventional US in both heifers (32.4 ± 4.24 compared to 7.2 ± 1.4; P<0.0001) and cows (35.0 ± 13.8 compared to 10.7 ± 7.5; P=0.0013). However, fewer follicles (all sizes combined) were detected using the transrectal approach with UBM compared to conventional US in both heifers (8.6 ± 5.6 compared to 17.6 ± 4.9; P=0.02) and cows (5.3 ± 6.1 compared to 20.3 ± 7; P=0.04). In heifers, COC-like structures were identified in 39 of 202 (19.3%) follicles examined. In conclusion, UBM using a transvaginal approach is feasible and may be used for in vivo assessment of early antral follicles as small as 0.4 mm, and COC within follicles.

Development of morphologically dominant follicles is associated with fewer metabolic disturbances in amenorrheic women with polycystic ovary syndrome: a pilot study.

OBJECTIVES: To determine if amenorrheic women with polycystic ovary syndrome (PCOS) demonstrate ultrasonographically detectable changes in follicle population. METHODS: Sixteen women with PCOS reporting the absence of menses for more than 3 months were enrolled in the study. Subjects had a physical examination, fasting blood tests and two transvaginal ultrasound scans spaced 1 month apart. In cases where evidence of a morphologically dominant follicle (≥ 10 mm in diameter) occurred, subsequent ultrasound scans were performed to determine the fate of the dominant follicle. Differences in total follicle population, maximum follicle diameter and clinical, hormonal and metabolic features were determined. RESULTS: Forty-four percent of subjects showed changes in follicle population of 6-10 follicles and 37% showed changes in follicle population of > 10. Maximum follicle diameters ranged between 5.4 and 33.0 mm. Four subjects demonstrated follicle diameters ≥ 10 mm. Of those who developed dominant follicles, two subjects ovulated, one subject developed a persistent anovulatory follicle and the dominant follicle regressed in the remaining subject. Diagnostic criteria for PCOS were similar among women that did or did not develop dominant follicles (menstrual cycle length, P = 0.880; hirsutism score, P = 0.809; free androgen index, P = 0.991; total follicle count, P = 0.199). However, lower glycosylated hemoglobin (P = 0.047) and insulin levels (P = 0.049) and better insulin sensitivity (P = 0.048) were noted in women who attained dominant follicles. CONCLUSION: Amenorrheic women with PCOS demonstrate changes in follicle population that are consistent with active follicle growth and regression despite prolonged periods of anovulation. Morphologic selection occurs in amenorrheic women and attainment of dominant follicles is associated with improved metabolic status.

Evaluation of the ultrasound image attributes of developing ovarian follicles in the four follicular waves of the interovulatory interval in ewes.

Computer-assisted quantitative echotextural analysis was applied to ultrasound images of antral follicles in the follicular waves of an interovulatory interval in sheep. The ewe has three or four waves per cycle. Seven healthy, cyclic Western White Face ewes (Ovis aris) underwent daily, transrectal, ovarian ultrasonography for an interovulatory interval. Follicles in the third wave of the ovulatory interval had a longer static phase than that of those in Waves 1 and 2 (P<0.05). The numeric pixel value for the wall of anovulatory follicles emerging in the third wave of the cycle was significantly higher than that for Waves 1 and 2 at the time of emergence (156.7+/-8.09, 101.6+/-3.72, and 116.5+/-13.93, respectively), and it decreased as follicles in Wave 3 reached maximum follicular diameter (P<0.05). The numeric pixel value of the antrum in the ovulatory follicles decreased as follicular diameter increased to > or =5mm in diameter (P<0.05). The pixel heterogeneity of the follicular antrum in Wave 1 increased from the end of the growth phase to the end of the regression phase for follicles in that wave (P<0.05). The total area for the wall and antrum of the follicles studied were correlated with follicular diameter in all follicular waves (r=0.938, P<0.01 and r=0.941, P<0.01 for the wall and antrum, respectively). Changes in image attributes of the follicular wall and antrum indicate potential morphologic and functional differences among antral follicles emerging at different stages of the interovulatory interval in cyclic ewes.

