Total Number Of Ovarian Follicles Research Articles

Effect of a GnRH Antagonist on Anti-Müllerian Hormone and Follicular Population in Anestrous Bitches.

The objective of this study was to describe the effect of the third-generation GnRH antagonist, acyline, on ovarian follicular population and serum anti-Müllerian hormone (AMH) concentrations in female dogs. Four late anestrous bitches were administered 330 μg/kg SC acyline every 10 days for 60 days and followed up for 45 days. Blood samples were drawn on days -1, 15, 30, 45, 60, 75, 90 and 105 for AMH determination. Then, the females were ovariectomized and the excised ovaries were gross and histologically evaluated. The total ovarian follicles were counted. None of the female dogs presented estrus during treatment. Only one bitch presented an ovulatory estrus 20 days after treatment. The total number of ovarian follicles in these bitches was 96,200.10 ± 26,125.12, with 84.13%, 11.36%, 7.8% and 0.01% corresponding to primordial, primary, secondary and antral structures, respectively. Pretreatment AMH concentrations were 0.62 ± 0.17 ng/mL. This hormone varied throughout the study period (p < 0.01), diminishing to nadir values during treatment to then rapidly recover after its effect (0.2 ± 0.05 vs. 0.67 ± 0.22 ng/mL; p < 0.01). Acyline rapidly and reversibly prevented the initiation of cycling without affecting follicle count but diminishing serum AMH concentrations.

Feeding level is associated with altered liver transcriptome and follicle selection in hen†.

Genetic selection for particular traits in domestic animals may have altered the optimal feedback regulation among systems regulating appetite, growth, and reproduction. Broiler breeder chickens have been selected for fast and efficient growth and, unless feed restricted, consume excessively resulting in poor reproductive efficiency. We examined the effect of dietary treatment in full-fed and restricted-fed broiler breeder hens on ovarian responses, liver morphology, and transcriptome associated with reproductive function. Although full-fed broiler breeder hens had lower egg production (P < 0.01), the total number of ovarian follicles >8mm (P < 0.01), 6-8mm (P < 0.03), and 3-5mm (P < 0.04) were greater in full-fed hens compared to restricted-fed hens. There was a large amount of lipid accumulation in the liver of full-fed hens and differential gene analysis yielded 120 genes that were differentially expressed >2-fold in response to feeding level (P < 0.01; false discovery rate< 0.05). Elevated T3 may indicate that general metabolism was affected by diet and GHR (P < 0.01) and insulin like growth factor 1 (IGF1) (P < 0.04) mRNA expression were both greater in the liver of full-fed hens as compared to restricted-fed hens. It is likely that selection for increased growth, associated with enhanced activity of the IGF1 system, has altered nutritional coupling of feed intake to follicle development.

Long‐term reproductive evaluation of a bitch neonatally treated with a gonadotropin‐releasing hormone agonist

AbstractIn mammals, high concentrations of gonadotropins are necessary early in life to reach a full‐sized pool of ovarian follicles at the age of puberty. This case report describes the reproductive status of a 28‐month‐old, 9.1 kg, crossbred bitch (Bitch 1) which had been treated neonatally with 18.8 deslorelin acetate subcutaneously for contraceptive purposes. The female had just passed pubertal oestrus when hormones were measured and then spayed. The ovaries were histologically assessed. Two crossbred bitches were used for comparisons. Bitch 1's anti‐Müllerian hormone (AMH) and progesterone serum concentrations were 0.35 and 16.7 ng/ml, respectively. The total number of ovarian follicles and proportions were normal and the number of primary and antral follicles was increased. These findings might suggest an enhanced recruitment of the follicle pool to the growing population which could be related to the relatively low AMH concentrations.

