Evaluation of ovulation rate and ovarian phenotype in puberal heifers from a cattle population selected for increased ovulation rate

Long-term selection for increased ovulation rate (1984 to 2002) has resulted in a unique ovarian phenotype in the MARC Twinner cattle population. Ovulation rate and frequency of bilateral ovulations were examined by rectal palpation in 29,547 estrous cycles for 3,910 heifers (12 to 18 mo of age) in this population. Bilateral ovulations (one corpus lutuem [CL] on each ovary) were of interest because bilateral twin pregnancies result in decreased dystocia and increased calf survival. Ovulation rate increased linearly at a rate of 0.026 CL per year, and it currently averages 1.48 +/- 0.04 CL per estrous cycle. Concurrent with the increase in ovulation rate, the frequency of triplet ovulations increased from 0% to 2.3 +/- 0.8% (P < 0.001). Ovulation rate of both the right and left ovary increased equally at a rate of 0.013 CL per year, and mean ovulation rate of the right ovary remained greater than mean ovulation rate of the left ovary throughout the study (0.66 vs. 0.55 +/- 0.003 CL per estrous cycle; P < 0.001). Although correlations were low, ovulation rate of one ovary was negatively correlated (P < 0.001; r = -0.07) with ovulation rate of the same ovary in the previous estrous cycle, but positively correlated (P < 0.001; r = 0.13) with the contralateral ovary of the previous estrous cycle. The proportion of bilateral ovulations averaged 55.7 +/- 0.7%, a value greater than the predicted 49.5% (P < 0.001). In addition to dystocia and retained placenta, triplet pregnancies increase the incidence of pregnancies gestating fetuses of opposite sexes and subsequent incidence of freemartins; thus, selection pressure on ovulation rate may need to be adjusted in the MARC Twinner population. The proportion of bilateral ovulations in the population is greater than expected, and this may be an economically important trait, which will respond to selection and be beneficial for improving bovine reproductive efficiency. Understanding factors controlling the increased functional activity of the right ovary and bilateral ovulations may provide further insights into the mechanisms controlling follicle selection and methods to improve reproductive management of cattle.     

A dynamic model for ovulation rate reveals an effect of the estrogen receptor genotype on ovarian follicular development in the pig

A mathematical model that describes the recruitment and growth of ovarian follicles was fitted to data on ovulation rate and the measurements of plasma estradiol collected at times during the estrous cycle for individual gilts. The method of least squares was used to obtain estimates of the parameters of the mathematical model. The estimated model parameters were the maximum estradiol production for a follicle, development of each follicle after commitment, and a function describing the initial estradiol production of committed follicles. The estimated parameters for each pig were classified by estrogen receptor (ER) genotype (AA or BB) and analyzed using a multivariate analysis of variance. There were differences between genotypes (P < 0.05) for the parameter that described the initial distribution of individual follicles at recruitment. Gilts with ER genotype BB recruited follicles that varied more in size but had fewer very small follicles, indicating that the ER gene affects the relative estradiol secretion of the follicles at commitment. This analysis is an example of a general approach to genetic studies that uses a mathematical model of the physiology as a statistical basis for estimating gene action.     

Effect of selection for litter size and body weight on hormone-induced ovulation rate in mice.

Genetic differences in natural vs hormone-induced ovulation rates were compared in immature female mice from five lines that had undergone long-term single-trait and antagonistic index selection for litter size and(or) 6-wk BW. Lines used were control (K); high litter size (L+); high BW (W+); low litter size and high BW (L-W+); and high litter size and low BW (L+W-). Natural ovulation rate at a mean age of 34.3 d and hormone-induced (5 IU of pregnant mare's serum gonadotropin followed 2 d later by 5 IU of human chorionic gonadotropin) superovulation rate at a fixed age of 31 d were obtained. Total number of eggs ovulated was affected by line (P less than .001), treatment (P less than .001), and line x treatment interaction (P less than .001). Line differences were subsequently tested within treatment because of the significant line x treatment interaction. Line differences were important (P less than .001) for natural ovulation, hormone-induced ovulation, and response to hormones. Mean natural ovulation rates for K, L+, W+, L-W+, and L+W- were 14.1, 19.8, 15.1, 13.6, and 16.4, respectively. Selection changed ovulation rate by 40, 16, 7, and -4% in the L+, L+W-, W+ and L-W+ lines, respectively (P less than .01). Hormone-induced ovulation rates in K, L+, W+, L-W+, and L+W- were 32.3, 24.6, 19.6, 20.9, and 22.1, respectively. Exogenous hormones increased ovulation by 18.2, 4.8, 4.6, 7.3, and 5.7 ova for K, L+, W+, L-W+, and L+W-, respectively (P less than .001). Lines with lower natural ovulation rates had higher responses to superovulation. Increased ovulation rate due to treatment ranged from 24.3% in L+ to 129% in K. These results indicate significant differences among lines in ovarian response to exogenous hormones.     