Ultrasonographic characteristics of ovulatory follicles and associated endocrine changes in cyclic ewes treated with medroxyprogesterone acetate (MAP)-releasing intravaginal sponges and equine chorionic gonadotropin (eCG).

The aim of this study was to assess the ultrasonographic characteristics of ovulatory follicles in cyclic Western White Face ewes (December) that had received intravaginal sponges containing medroxyprogesterone acetate (MAP; 60 mg) for 12 days, with or without an injection of 500 IU of equine chorionic gonadotropin (eCG) at sponge removal. We hypothesized that quantitative echotextural attributes of the follicles in ewes treated only with MAP would differ from those in MAP/eCG-treated ewes, reflecting the increased antral follicular growth and secretory function under eCG influence. Digital images of ovulatory follicles obtained at 0 and 24 h after MAP sponge removal and at 24 h before ovulation in the eCG-treated (five ewes, 13 follicles) and control (six ewes, 9 follicles) animals, were subjected to computerized analyses. The mean diameter of ovulatory follicles increased (p < 0.001) 24 h after eCG treatment. The mean pixel intensity and heterogeneity of the follicular antrum (p < 0.001), as well as mean pixel intensity of the follicular wall and perifollicular ovarian stroma (p < 0.05), were greater in eCG-treated animals compared with control ewes 24 h after sponge removal and at 24 h before ovulation. Mean serum concentrations of oestradiol-17beta tended to increase (p = 0.06) 24 h after eCG treatment and the eCG-treated ewes exceeded (p < 0.05) control animals in progesterone concentrations from days 9-15 after ovulation. Our results support the hypothesis that large antral follicles in eCG-treated ewes exhibit distinctive echotextural characteristics. Follicular image attributes in eCG-treated ewes appear to be indicative of the changes in follicular morphology and secretory activity caused by the administration of the exogenous gonadotropin, which has both FSH- and LH-like activities.

Ultrasonographic image attributes of non-ovulatory follicles and follicles with different luteal outcomes in gonadotropin-releasing hormone (GnRH)-treated anestrous ewes.

Ultrasonographic images are composed of multiple square picture elements called pixels. Quantitative changes in numerical pixel values (echotexture) determined by computer-assisted analysis of digital images reflect discrete changes in the microscopic structure and physiological status of ovarian antral follicles. The objective of the present study was to determine and compare the ultrasonographic attributes of non-ovulatory antral follicles that grew to an ostensibly ovulatory diameter (> or =5mm) and follicles with different luteal outcomes in response to gonadotropin-releasing hormone (GnRH) in anestrous Western White Face ewes (n=34). All animals received GnRH injections (250ng i.v. every 2h for 24h) followed by a bolus injection of 125microg of GnRH i.v. Ovarian images obtained by repeated transrectal ultrasonography were digitized and subjected to computerized analyses to determine the changes in follicular size and echotexture of the follicular antrum and wall. At the beginning of GnRH treatment, follicles that formed inadequate corpora lutea following ovulation (ICL; n=22) had higher (P<0.001) pixel intensity of the central and peripheral antrum compared with non-ovulatory follicles (n=40). Pixel intensity of the central follicular antrum was greater (P<0.01) in follicles that formed ICL compared with follicles that formed normal (full-lifespan) CL post-treatment (NCL; n=20) and mean pixel heterogeneity of the follicular wall was greater (P<0.05) in non-ovulatory follicles compared with follicles that gave rise to NCL. At the time of GnRH bolus injection (i.e., induction of a synchronous LH surge), the mean diameter of non-ovulatory follicles was greater (P<0.01) than that of all ovulating follicles, and pixel heterogeneity of the central follicular antrum was lowest (P<0.05) in non-ovulatory follicles. The mean diameter of luteinized unovulated follicles (n=9) tended to be greater (P<0.10) at 2.5 and 3 days after emergence, and pixel intensity of the follicular wall was lower (P<0.05) compared with non-luteinized follicles (n=8) at 1.5 and 2.5 days after emergence (beginning of the growth from approximately 3mm onwards). In conclusion, ovarian antral follicles with different outcomes after GnRH treatment (in seasonally anestrous ewes) had distinctive ultrasonographic characteristics.

