Luteal-phase defect induced by deprivation of FSH at a specific period of the follicular phase prevents pregnancy in the bonnet monkey (Macaca radiata)

Female bonnet monkeys were injected i.v. with 25 µl antiserum to FSH on Days 5, 6 or 7 of the cycle: the length of the luteal phase was shortened but there was no alteration in cycle length. Proven fertile females (N = 6) were caged throughout the period of the experiment (6 cycles) with proven fertile males and treated with 25 µl FSH antiserum on Day 7 of each of 3 successive cycles. Out of 18 cycle exposures during the treatment phase, 17 were ovulatory, but no pregnancies occurred. In the post-treatment phase, 5 monkeys became pregnant within 3 cycle exposures. These results show that it is possible to render female monkeys infertile by creating luteal insufficiency and this can be achieved repeatedly in a reproducible manner by depriving the cyclic females of FSH support on Day 7 of consecutive cycles.

Effects of follicular versus luteal phase-based strength training in young women

Hormonal variations during the menstrual cycle (MC) may influence trainability of strength. We investigated the effects of a follicular phase-based strength training (FT) on muscle strength, muscle volume and microscopic parameters, comparing it to a luteal phase-based training (LT). Eumenorrheic women without oral contraception (OC) (N = 20, age: 25.9 ± 4.5 yr, height: 164.2 ± 5.5 cm, weight: 60.6 ± 7.8 kg) completed strength training on a leg press for three MC, and 9 of them participated in muscle biopsies. One leg had eight training sessions in the follicular phases (FP) and only two sessions in the luteal phases (LP) for follicular phase-based training (FT), while the other leg had eight training sessions in LP and only two sessions in FP for luteal phase-based training (LT). Estradiol (E2), progesterone (P4), total testosterone (T), free testosterone (free T) and DHEA-s were analysed once during FP (around day 11) and once during LP (around day 25). Maximum isometric force (Fmax), muscle diameter (Mdm), muscle fibre composition (No), fibre diameter (Fdm) and cell nuclei-to-fibre ratio (N/F) were analysed before and after the training intervention. T and free T were higher in FP compared to LP prior to the training intervention (P < 0.05). The increase in Fmax after FT was higher compared to LT (P <0.05). FT also showed a higher increase in Mdm than LT (P < 0.05). Moreover, we found significant increases in Fdm of fibre type ΙΙ and in N/F only after FT; however, there was no significant difference from LT. With regard to change in fibre composition, no differences were observed between FT and LT. FT showed a higher gain in muscle strength and muscle diameter than LT. As a result, we recommend that eumenorrheic females without OC should base the periodization of their strength training on their individual MC.

In vivo and in vitro studies on the differential role of luteinizing hormone and follicle-stimulating hormone in regulating follicular function in the bonnet monkey (Macaca radiata) using specific gonadotropin antibodies

Although a distinct need for FSH in the regulation of follicular maturation in the primate is well recognized, it is not clear how FSH controls the functionality of different cellular compartments of the follicle. It is also not evident whether there is a requirement for LH in follicular maturation in the primate. In the first part of the present study, female bonnet monkeys were administered a well-characterized ovine (o) LH antiserum to neutralize endogenous monkey LH for different periods during the follicular phase, and the effect on the overall follicular maturation process was assessed by analyzing serum estrogen (E) and progesterone (P) profiles. Neither continuous LH deprivation from Day 8 of the cycle nor deprivation of LH on any one day between Days 6 and 10 had a significant effect on serum E and P profiles and the follicular maturation process. The period for which the antiserum was effective was dependent upon the dose injected; 1 ml of the antiserum given on Day 8 blocked ovulation but not follicular maturation. To assess the effect of deprivation of LH/FSH at the cellular level, animals were deprived in vivo of LH (on Days 8 and 9 of the cycle) or of FSH (on Day 6 of the cycle) by injection of highly characterized hCG and ovine (o) FSH antisera, respectively; the in vitro responsiveness of granulosa and thecal cells isolated on Day 10 from these animals was then determined.(ABSTRACT TRUNCATED AT 250 WORDS)