Selection for ovulation rate in rabbits

An experiment of selection for ovulation rate was carried out. Animals were derived from a synthetic line first selected 12 generations for litter size, then 10 generations for uterine capacity. Selection was relaxed for 6 generations. Selection was based on the phenotypic value of ovulation rate with a selection pressure on does of 30%. Males were selected from litters of does with the highest ovulation rate. Males were selected within sire families in order to reduce inbreeding. Ovulation rate was measured in the second gestation by a laparoscopy, 12 days after mating. Each generation had about 80 females and 20 males. Results of three generations of selection were analyzed using Bayesian methods. Marginal posterior distributions of all unknowns were estimated by Gibbs sampling. Heritabilities of ovulation rate (OR), number of implanted embryos (IE), litter size (LS), embryo survival (ES), fetal survival (FS), and prenatal survival (PS) were 0.44, 0.32, 0.11, 0.26, 0.35, and 0.14, respectively. Genetic correlation between OR and LS was 0.56, indicating that selection for ovulation rate can augment litter size. Response to selection for OR was 1.80 ova. Correlated responses in IE and LS were 1.44 and 0.49, respectively. Selection for ovulation rate may be an alternative to improve litter size.     

TRAIL-decoy receptor 1 plays inhibitory role in apoptosis of granulosa cells from pig ovarian follicles

Previously, we histochemically examined the localization of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and its receptors in porcine ovarian follicles, and demonstrated a marked reduction in the expression of TRAIL-decoy receptor-1 (DcRI) in granulosa cells of atretic follicles. In the present study, to confirm the inhibitory activity of DcR1 in granulosa cells, granulosa cells prepared from healthy follicles were treated with phosphatidylinositol-specific phospholipase C (PI-PLC) to cleave glycophospholipid anchor of DcR1 and to remove DcR1 from the cell surface, and then incubated with TRAIL. PI-PLC treatment increased the number of apoptotic cells induced by TRAIL. The present finding indicated the possibility that TRAIL and its receptors were involved in induction of apoptosis in granulosa cells during atresia, and that DcR1 plays an inhibitory role in granulosa cell apoptosis.     

Plasma LH and progesterone levels before and after ovulation and observation of ovarian follicles by ultrasonographic diagnosis system in dogs

Recently, canine frozen semen has been attracting attention for breeding purposes, and methods of judging ovulation and optimum timing for insemination have become important. As methods of predicting the canine ovulation, vaginal smear, plasma sex hormone levels and ultrasonographic diagnosis system (US) have been investigated in combination, but a standard technique has not yet been established. Therefore, we investigated a method of predicting canine ovulation in dogs by US, and by measuring plasma LH and progesterone (P) levels three times a day. Ovulation could be observed by detecting irregularly shaped ovarian follicles by US in six of 11 dogs (54.5%). In these dogs, the time between the LH peak and ovulation was 24-48 hr, 38.0 hr on average. The P level on the ovulation day was 1.88-2.81 ng/ml, 2.34 ng/ml on average. A value of 1.88 ng/ml was detected in one dog, but the other five dogs showed P levels of 2 ng/ml or higher. The P level on the day before ovulation was 0.8-1.56 ng/ml, 1.12 ng/ml on average. Assuming that ovulation occurred two days after the LH peak in the 11 experimental dogs, the P level was 2.12-4.06 ng/ml, 2.78 ng/ml on average. The period of a high LH level, not less than 10 ng/ml, continued for 12 hr around the LH peak. Based on these findings, to predict ovulation using US and LH level, it would be necessary for the tests to be performed several times a day. In contrast, it was shown that the day on which a plasma P level of 2 ng/ml or higher was detected by the test performed once a day corresponded to the ovulation day.     