52 Ovarian Follicular Profiling of High- and Low-immunocrit Gilts on Postnatal Day 14

Abstract Colostrum intake by neonatal piglets can be measured using the immunoglobulin immunocrit assay (iCrit). Lactocrine effects occur when maternally derived, milk-borne bioactive factors are transferred to the neonatal circulation with consumption of colostrum during nursing and affect development of somatic tissues, which can have long-term consequences in adulthood. Lactocrine deficiency, indicated by low neonatal iCrit, altered uterine gene expression and reduced fecundity in adult, neonatally lactocrine-deficient gilts. Litter size in pigs is dependent on both ovarian and uterine function. It was hypothesized that lactocrine deficiency affects development of ovarian follicles in gilts. The objective was to determine the number of primordial, primary, and secondary follicles in ovaries of gilts with high (12% ± 0.5; n = 10) or low (1.9% ± 0.4; n = 10) iCrit, determined on postnatal day (PND) 1 after birth. Paired high- and low-iCrit gilts were chosen from the same litters (birth weight; 3.1 ± 0.2 lbs). On PND 14, ovaries were collected and histological sections prepared (3 sections per animal; 30–150 µm apart). Ovarian follicles in each section were staged and the number of follicles in each category were quantified and subjected to ANOVA. Total number of ovarian follicles did not differ with iCrit (P = 0.55; 1,370.6 ± 147.8 follicles per section). The proportion of primordial, primary, and secondary follicles was 89.6 ± 1.15%, 7.7 ± 0.87%, 2.7 ± 0.51%, respectively. The number of primordial (P = 0.55), primary (P = 0.64), and secondary (P = 0.93) follicles did not differ with iCrit. Results indicate that ovarian follicular development of neonatal gilts is not sensitive to immunocrit status. Although lactocrine deficiency did not influence the ovarian follicular profile at PND 14, it remains unknown whether lactocrine programming alters ovarian follicular dynamics in neonatally lactocrine-deficient adults. USDA is an equal opportunity provider and employer.

Factors Affecting Ovarian Activity in Donkeys: The Effect of Season, Age and Body Condition Score (BCS)

During the period of year (representing spring, summer, autumn, and winter) Jennies at Giza Zoo abattoir were checked for body condition score (from 1 to 5), age (1 to 7 years; 8 to 14 years; and &gt;15 years old). After culling, ovaries of 377 Jennies were collected, two ovaries were weighed, and the number and size of ovarian follicles were recorded. Cumulus-Oocyte Complexes (COCs) were collected after the ovarian follicles have been sliced and scraped, and COCs number and quality were determined. Data indicated that right and left ovaries and overall ovarian weight were significantly increased (P&lt;0.05) in summer than the other seasons. The number of small size ovarian follicles was decreased (P&lt;0.05) in spring than other seasons, while oocytes number and quality were not significantly affected by season. Jennies of 1 to 7 years old showed a significant (P&lt;0.05) smaller left ovary and total ovarian weight than the two other groups. Jennies of 8-14 years old) have a significant high (P&lt;0.05) small size and a whole number of ovarian follicles than the other groups. Jennies with BCS-3 possess a significantly (P&lt;0.05) higher total ovarian weight, total number of ovarian follicles and the total number of COCs recovered than BCS-1 or BCS-4. While the BCS-4 group showed the lowest (P&lt;0.05) number of ovarian follicles and produced less (P&lt;0.05) number of COCs than the other groups. Conclusion: In Egyptian Jennies, ovarian activity was affected by season, BCS and age of the animal, while, COCs yield and quality were more affected by BCS.

Light-exposure at night impairs mouse ovary development via cell apoptosis and DNA damage.