Is there a role for estrogen in follicular maturation in the primate

The present study focusses attention on the effects of blocking estrogen synthesis, during follicular phase, on follicular maturation in the adult female bonnet monkey (Macaca radiata). Administration of cycling females (n = 4) with an aromatase inhibitor CGS 16949A (AI) by Alzet mini-pump (2.5 mg/day) from day 3 of cycle resulted in significant reduction in basal (by 53%) and surge levels of estrogen (by 70%) but this had no effect on follicular maturation, ovulation and luteal function as assessed by serum hormone profiles as well as laparotomy. This lack of need for estrogen for completion of follicular maturation process was confirmed by administering cycling monkeys hFSH (25 IU/day) from day 3 till day 8 of the cycle along with (5 mg AI/day) or without Al (n = 3/group). Administration of Al resulted in suppression of FSH induced increase in serum estrogen (by 100%) and elevation in circulating androstenedione. Aromatase inhibitor treatment had no effect on either the number of follicles developed or their size relative to control. Testing the ability of both granulosa and thecal cells, removed on day 9 of treatment cycle, to respond to gonadotropins in vitro showed no change indicating that cellular development and maturation of follicular cells had occurred normally. It is concluded that follicular maturation in the primate can occur even when increase in estrogen synthesis is blocked.

Ovarian stimulation to cryopreserve fertilized oocytes in cancer patients can be started in the luteal phase

OBJECTIVE: To analyze if oocytes can be obtained in all patients before cancer treatment within 2 weeks by initiating ovarian stimulation during the follicular or luteal phase. DESIGN: Prospective controlled multicenter trial. SETTING: Four university-based centers. PATIENT(S): Forty cancer patients before chemotherapy. INTERVENTION(S): Twenty-eight patients were stimulated with gonadotropins in the follicular phase (group I). In 12 patients (group II), ovarian stimulation was initiated in the luteal phase, and these received GnRH antagonists and recombinant FSH. In 14 patients, 143 oocytes were further processed for fertilization by intracytoplasmic sperm injection (ICSI). MAIN OUTCOME MEASURE(S): Number of oocytes aspirated after ovarian stimulation, cumulative FSH/hMG dosage, viability and maturity of oocytes, and fertilization rate by ICSI. RESULT(S): Patients in group I (age 27.6 +/- 4.9 yrs) were stimulated on average for 10.6 days, and patients in group II (age 31.2 +/- 5.7 yrs) for 11.4 days. Total amount of FSH was on average 2,255 IU (I) and 2,720 IU (II) per patient. Average and median numbers of aspirated oocytes were, respectively, 13.1 and 11.5 (I) versus 10.0 and 8.5 (II); 83.7% (I) and 80.4% (II) of the oocytes were mature and viable and could be treated by ICSI. Fertilization rate was 61.0% (I) versus 75.6% (II). CONCLUSION(S): This pilot study suggests that oocytes can be obtained before cancer treatment efficiently irrespective of the phase of the menstrual cycle.

Genetic association and gene expression studies suggest that genetic variants in the SYNE1 and TNF genes are related to menstrual migraine

BACKGROUND: Menstrual migraine (MM) encompasses pure menstrual migraine (PMM) and menstrually-related migraine (MRM). This study was aimed at investigating genetic variants that are potentially related to MM, specifically undertaking genotyping and mRNA expression analysis of the ESR1, PGR, SYNE1 and TNF genes in MM cases and non-migraine controls. METHODS: A total of 37 variants distributed across 14 genes were genotyped in 437 DNA samples (282 cases and 155 controls). In addition levels of gene expression were determined in 74 cDNA samples (41 cases and 33 controls). Association and correlation analysis were performed using Plink and RStudio. RESULTS: SNPs rs3093664 and rs9371601 in TNF and SYNE1 genes respectively, were significantly associated with migraine in the MM population (p = 0.008; p = 0.009 respectively). Analysis of qPCR results found no significant difference in levels of gene expression between cases and controls. However, we found a significant correlation between the expression of ESR1 and SYNE1, ESR1 and PGR and TNF and SYNE1 in samples taken during the follicular phase of the menstrual cycle. CONCLUSIONS: Our results show that SNPs rs9371601 and rs3093664 in the SYNE1 and TNF genes respectively, are associated with MM. The present study also provides strong evidence to support the correlation of ESR1, PGR, SYNE1 and TNF gene expression in MM.