Computer assisted image analyses of corpora lutea in relation to peripheral concentrations of progesterone: a comparison between breeds of sheep with different ovulation rates.

Transrectal ovarian ultrasonography is a non-invasive technique that permits the real-time serial visualization of ovarian structures. Previous studies have reported correlations between serum progesterone concentrations and physical characteristics of the corpus luteum (CL) in different species. The aim of the present study was to investigate whether or not there were correlations between ultrasound image attributes of the CL and changing progesterone concentrations over time, in prolific and non-prolific ewes. Ultrasonographic images of CL were taken, once daily, from 12 Western White Face ewes and 7 Finn ewes for the duration of one luteal phase. Blood samples were collected daily prior to each scanning session and assayed to determine mean serum concentrations of progesterone. Analysis of ultrasound images was performed using a series of custom-developed computer algorithms optimized for ultrasonography on a computer graphics workstation. Both total luteal area and mean pixel values were correlated with the pattern of serum concentrations of progesterone from days 3 to 15 after ovulation in Western White Face ewes and from days 3 to 14 in Finn ewes. There was no significant correlation between progesterone concentrations and spot pixel heterogeneity for either Western White Face ewes or Finn ewes. We concluded that pixel heterogeneity is a poor indicator of progesterone secretory ability of the CL when compared to mean pixel values. However, luteal area and mean spot pixel values are better but not strong indicators of the functional status of the CL in cyclic ewes.

Regression and resurgence of the CL following PGF2alpha treatment 3 days after ovulation in mares.

The present study was designed to characterize and compare the physiology and ultrasonographic morphology of the corpus luteum (CL) during regression and resurgence following a single dose of native prostaglandin F2alpha (PGF) given 3 days after ovulation, with a more conventional treatment given 10 days after ovulation. On the day of pre-treatment ovulation (Day 0), horse mares were randomly assigned to receive PGF (Lutalyse; 10 mg/mare, i.m.) on Day 3 (17 mares) or Day 10 (17 mares). Beginning on either Days 3 or 10, follicle and CL data and blood samples were collected daily until post-treatment ovulation. Functional and structural regression of the CL in response to PGF treatment were similar in both the Day 3 and 10 groups, as indicated by an abrupt decrease in circulating concentrations of progesterone, decrease in luteal gland diameter and increase in luteal tissue echogenicity. As a result, the mean +/- S.E.M. interovulatory interval was shorter (P < 0.0001) in the Day 3 group (13.2 +/- 0.9 days) than in the Day 10 group (19.2 +/- 0.7 days). Within the Day 3 group, functional resurgence of the CL was detected in 75% of the mares (12 of 16) beginning 3 days after PGF treatment, as indicated by transient major (6 mares) and minor (6 mares) increases (P < 0.05 and < 0.1, respectively) in progesterone. Correspondingly, mean length of the interovulatory interval was longer (P < 0.03) in mares with major resurgence (15.8 +/- 1.6 days) than in mares with minor (11.2 +/- 1.2 days) and no resurgences (13.5 +/- 0.3 days) in progesterone. Structural resurgence of the CL in the Day 3 group and functional and structural resurgence in the Day 10 group were not detected. In conclusion, PGF treatment 3 days after ovulation resulted in structural and functional regression of the CL and hastened the interval to the next ovulation, despite post-treatment resurgences in progesterone.

Systemic concentrations of hormones during the development of follicular waves in mares and women: a comparative study.