The alternation of light and dark rhythm causes a series of physiological, biochemical and metabolic changes in animals, which also alters the growth and development of animals, and feeding, migration, reproduction and other behavioral activities. In recent years, many studies have reported the effects of long-term (more than 6 weeks) illumination on ovarian growth and development. In the present study, we observed the damage, repair and apoptosis of ovarian DNA in a short period of illumination. The results showed that, in short time (less than 2 weeks) illumination conditions, the 24-h light treatment caused the reduction of total ovarian follicle number and down-regulation of circadian clock related genes. Furthermore, the changed levels of serum sex hormones were also detected after 24-h light exposure, of which the concentrations of LH (luteinizing hormone), FSH (follicle-stimulating hormone) and E2 (estradiol) were increased, but the concentration of PROG (progesterone) was decreased. Moreover, 24-h light exposure increased the expression of DNA damage and repair related genes, the number of TUNEL and RAD51 positive cells. These results indicated that 24-h light exposure for 4, 8 and 12 days increased DNA damage and cell apoptosis, thereby affecting the development of ovary.

Involvement of Quebracho tannins in dietalters productive and reproductive efficiency of postpartum buffalo cows.

This study was conducted to investigate the effects of 10 weeks supplementation of Quebracho tannins (QT; 0 [control], 100 [QT100] or 200 g/[cow·d] [QT200]) to 30 multiparous postpartum buffalo cows (10 cows per group) on milk yield and composition, blood metabolites and reproductive performance. Supplementation of QT100 had no significant effect on milk yield, whereas QT200 decreased (P < 0.05) this trait. Compared with the control group, both QT levels decreased (P < 0.05) fat-corrected milk (FCM) yield, but no significant effects were found on percentages of milk fat and protein. Contrariwise, yields of milk fat, lactose and milk protein were decreased (P < 0.05) when QT200 was supplemented. The solids nonfat (SNF) percentage and yield were decreased (P < 0.05) with QT100 supplementation. Moreover, QT tended to numerically reduce total number of ovarian follicles, number of small follicles, peripheral progesterone concentration and conception rate. Supplementation of QT200 numerically increased number of large follicles, mean diameter of large follicle, number and diameters of corpora lutea. The inclusion of QT200 shortened days open (DO) and decreased number of services per conception. Contrariwise, QT did not show significant effects on serum total protein, albumin, globulin, glucose, cholesterol and triglycerides concentrations. Supplementation of QT100 caused an increase (P < 0.05) of serum urea compared with that in control and QT200 groups. Generally, QT decreased (P < 0.05) serum creatinine concentration. Therefore, the supplementation of a commercial QT to early lactating Egyptian buffalo cows displayed negative consequences on their productive and reproductive performances.

Radiation-induced ovarian follicle loss occurs without overt stromal changes.

Radiation damage due to total body irradiation (TBI) or targeted abdominal radiation can deplete ovarian follicles and accelerate reproductive aging. We characterized a mouse model of low-dose TBI to investigate how radiation affects the follicular and stromal compartments of the ovary. A single TBI dose of either 0.1 Gy or 1 Gy (Cesium-137 γ) was delivered to reproductively adult CD1 female mice, and sham-treated mice served as controls. Mice were euthanized either 2 weeks or 5 weeks post exposure, and ovarian tissue was harvested. To assess the ovarian reserve, we classified and counted the number of morphologically normal follicles in ovarian histologic sections for all experimental cohorts using an objective method based on immunohistochemistry for an oocyte-specific protein (MSY2). 0.1 Gy did not affect that total number of ovarian follicles, whereas 1 Gy resulted in a dramatic loss. At two weeks, there was a significant reduction in all preantral follicles, but early antral and antral follicles were still present. By five weeks, there was complete depletion of all follicle classes. We examined stromal quality using histologic stains to visualize ovarian architecture and fibrosis and by immunohistochemistry and quantitative microscopy to assess cell proliferation, cell death and vasculature. There were no differences in the ovarian stroma across cohorts with respect to these markers, indicating that this compartment is more radio-resistant relative to the germ cells. These findings have implications for reproductive health and the field of fertility preservation because the radiation doses we examined mimic scatter doses experienced in typical therapeutic regimens.