Dynamics of Circulating Concentrations of Gonadotropins and Ovarian Hormones Throughout the Menstrual Cycle in the Bonnet Monkey: Role of Inhibin A in the Regulation of Follicle-Stimulating Hormone Secretion

In higher primates, increased circulating follicle-stimulating hormone (FSH) levels seen during late menstrual cycle and during menstruation has been suggested to be necessary for initiation of follicular growth, recruitment of follicles and eventually culminating in ovulation of a single follicle. With a view to establish the dynamics of circulating FSH secretion with that of inhibin A (INH A) and progesterone (P-4)secretions during the menstrual cycle, blood was collected daily from bonnet monkeys beginning day 1 of the menstrual cycle up to 35 days. Serum INH A levels were low during early follicular phase, increased significantly coinciding with the mid cycle luteinizing hormone (LH) surge to reach maximal levels during the mid luteal phase before declining at the late luteal phase, essentially paralleling the pattern Of P-4 secretion seen throughout the luteal phase. Circulating FSH levels were low during early and mid luteal phases, but progressively increased during the late luteal phase and remained high for few days after the onset of menses. In another experiment, lutectomy performed during the mid luteal phase resulted in significant decrease in INH A concentration within 2 hr (58.3 +/- 2 vs. 27.3 +/- 3 pg/mL), and a 2- to 3-fold rise in circulating FSH levels by 24 hr (0.20 +/- 0.02 vs. 0.53 +/- 0.14 ng/mL) that remained high until 48 hr postlutectomy. Systemic administration of Cetrorelix (150 mu g/kg body weight), a gonadotropin releasing hormone receptor antagonist, at mid luteal phase in monkeys led to suppression of serum INH A and P-4 concentrations 24 hr post treatment, but circulating FSH levels did not change. Administration of exogenous LH, but not FSH, significantly increased INH A concentration. The results taken together suggest a tight coupling between LH and INH A secretion and that INH A is largely responsible for maintenance of low FSH concentration seen during the luteal phase. Am. J. Primatol. 71:817-824, 2009.

Use of a specific aromatase inhibitor for determining whether there is a role for oestrogen in follicle/oocyte maturation, ovulation and preimplantation embryo development

The specific role of oestrogen in follicular maturation, ovulation and early embryonic development was investigated using Fadrozole (CGS 16949A), a non-steroidal aromatase inhibitor, to block oestrogen synthesis specifically and effectively in experimental animals. Induced and normal cyclical follicular maturation as well as normal and hCG/LH-induced ovulation were relatively unaffected by significantly depleting oestrogen in all animals (hamsters, rabbits, monkeys) studied other than rats. Fadrozole treatment significantly reduced the number of healthy antral follicles produced and the ovulatory response to exogenous hCG of immature rats primed with pregnant mares' serum gonadotrophin. The effect was specific, in that exogenously administered oestrogen reversed the blockade. Depletion of oestrogen, starting early in pro-oestrus in hamsters, had no effect on ovulation, oocyte maturation and fertilization, as normal implantation sites were seen on day 6 after coitus. In rabbits, oestrogen depletion during the periovulatory phase affected oviductal morphology and function. Although fertilization was not impaired, early embryo development did not appear to be normal. In monkeys, oestrogen depletion during the follicular phase did not lead to a block of follicular maturation or ovulation but resulted in a significant reduction in secretion of cervical mucus. Administration of either Fadrozole or Tamoxifen during the early luteal phase in cyclic monkeys that were allowed to mate prevented implantation and this appears to be due to impaired fertilization or faulty embryo development. These results suggest that, although there is a clear requirement for oestrogen to support the reproductive cycle in the female, the need for oestrogen in regulating specific events is species dependent.