Changes in systemic concentrations of FSH, LH, oestradiol and progesterone during the ovulatory follicular wave were compared between 30 mares and 30 women. Based on a previous study, the emergence of the future ovulatory follicle was defined as occurring at 13.0 mm in mares and 6.0 mm in women, and deviation in diameter between the two largest follicles was expected to begin at 22.7 mm in mares and 10.3 mm in women. Mean FSH concentrations were high in mares during the luteal phase, resulting from statistically identified FSH surges occurring in individuals on different days and in different numbers (mean, 1.5 +/- 0.2 surges/mare); the internadir interval was 3.9 +/- 0.3 days. In contrast, mean FSH in women was low during the luteal phase and increased to a prolonged elevation during the follicular phase. The prolonged elevation was apparent in each individual (internadir interval, 15.2 +/- 0.4 days). Changes in LH or oestradiol concentrations encompassing deviation were not detected in mares, but both hormones increased slightly but significantly between emergence and deviation in women. The hypothesis that a greater number of growing follicles causes a greater predeviation decrease in FSH was supported for mares (r, -0.39; P< 0.04), but a similar negative correlation (r, -0.36) was not significant in women. The hypothesis that the increase in oestradiol during the luteal phase in women was at least partly attributable to luteal-phase anovulatory follicular waves was not supported. Normalization of FSH concentrations to the day of emergence showed maximum value on the day of emergence with a significant increase and decrease on each side of emergence in both species. The day of expected deviation occurred 3 days after emergence during the decline in FSH in both species. These results indicated that the previously reported striking similarities in emergence and deviation between mares and women during the ovulatory follicular wave are associated with species similarities in the temporal relationships between follicle events and FSH concentration changes. Thus, mares may be useful research models for studying the role and mechanism of the action of FSH in emergence and deviation during the ovulatory follicular wave in women.

Form and function of the corpus luteum during the human menstrual cycle.

OBJECTIVE: To characterize the growth and regression of the corpus luteum (CL) during an interovulatory interval (IOI) using serial transvaginal ultrasonography. METHODS: Fifty healthy women of reproductive age with a history of regular menstrual cycles underwent daily transvaginal ultrasonography for one IOI. Measurements of luteal area and luteal numerical pixel value (NPV) were recorded each day after ovulation until the CL could no longer be detected. Blood was drawn every third day during the IOI to measure serum concentrations of progesterone and estradiol-17beta. RESULTS: Corpora lutea were of two morphological types: those with a central fluid-filled cavity (CFFC) (78%) and those without (22%). Eighty-eight percent of women exhibited a CL containing a CFFC 2 days after ovulation, followed by 34% 13 days after ovulation and 2% 27 days after ovulation. Luteal area, progesterone concentration and estradiol concentration increased for approximately the first 6 days following ovulation followed by a subsequent decline. Luteal NPV decreased from days 1 to 11 and increased during days 11-16. Changes in luteal area, NPV, progesterone and estradiol concentrations did not differ in women with two versus three waves of follicular development. CONCLUSIONS: Peak luteal function, as determined by maximum luteal area, progesterone concentration and estradiol concentration, is observed 6 days following ovulation. Luteal NPV is reflective of morphological and endocrinological changes in the CL. The development of a CFFC during luteinization is a normal physiological phenomenon. The CL can be detected, but is not functional, during the follicular phase of the menstrual cycle.

Ovarian follicular development is initiated during the hormone-free interval of oral contraceptive use.