Effects of diet and arginine treatment during the luteal phase on ovarian NO/PGC-1α signaling in ewes

The aim of this study was to determine whether arginine (Arg) supplementation of malnourished ewes affects the expression of key NO/PGC-1α signaling pathway genes in the ovary. On Day 6–15 of the estrous cycle, 24 multiparous Hu sheep (BW = 43.56 ± 1.53 kg) were randomly assigned to three groups: control group (CG; n = 6), restriction group (RG; n = 9) and l-arginine group (AG; n = 9), and administered Arg treatment (or vehicle) three times per day. The ewes were slaughtered at the end of treatment, and blood samples and ovaries were collected for analysis. The results of our analyses showed that both short-term feed-restriction and/or supplementation with L-Arg-HCl affected the number of different size follicles observed in the ovary, and the relative day of estrus behavior initiation of ewes. Specifically, the relative day of estrus behavior initiation was significantly advanced in AG compared with that in RG ewes (P < 0.05). Both the number of ≤2 mm-ovarian follicles (P < 0.05) and the total number of ovarian follicles (P < 0.05) were significantly increased in the RG and AG compared with that in the CG ewes. RG ewes exhibited a higher proportion of ≤2 mm (P < 0.05), but a lower proportion of >5 mm follicles than did CG ewes (P < 0.05). The mean number of corpus lutea ≥5 mm was significantly increased in AG as compared to that in either CG or RG ewes. Furthermore, the expression of eNOS, nNOS, iNOS, PDE5A, PDE9A, PRKG2, and PPARGC1A varied significantly among the treatment groups (P < 0.05). GUCY1A3 mRNA levels were significantly increased in RG and AG as compared to those in CG ewes (P < 0.05), whereas conversely, GUCY1B3 mRNA levels were significantly decreased in CG and RG as compared to those in AG ewes (P < 0.05). P53 mRNA levels were found to vary significantly among the three experimental treatment groups (P < 0.05), and similarly, the relative expression levels of P53 were greater in AG and RG than in CG ewes (P < 0.05). The levels of eNOS protein were significantly higher in RG than in either CG or AG ewes (P < 0.05). The relative expression levels of PGC-1α were significantly higher in RG (P < 0.05) and significantly lower in AG ewes (P < 0.05) than in CG ewes. In conclusion, the results of the present study indicate that feed-restriction negatively affects follicular development, and that Arg-supplementation may modulate the expression of key NO/PGC-1α signaling pathway genes in the ovary and thereby accelerate ovulatory processes and the estrous rate. Elucidation of mechanisms underlying these effects of Arg on gene expression in the ewe ovary requires further investigation.