Dung as a potential medium for inter-sexual chemical signaling in Asian elephants (Elephas maximus)

Chemical signaling is a prominent mode of male-female communication among elephants, especially during their sexually active periods. Studies on the Asian elephant in zoos have shown the significance of a urinary pheromone (Z7-12:Ac) in conveying the reproductive status of a female toward the opposite sex. We investigated the additional possibility of an inter-sexual chemical signal being conveyed through dung. Sixteen semi-captive adult male elephants were presented with dung samples of three female elephants in different reproductive phases. Each male was tested in 3 separate trials, within an interval of 1-3 days. The trials followed a double-blind pattern as the male and female elephants used in the trials were strangers, and the observer was not aware of the reproductive status of females during the period of bioassays. Males responded preferentially (P < 0.005), in terms of higher frequency of sniff, check and place behavior toward the dung of females close to pre-ovulatory period (follicular-phase) as compared to those in post-ovulatory period (luteal-phase). The response toward the follicular phase samples declined over repeated trials though was still significantly higher than the corresponding response toward the non-ovulatory phase in each of the trials performed. This is the first study to show that male Asian elephants were able to distinguish the reproductive phase of the female by possibly detecting a pre-ovulatory pheromone released in dung. (C) 2012 Elsevier B.V. All rights reserved.

Studies on reactivation of regressing bonnet monkey corpus luteum on day 1 of menses: A pilot study

Studies on functional characteristics of the regressing primate corpus luteum (CL) to luteotrophic stimulus on day 1 of the non-fertile menstrual cycle are scarce. Recombinant human luteinizing hormone (rhLH) (20 IU/Kg BW; n = 10) or human chorionic gonadotropin (hCG) (180 IU; n = 6) were administered intravenously to female bonnet monkeys on day 1 of menses. Exogenous treatment of rhLH or hCG caused a significant increase in circulating progesterone (P4) levels 2-4 hours post treatment (P < 0.05). Lutectomy prior to onset of menses confirmed that CL is the site of the increased P4 concentrations. Increased levels of phosphorylated P44/42 MAPK, MKK3/6 activation and concomitant histological changes were observed within 4 hours in CL of monkeys receiving hCG treatment. The results from this study demonstrate the acute progesterone synthesizing capacity of regressing monkey CL after LH or hCG challenge. This has potential implications for interpreting the steroidogenic response after gonadotropin stimulation tests in the early follicular phase of the normal ovulatory and anovulatory women undergoing controlled ovarian stimulation protocols as part of assisted reproductive technology (ART) and in women with polycystic ovarian syndrome.

Menstrual status and thermoregulatory responses of active adolescents during exercise in a cold environment

This study examined the interactions between the reproductive status and the thermoregulatory responses during exercise in the cold in girls involved in competitive sports. Four girls with established menstrual cycles comprised the eumenorrheic menarcheal group (EM) and 5 non-menstruating girls comprised the pre-menarcheal group (PM). During the first visit maximal oxygen consumption, height, weight and percent body fat (%BF) were measured. The second visit involved: a determination of metabolic rate in thermoneutrality (21°C) involving 10-min rest and 20-min cycling (30% of VCL max), and a cold stress test (5°C, 40% humidity, <0.3 m/s air velocity) involving 20-min rest and 40-min cycling (30% of VCL max.). Subjects in the EM group were tested twice in the chamber during the follicular and luteal phases. Pre-menarcheal subjects were found to have significantly (p<0.05) lower core temperatures during the final stages of cold exposure. Overall, body fat was not significantly correlated with core temperature in the cold, however there was a significant surface-to-mass ratio difference between the groups. While in the follicular phase, EM girls had a higher core temperature during cold exposure. Therefore, reproductive hormonal status seems to be an important factor in terms of cold tolerance in females during adolescence.

Effects of dietary restraint and oral contraceptives on bone strength and bone turnover in young women