We evaluated ovarian follicular development in women during compliant use of oral contraceptives (OC). Thirty-six healthy women received: [35 microg ethinyl estradiol (21)/180 microg norgestimate (7), 215 microg norgestimate (7), 250 microg norgestimate (7)]; [30 microg ethinyl estradiol (21)/150 microg desogestrel (21)]; or [20 microg ethinyl estradiol (21)/100 microg levonorgestrel (21)] for 3 consecutive 28-day cycles. Transvaginal ultrasonography was performed every third day to monitor follicular development. If a follicle reached > or = 14 mm, ultrasonography was performed daily and blood drawn every other day to determine estradiol-17beta concentrations. Seventeen of 36 women (47%) grew follicles > or = 10 mm. Nine of the 17 women (53%) grew follicles > or = 14 mm, in association with increased serum concentrations of estradiol-17beta. Thirty-seven of 43 follicles > or = 10 mm (86%) emerged during the hormone-free interval (HFI). No ovulations were observed. Our results supported the hypothesis that follicular development to an ostensibly ovulatory diameter occurs during compliant OC use, in association with loss of endocrine suppression during the HFI.

Endometrial development in association with ovarian follicular waves during the menstrual cycle.

OBJECTIVES: Waves of ovarian follicular development during the menstrual cycle have recently been documented in our laboratory. The objective of this study was to test the hypothesis that ultrasonographically detectable changes in the endometrium during the menstrual cycle would differ between women with two vs. three waves of ovarian follicular development and among women with different major and minor wave patterns of follicle growth. METHODS: Fifty women of reproductive age (mean age +/- SD, 28.0 +/- 6.9 years) underwent daily transvaginal ultrasonography for one interovulatory interval (IOI). Ultrasonographic images of the endometrium were obtained each day, and measurements of endometrial area and perimeter (based on the shape of an ellipse, in the transverse plane) and thickness and pattern (in the sagittal plane) were recorded. Endometrial area, perimeter, thickness and pattern were compared between women with two and three waves of follicle development and among women with different minor and major wave patterns of follicular growth during the IOI. RESULTS: Endometrial area, perimeter, thickness and pattern increased earlier during the follicular phase in women with two vs. three waves of follicular development. In women with two follicle waves, endometrial area and perimeter increased earlier in those with major major vs. minor major follicle wave patterns. CONCLUSIONS: Ultrasonographically detectable changes in the endometrium occurred in association with follicle wave dynamics in women. Earlier development of the endometrium during the follicular phase in women with two vs. three follicle waves was attributed to an earlier increase in dominant follicle estradiol production.

Comparative study of the dynamics of follicular waves in mares and women.

Deviation in growth rates of the follicles of the ovulatory wave begins at the end of a common growth phase and is characterized by continued growth of the developing dominant follicle (F1) and regression of the largest subordinate follicle (F2). Follicle diameters during an interovulatory interval were compared between 30 mares and 30 women, using similar methods for collecting and analyzing data. Follicles were tracked and measured daily by ultrasonography. Diameter at follicle emergence (mares, 13 mm; women, 6 mm) and the required minimal attained diameter for assessment of follicles (mares, 17 mm; women, 8 mm) were chosen to simulate the reported ratio between the two species in mean diameter of F1 at the beginning of deviation (mares, 22.5 mm; women, 10.5 mm). F1 emerged before F2 (P < 0.02) in each species, and the interval between emergence of the two follicles was similar (not significantly different) between species. Growth rate for F1 and F2 during the common growth phase was similar within species, and the percentage of diameter increase was similar between species. Proportionality between species in diameter of F1 at deviation (2.2 times larger for mares than for women) and at maximum preovulatory diameter (2.1 times larger) indicated that relative growth of F1 after deviation was similar between species. A predeviation follicle was identified in 33% of mares and 40% of women and was characterized by growth to a diameter similar to F1 at deviation but with regression beginning an average of 1 day before the beginning of deviation. The incidence of a major anovulatory wave preceding the ovulatory wave was not different between species (combined, 25%). Results indicated that mares and women have comparable follicle interrelationships during the ovulatory wave, including 1) emergence of F1 before F2, 2) similar length of intervals between sequential emergence of follicles within a wave, 3) similar percentage growth of follicles during the common growth phase, and 4) similar relative diameter of F1 from the beginning of deviation to ovulation. Similar follicle dynamics between mares and women indicate the mare may be a useful experimental model for study of folliculogenesis in women, with the advantage of larger follicle size.