115 ROLE OF THE OVARY AND UTERUS FOR THE PLAINS VISCACHA (LAGOSTOMUS MAXIMUS MAXIMUS, CHINCHILLIDAE) REPRODUCTION

A dogma of mammalian reproduction states that primordial germ cells in females are restricted to the intrauterine phase and only small portions of oocytes are available for ovulation during the adult life. Among the rare exceptions to this rule is the plains viscacha. It polyovulates up to 800 oocytes per cycle, from which 10 to 12 are implanted, but only 1 to 2 conceptuses survive. To better understand the main mechanisms involved in these patterns of super-ovulation, super-implantation, and embryonic loss in the viscacha, we conducted an analysis of the ovary and uterus of pregnant females and their conceptuses. Pregnant females (n = 16) of ~50 and 90 days of gestation (early to mid-gestation) were selected for conceptus recovery. Hemi-ovariohysterectomy was performed following surgical and anaesthetic protocols used for laboratory animals and the conceptuses collected. Female fetuses of Day 50 (n = 2) and 90 (n = 1) of gestation were obtained from Estación de Cría de Animales Silvestres, Buenos Aires, Argentina. The reproductive organs were investigated by means of gross morphology, histology (hematoxilin and eosin), stereology (quantification of the volume of the ovary and number of ovary follicles), immunohistochemistry (PCNA, Oct-4, VEGF, and Caspase-3), and transmission electron microscopy. In the Day 50 fetal samples, the ovaries had an ovoid shape with smooth surface without apparent folds. First steps of subdivision were observed in the ovary of fetus of 90 days. The total volume of all fetal ovaries was of 4.8 mm2 and a coefficient of variation (CV) of 0.32. The ovaries of adult individuals had remarkable invaginations with surface projections and were small, asymmetrical, and dorsoventrally flattened with a mean of 77.6 mm3 (CV = 0.47). Only adult females had differentiation of germ cells. Primordial follicles had a mean of 9.9 × 105 (CV = 0.19), representing 93% of the total number of ovarian follicles. The mean of primary follicles was 3.05 × 104 (CV = 0.36), whereas for secondary follicles it was 2.75 × 104 (CV = 0.50), each representing 3% of all ovarian follicles. The number of antral follicles in several stages of development was 8.64 × 103 (CV = 0.75), representing 1% of the follicles. Primordial follicles expressed pluripotency (Oct-4+) and proliferation (PCNA+) markers, as well as the primary follicles. The cells did not react for Caspase-3 as marker for apoptosis. Variations regarding to the vascularization of the different regions of the uterine horn were observed, which were more intense and efficient near to the cervix. Data showed that a specialised, highly convoluted structure of the ovarian cortex developed in the intrauterine phase as a prerequisite for massive super-ovulation, associated with the inhibition of apoptosis and continued proliferation of germ cells, as well as maintenance of several corpora lutea during the adult life. Thus, a highly complex pattern of polyovulation, polyimplantation, and controlling mechanisms has evolved in the female reproductive system of the viscacha that mainly was associated with the maternal side. After an in-depth analysis of the arterial and venous vascularization of the uterine horns and uteri, we speculate that specializations regarding the vasculature and musculature evolved first and then contributed as a compensatory or controlling mechanism for polyovulation and polyimplantation. In conclusion, polyovulation in the viscacha represents a unique enigma in reproductive biology.

English

Boran is an endangered breed of cattle indigenous to Ethiopia and the relatively poor understanding of its reproductive physiology has impeded efforts to maximize reproductive performance of the breed. This study characterized ovarian follicular dynamics in 9 purebred Boran and 8 Boran&times;Holstein (B&times;H) crossbred cows. Ovaries of all 17 cows were examined once per day for 61 consecutive days (encompassing three periods of estrus) using transrectal ultrasonography. The mean (&plusmn;standard error of mean) inter-ovulatory interval (IOI) was similar (P&gt;0.05) in Boran (19.4 &plusmn; 0.2 days) and B&times;H cows (20.1 &plusmn; 0.4 days). Two (in 79% of estrous cycles) or three (in 21% of cycles) follicular waves per IOI were observed and IOI was shorter (P&lt;0.001) for cows with two follicular waves per estrous cycle (n=27; 19.5 &plusmn; 0.9 days) than for cows with three (n=7; 20.9 &plusmn; 2.1 days). All Boran cows (100%) exhibited two waves of follicular growth per estrous cycle and this was higher (P&lt;0.001) than the proportion observed in B&times;H cows (56%). Mean diameter of the dominant follicle was similar (P&gt;0.10) in both genotypes (15.8 &plusmn; 1.5 mm in Boran and 19.4 &plusmn; 2.9 mm in B&times;H). Boran cows possessed a greater (P&lt;0.001) total number of ovarian follicles than B&times;H cows and both genotypes displayed more (P&lt;0.05) activity on their right than left ovary. Results of our study have provided novel insights into the normal reproductive physiology of the Boran breed. Key words: Boran, estrous cycle length, follicular dynamics, follicular waves, dominant follicle.