ABSTRACT Introduction The purpose of this study was to assess specific osteoporosis-related health behaviours and physiological outcomes including daily calcium intake, physical activity levels, bone strength, as assessed by quantitative ultrasound, and bone turnover among women between the ages of 18 and 25. Respective differences on relevant study variables, based on dietary restraint and oral contraceptive use were also examined. Methods One hundred women (20.6 ± 0.2 years of age) volunteered to participate in the study. Informed written consent was obtained by all subjects prior to participation. The study and all related procedures were approved by the Brock University Research Ethics Board. Body mass, height, relative body fat, as well as chest, waist and hip circumferences were measured using standard procedures. The 10-item restrained eating subscale of the Dutch Eating Behaviour Questionnaire (DEBQ) was used to assess dietary restraint (van Strien et al., 1986). Daily calcium intake was assessed by the Rapid Assessment Method (RAM) (Hertzler & Frary 1994). Weekly physical activity was documented by the 4-item Godin Leisure-Time Exercise Questionnaire (Godin & Shephard 1985). Bone strength was determined from the speed of sound (SOS) as measured by QUS (Sunlight 7000S). SOS measurements (m/s) were taken of the dominant and non-dominant sides of the distal one third of the radius and the mid-shaft of the tibia. Resting blood samples were collected from all subjects between 9am and 12pm, in order to evaluate the impact of lifestyle factors on biochemical markers of bone turnover. Blood was collected during the early follicular phase of the menstrual cycle (approximately days 1-5) for all subjects. Samples were centrifliged and the serum or plasma was aliquoted into separate tubes and stored at -80°C until analysis. The bone formation markers measured were Osteocalcin (OC), bone specific alkaline phosphatase (BAP) and 25-OH vitamin D. The bone resorption markers measured were the carboxy (CTx) and amino (NTx) terminal telopeptides of type-I collagen crosslinks. All markers were assessed by ELISA. Subjects were divided into high (HDR) and low dietary restrainers (LDR) based on the median DEBQ score, and also into users (BC) and non-users (nBC) of oral contraceptives. A series of multiple one way ANOVA's were then conducted to identify differences between each set of groups for all relevant variables. A two-way ANOVA analysis was used to explore significant interactions between dietary restraint and use of oral contraceptives while a univariate follow-up analysis was also performed when appropriate. Pearson Product Moment Correlations were used to determine relationships among study variables. Results HDR had significantly higher BMI, %BF and circumference measures but lower daily calcium intake than LDR. There were no significant differences in physical activity levels between HDR and LDR. No significant differences were found between BC and nBC in body composition, calcium intake and physical activity. HDR had significantly lower tibial SOS scores than LDR in both the dominant and non-dominant sites. The post-hoc analysis showed that within the non-birth control group, the HDR had significantly lower tibial SOS scores of bone strength when compared to the LDR but Aere were no significant differences found between the two dietary restraint groups for those currently on birth control. HDR had significantly lower levels of OC than LDR and the BC group had lower levels of BAP than the nBC group. Consistently, the follow-up analysis revealed that within those not on birth control, subjects who were classified as HDR had significantly (f*<0.05) lower levels of OC when compared with LDR but no significant differences were observed in bone turnover between the two dietary restraint groups for those currently on birth control. Physical activity was not correlated with SOS scores and bone turnover markers possibly due to the low physical activity variability in this group of women. Conclusion This is the first study to examine the effects of dietary restraint on bone strength and turnover among this population of women. The most important finding of this study was that bone strength and turnover are negatively influenced by dietary restraint independent of relative body fat. In general, the results of the present thesis suggest that dietary restraint, oral contraceptive use, as well as low daily calcium intake and low physical activity levels were widespread behaviours among this population of college-aged women. The young women who were using dietary restraint as a strategy to lose weight, and thus were in the HDR group, despite their higher relative body fat and weight, had lower scores of bone strength and lower levels of markers of bone turnover compared to the low dietary restrainers. Additionally, bone turnover seemed to be negatively affected by oral contraceptives, while bone strength, as assessed by QUS, seemed unaffected by their use in this population of young women. Physical activity (weekly energy expenditure), on the other hand, was not associated with either bone strength or bone tiimover possibly due to the low variability of this variable in this population of young Canadian women.