Luteogenesis in cyclic ewes: echotextural, histological, and functional correlates.

To date, it has not been possible to detect corpus luteum (CL) by ultrasonography, immediately following ovulation, in the ewe. Early CL detection is essential to be able to relate luteal outcome to the developmental pattern of the ovulated follicle and to confirm ovulation. Image analysis of the CL may be useful in providing a noninvasive picture of CL differentiation and function. The present study was designed to use high-resolution ultrasonography to monitor and to correlate the echotextural, histological, and functional attributes of the developing ovine CL from Days 1 to 3 after ovulation. Ten ewes underwent twice-daily transrectal ultrasonography and blood sampling from the day of synchronized estrus. Ewes were ovariectomized at 12-24, 36-48, and 60-72 h after ovulation. Ovaries collected were scanned in a water bath before processing for histology. Ultrasonographic images of CL were analyzed for echotexture. Histological sections were analyzed for the percentage area of the CL occupied by blood clot or luteal tissue. Serum samples were analyzed for progesterone concentration. Numerical pixel value, heterogeneity, and percentage of the CL occupied by blood clot declined (P<0.05) from 12-24 to 60-72 h after ovulation. Luteal area and serum progesterone concentration increased (P<0.05) from 12-24 to 60-72 h. The results indicated that it was possible to visualize developing CL as early as 12-24 h after ovulation in the ewe. Echotexture of the CL was closely associated with its morphological and functional characteristics; image analysis holds promise for noninvasive monitoring of CL differentiation and growth.

Ultrasound image characteristics of ovarian follicles in relation to oocyte competence and follicular status in cattle.

Assessment of the quality of the female gamete has become paramount for in vitro procedures. There is a need to identify reliable indicators of oocyte competence and develop a simple, non-invasive method to assess competence. The aim of this study was to investigate the relationships among ultrasonographic attributes of a follicle, its stage of development and the competence of the oocyte that it contains. We tested the hypotheses that follicular echotexture characteristics are related to: (1) the phase of development of the follicle, (2) the presence of the corpus luteum (CL) and/or the dominant follicle in the ovary, and (3) developmental competence of cumulus oocyte complexes (COC) from the same ovary. Crossbred beef cows (n=143), age 4-14 years, were given a luteolytic dose of dinoprost to cause ovulation. Ultrasound-guided ablation of all follicles > or = 4mm was done 8 days later to induce new follicular wave emergence during a luteal phase. Ultrasonographic images of dominant follicles and the three largest subordinate follicles (n=402 follicles; 84 cows) were acquired on Days 2, 3, 5 or 7 of the follicular wave (Day 0: wave emergence), i.e. growing, early-static, late static, and regressing phases of subordinate follicle development, respectively. From a subset of these animals (n=33), ovaries were collected within 30 min of slaughter and COC from subordinate follicles > or = 3mm underwent in vitro maturation, fertilization and culture to the blastocyst stage.Image analysis revealed differences in echotexture between dominant and subordinate follicles among Days 2-7 of the follicular wave. Images of dominant and subordinate follicles at Day 7 of the wave displayed consistently lower grey-scale values (P<0.05) in the peripheral antrum, follicular wall and perifollicular stroma than all other days. Follicle images displayed a consistent pattern of variation in echotexture among follicular phases. Data did not support the hypothesis of a local effect of the CL or dominant follicle on follicular echotexture. Echotexture values of the perifollicular stroma were lower in ovaries that did not produce embryos compared to ovaries that produced embryos. Our results showed that the changes in follicular image attributes are consistent with changes in follicular status. The sensitivity of the technique is not yet sufficient for use in a diagnostic setting, but results provide rationale for further development of image analysis as a tool for evaluating oocyte competence in situ.