Impact of Quebracho Tannins Supplementation on Productive and Reproductive Efficiency of Dairy Cows

This study was conducted to investigate effects of supplementing two doses of quebracho tannins (QT; 100 or 200 g/cow/d; QT100 or QT200) pre and post parturition to thirty Holstein dairy cows on Dry Matter Intake (DMI), milk yield and composition, blood metabolites and reproductive performance for 12 weeks. There were no significant changes in DMI due to QT at transition period of dairy cows. QT supplementation at either level decreased (P 0.05) the total number of ovarian follicles, number of large follicles, diameter of largest follicle, number of small follicles, number and diameter of corpus luteum, and progesterone concentration. The inclusion of QT increased days open and number of services per conception, which consequently decreased conception rate, compared to the control cows. QT100 decreased (P < 0.05) serum total protein, globulin, glucose and triglycerides concentrations as compared with their values in control cows. Thus, the supplementation of commercial QT to dairy cows at their transition period had negative impacts on productive and reproductive performance.

MYBL2 guides autophagy suppressor VDAC2 in the developing ovary to inhibit autophagy through a complex of VDAC2-BECN1-BCL2L1 in mammals

Oogenesis is essential for female gamete production in mammals. The total number of ovarian follicles is determined early in life and production of ovarian oocytes is thought to stop during the lifetime. However, the molecular mechanisms underling oogenesis, particularly autophagy regulation in the ovary, remain largely unknown. Here, we reveal an important MYBL2-VDAC2-BECN1-BCL2L1 pathway linking autophagy suppression in the developing ovary. The transcription factors GATA1 and MYBL2 can bind to and activate the Vdac2 promoter. MYBL2 regulates the spatiotemporal expression of VDAC2 in the developing ovary. Strikingly, in the VDAC2 transgenic pigs (Sus scrofa/Ss), VDAC2 exerts its function by inhibiting autophagy in the ovary. In contrast, Vdac2 knockout promotes autophagy. Moreover, VDAC2-mediated autophagy suppression is dependent on its interactions with both BECN1 and BCL2L1 to stabilize the BECN1 and BCL2L1 complex, suggesting VDAC2 as an autophagy suppressor in the pathway. Our findings provide a functional connection among the VDAC2, MYBL2, the BECN1-BCL2L1 pathway and autophagy suppression in the developing ovary, which is implicated in improving female fecundity.

Influence of puberty and antral follicle count on calving day in crossbred beef heifers

Lifetime productivity and longevity are greater in beef cows that give birth early in their first calving season. The ability of heifers to conceive early in the breeding season is traditionally thought to be a function of pubertal status; however, recent evidence suggests that antral follicle count is positively associated with calving day in pubertal beef heifers. Additionally, there is evidence to suggest that the total number of ovarian follicles may alter response to hormonal synchronization protocols. The objectives of this study were to confirm the beneficial influence of antral follicle count on calving day in beef heifers and to determine whether response to PGF2α is associated with differences in antral follicle counts. A 2 × 2 factorial experiment was designed to investigate the interaction between calving period (early vs. late) and PGF2α (control vs. PGF) on calving day and antral follicle count in yearling beef heifers (n = 95). As intended, calving day was less in the early calving period compared to the late calving period (P < 0.01). There were no differences in calving day in response to treatment with PGF2α (P > 0.05). There was a significant interaction between calving period and PGF2α on age at first calving (P < 0.01), such that heifers treated with PGF2α that gave birth early were younger than heifers treated with saline that gave birth early. Calf weaning weights were greater in the early calving group than in the late calving group (P < 0.01). Heifers that gave birth in the early calving group possessed more antral follicles at prebreeding ultrasonographic examination than heifers that gave birth in the late calving group (P = 0.05). These findings confirm that antral follicle counts are associated with calving day in pubertal beef heifers. The use of antral follicle counts as a prebreeding phenotype provides additional utility to reproductive tract scoring for commercial production because of its association with calving day. As a prebreeding ovarian phenotype, antral follicle counts may hold additional applicability for organic beef producers looking to reduce the length of their calving season without increasing the total number of replacement heifers retained.

Glutamate Cysteine Ligase Modifier Subunit (Gclm) Null Mice Have Increased Ovarian Oxidative Stress and Accelerated Age-Related Ovarian Failure.