Superovulation chez la vache laitière : impact de la progestéronémie

Les récentes recherches suggèrent que la superovulation de vaches laitières sous une concentration élevée de progestérone permet de meilleurs résultats que la superovulation sous une basse progestéronémie pendant la première vague folliculaire. Nous émettions donc l’hypothèse qu’une basse progestéronémie pendant la phase lutéale, une problématique connue chez la vache laitière haute productrice, compromet le rendement en embryons suite à la superovulation de la deuxième vague folliculaire. Afin de tester cette hypothèse, 18 vaches laitières ont été superovulées à deux reprises avec deux protocoles distincts, dont un atteignant un niveau lutéal de progestérone (>2.5 ng/mL, le corps jaune comme source de progestérone) et l’autre un niveau sublutéal (<2.5 ng/mL, l’implant intravaginal comme unique source de progestérone). Le nombre d’embryons transférables était similaire entre les protocoles. Curieusement, nous avons obtenu un développement accéléré des follicules dans le protocole sublutéal (p = 0,002), un développement embryonnaire plus avancé (p = 0,01) et une qualité améliorée des embryons (p = 0,02) par rapport au protocole lutéal. Ces résultats suggèrent que des facteurs autres qu’une basse progestéronémie peuvent affecter le rendement en embryons. Une pulsatilité augmentée de la LH grâce à une basse progestéronémie pendant la deuxième vague folliculaire pourrait être responsable du développement folliculaire accru ainsi que du développement et de la qualité augmentés des embryons. Ces résultats indiquent que des niveaux sublutéaux de progestérone pendant la phase lutéale ne compromettent pas le résultat d’un traitement de superovulation mais, au contraire, peuvent améliorer certains aspects du rendement en embryons.

Expressão gênica de bcl2, receptor de estradiol e receptores de progesterona A e B em endométrio eutópico e ectópico de pacientes inférteis sem e com endometriose

A endometriose caracteriza-se pela presença de tecido endometriótico – glândulas e estroma – fora da cavidade uterina. A doença acomete superfícies peritoniais da pélvis, ovários e septo rectovaginal, sendo mais comum a endometriose pélvica. Embora os dados sobre a prevalência exata de endometriose sejam incertos, as evidências indicam uma prevalência em torno de 5 a 10% das mulheres em idade reprodutiva. Dor, dismenorréia, dispareunia e infertilidade são sintomas freqüentes da doença. Além de apresentar dependência aos hormônios sexuais, a endometriose está associada a um conjunto de desordens de diferentes etiologias, sendo considerada uma doença multifatorial de caráter genético e imune. Neste sentido, a carência de respostas imunológicas adequadas, aliada a um provável desequilíbrio entre os processos de proliferação e apoptose celular, pode ter um papel importante na instalação e manutenção das lesões endometrióticas. O objetivo deste trabalho foi caracterizar a expressão de genes relacionados aos processos de proliferação e diferenciação celular em implantes endometrióticos e endométrio eutópico de mulheres inférteis sem e com endometriose, através da análise semiquantitativa por RT-PCR. Foram estudadas 34 pacientes consultando por infertilidade e submetidas à laparoscopia diagnóstica. Quinze pacientes (31,7 ± 1,5 anos) apresentavam endometriose e 19 (32,9 ± 0,9 anos) foram consideradas controles inférteis sem endometriose. Os dois grupos apresentaram IMC e SHBG similares. As biópsias de endométrio foram realizadas, em fase folicular, nas pacientes do grupo controle (C) e nas pacientes com endometriose (endométrio eutópico (EUT) e ectópico (ECT)). A presença do mRNA para os receptores de estrogênio e progesterona detectada nos 3 grupos estudados, sustenta a existência de sensibilidade tecidual local aos hormônios sexuais. Nas condições experimentais do presente estudo não foi observada diferença estatística na expressão gênica de bcl2 entre os grupos. A expressão do gene do ER endometrial também foi similar entre os grupos controle, eutópico e ectópico. Contudo, o endométrio ectópico apresentou níveis de mRNA mais elevados para PR-A (0,84  0,02 UA) em relação aos grupos controle e eutópico (0,60  0,04 UA e 0,63  0,04 UA; p < 0,05). A expressão gênica do PR-B não foi estatisticamente diferente entre os grupos do presente trabalho. Porém, houve forte correlação negativa entre a expressão dos genes bcl2 e PR-B nas amostras de endométrio sem e com endometriose (r = - 0,795 e p = 0,018), sugerindo que um papel anti-proliferativo do PR-B possa ser operativo neste modelo de estudo.