Glutathione (GSH) is the one of the most abundant intracellular antioxidants. Mice lacking the modifier subunit of glutamate cysteine ligase (Gclm), the rate-limiting enzyme in GSH synthesis, have decreased GSH. Our prior work showed that GSH plays antiapoptotic roles in ovarian follicles. We hypothesized that Gclm(-/-) mice have accelerated ovarian aging due to ovarian oxidative stress. We found significantly decreased ovarian GSH concentrations and oxidized GSH/oxidized glutathione redox potential in Gclm(-/-) vs Gclm(+/+) ovaries. Prepubertal Gclm(-/-) and Gclm(+/+) mice had similar numbers of ovarian follicles, and as expected, the total number of ovarian follicles declined with age in both genotypes. However, the rate of decline in follicles was significantly more rapid in Gclm(-/-) mice, and this was driven by accelerated declines in primordial follicles, which constitute the ovarian reserve. We found significantly increased 4-hydroxynonenal immunostaining (oxidative lipid damage marker) and significantly increased nitrotyrosine immunostaining (oxidative protein damage marker) in prepubertal and adult Gclm(-/-) ovaries compared with controls. The percentage of small ovarian follicles with increased granulosa cell proliferation was significantly higher in prepubertal and 2-month-old Gclm(-/-) vs Gclm(+/+) ovaries, indicating accelerated recruitment of primordial follicles into the growing pool. The percentages of growing follicles with apoptotic granulosa cells were increased in young adult ovaries. Our results demonstrate increased ovarian oxidative stress and oxidative damage in young Gclm(-/-) mice, associated with an accelerated decline in ovarian follicles that appears to be mediated by increased recruitment of follicles into the growing pool, followed by apoptosis at later stages of follicular development.

The effects of electromagnetic fields on the number of ovarian primordial follicles: An experimental study

The aim of this study was to evaluate the effect of an electromagnetic field (EMF), generated close to the ovaries, on primordial follicles. A total of 16 rats were used in this study. The study group consisted of rats exposed to an EMF in the abdominal region for 15 min/d for 15 days. Both the study and control group were composed of eight rats. After the treatment period of 15 days, the ovaries of the rats were extracted, and sections of ovarian tissue were taken for histological evaluation. The independent samples t test was used to compare the two groups. In the study group, the means of the right and left ovarian follicle numbers were 34.00 ± 10.20 and 36.00 ± 10.53, respectively. The average total ovarian follicle number was 70.00 ± 19.03. In the control group, the means of the right and left ovarian follicle numbers were 78.50 ± 25.98 and 71.75 ± 29.66, respectively, and the average total ovarian follicle number was 150.25 ± 49.53. The comparisons of the means of the right and left ovarian follicle numbers and the means of the total ovarian follicle numbers between the study and control groups indicated that the study group had significantly fewer follicles (p < 0.001, p = 0.011, and p = 0.002, respectively). This study found a significant decrease in the number of ovarian follicles in rats exposed to an EMF. Further clinical studies are needed to reveal the effects of EMFs on ovarian reserve and infertility.

Low numbers of ovarian follicles ≥3 mm in diameter are associated with low fertility in dairy cows

The total number of ovarian follicles ≥3mm in diameter (antral follicle count, AFC) during follicular waves varies among cattle of similar age, but AFC is highly repeatable within individuals. We hypothesized that lower AFC could be associated with reduced fertility in cattle. The AFC was assessed by ultrasonography for 2 d consecutively during the first wave of follicular growth of the estrous cycle, 4.6±1.43 d (mean ± SD) after estrus, in 306 Holstein-Friesian dairy cows approximately 70 d postpartum. Cows were classified into 3 groups based on AFC: low (AFC ≤15), intermediate (AFC=16 to 24), and high (AFC ≥25). During the cycle in which AFC was assessed and in subsequent cycles, cows were artificially inseminated (AI) following detection of estrus, and pregnancy status was assessed using ultrasonography. Cows with high AFC had 3.34 times greater odds of being pregnant at the end of the breeding season compared with cows with low AFC; the odds of a successful pregnancy at first service were 1.75 times greater in the intermediate compared with the low group. The predicted probability of a successful pregnancy by the end of the breeding period (length of breeding season was 86±16.3 d) was 94, 88, and 84% for the high, intermediate, and low AFC groups, respectively. No difference was evident among groups in 21-d submission rate (proportion of all cows detected in estrus and submitted for AI in the first 21 d of the breeding season), but the interval from calving to conception was shorter in the high (109.5±5.1 d) versus low (117.1±4 d) group, and animals with intermediate AFC received fewer services during the breeding season (2.3±0.1) compared with animals with low AFC (2.7±0.1). Lactating cows with ≤15 ovarian follicles have lower reproductive performance compared with cows with higher numbers of follicles, but the existence of a positive association between high numbers of ovarian follicles and fertility is yet to be established.

Assessment of fertility protection and ovarian reserve with GnRH antagonist in rats undergoing chemotherapy with cyclophosphamide

BackgroundReproductive function following chemotherapy is of increasing importance given that survival rates are improving. We assessed whether a gonadotropin-releasing hormone antagonist (GnRHant; cetrorelix) could promote ovarian protection against damage due to chemotherapy.MethodsForty-two female Wistar rats were used in this study. Animals were divided into four groups: group I (n = 9) received placebo twice; group II (n = 12) received placebo + cyclophosphamide (CPA); group III (n = 12) received GnRHant + CPA; and group IV (n = 9) received GnRHant + placebo. After medication, the estrous cycle was studied through vaginal smears. Rats were mated, pregnancy was documented and the number of live pups evaluated. Afterwards, rat ovaries were removed and prepared for histological studies. The ovarian cross-sectional area was measured and follicles were counted.ResultsCyclic changes in vaginal smears were observed in all but one animal after treatment, but group II had a significantly lower rate of animals with proestrus or estrus (p < 0.01). The offspring was markedly reduced by CPA treatment (group II, 3.00 +/- 1.33 pups vs. group I, 11.44 +/- 0.78 pups, p < 0.01) and this effect was partly reversed by pre-treatment with GnRHant (group III, 7.00 +/- 1.31 pups). The ovarian cross-sectional area was not significantly different between groups, neither was the number of individual follicle types. However, rats in Group IV had a higher total number of ovarian follicles than those in the control group (17.1 +/- 1.22 vs. 10.9 +/- 0.70, p < 0.05).ConclusionThe use of a GnRHant before CPA chemotherapy provided protection of fertility.

Increased ovarian follicle atresia in obese Zucker rats is associated with enhanced expression of the forkhead transcription factor FOXO1

It is well established that hyperinsulinemia, resulting from insulin resistance, plays a role in the pathophysiology of polycystic ovary syndrome (PCOS). The aim of this study was to investigate if ovarian follicular development and atresia are impaired in obese hyperinsulinemic (fa/fa) Zucker rats. To gain insight into the molecular mechanism of follicular atresia, we also examined the expression and localization of forkhead transcription factor FOXO1, a major regulator of cell fate decisions such as differentiation, cell-cycle arrest, and cell death. Serum insulin but not gonadotropin levels were significantly higher in obese (fa/fa) rats when compared to lean controls. Total ovarian follicle number and the percentage of atretic follicles were also significantly increased in obese (fa/fa) rats. Follicle atresia was associated with nuclear accumulation of FOXO1 transcription factor in TUNEL-positive granulosa cells. These results suggest a role for FOXO1 in granulosa cell apoptosis and increased ovarian follicle atresia associated with hyperinsulinemia.

Reproductive Fate Versus Environment

AMERICAN ASSOCIATION OF PHYSICAL ANTHROPOLOGISTS, 31 MARCH-4 APRIL 2009, CHICAGO, ILLINOIS