Changes in fetal organ weights during gestation after selection for ovulation rate and uterine capacity in swine

We hypothesized that the ability of the fetus to alter nutrient shunting and organ growth might be associated with uterine capacity. White crossbred gilts from a randomly selected control line, a line selected for ovulation rate, and a line selected for uterine capacity (UC) were unilaterally hysterectomized-ovariectomized at 160 d of age, mated at estrus, and slaughtered at 45, 65, 85, and 105 d of gestation (9 to 18 gilts for each line x day combination). Analysis of the data revealed that heart weights and fetal weights were decreased in the ovulation rate line. No significant differences were obtained in fetal, placental, or fetal organ weights between the control and UC lines. Allometric growth of organs was assessed by examination of the slopes of the relationships between fetal weights and fetal organ weights after natural log transformation. Only the relative growth of the liver differed between selection lines and was greater (P = 0.01) in the UC compared with the control line during early pregnancy (d 45 and 65). Allometric growth of the fetal brain, liver, and heart differed with day of gestation. A brain-sparing effect was greater (P < 0.01) on d 85 and 105 compared with d 45 and 65. By contrast, a heart-sparing effect was present during early gestation and disappeared in later gestation. Fetal liver weights were hypersensitive to differences in fetal weights on d 45, possibly associated with placental effects on fetal liver weight. Fetal spleen weights were proportional to fetal weights throughout gestation. These results indicate that selection for ovulation rate decreased total fetal and fetal heart weights, and that selection for UC altered the relationship between total fetal and fetal liver weights during early gestation. Results further indicate significant changes in allometric growth of organs during gestation.     

Direct responses to selection for increased litter size, decreased age at puberty, or random selection following selection for ovulation rate in swine

Nine generations of selection for high ovulation rate were followed by two generations of random selection and then eight generations of selection for increased litter size at birth, decreased age at puberty, or continued random selection in the high ovulation rate line. A control line was maintained with random selection. Line means were regressed on generation number and on cumulative selection differentials to estimate responses to selection and realized heritabilities. Genetic parameters also were estimated by mixed-model procedures, and genetic trends were estimated with an animal model. Response to selection for ovulation rate was about 3.7 eggs. Response in litter size to selection for ovulation rate was .089 +/- .058 pigs per generation. Average differences between the high ovulation rate and control lines over generations 10 to 20 were 2.86 corpora lutea and .74 pigs (P less than .05). The regression estimate of total response to selection for litter size was 1.06 pigs per litter (P less than .01), and the realized heritability was .15 +/- .05. When the animal model was used, the estimate of response was .48 pigs per litter. Total response in litter size to selection for ovulation rate and then litter size was estimated to be 1.8 and 1.4 pigs by the two methods. Total response to selection for decreased age at puberty was estimated to be -15.7 d (P less than .01) when data were analyzed by regression (realized heritability of .25 +/- .05) and -17.1 d using the animal model. No changes in litter size occurred in the line selected for decreased age at puberty. Analyses by regression methods and mixed-model procedures gave similar estimates of responses and very similar estimates of heritabilities.     

Oxygen consumption rate of early pre-antral follicles from vitrified human ovarian cortical tissue

The study of human ovarian tissue transplantation and cryopreservation has advanced significantly. Autotransplantation of human pre-antral follicles isolated from cryopreserved cortical tissue is a promising option for the preservation of fertility in young cancer patients. The purpose of the present study was to reveal the effect of vitrification after low-temperature transportation of human pre-antral follicles by using the oxygen consumption rate (OCR). Cortical tissues from 9 ovaries of female-to-male transsexuals were vitrified after transportation (6 or 18 h). The follicles were enzymatically isolated from nonvitrified tissue (group I, 18 h of transportation), vitrified-warmed tissue (group II, 6 and 18 h of transportation) and vitrified-warmed tissue that had been incubated for 24 h (group III, 6 and 18 h of transportation). OCR measurement and the LIVE/DEAD viability assay were performed. Despite the ischemic condition, the isolated pre-antral follicles in group I consumed oxygen, and the mean OCRs increased with developmental stage. Neither the transportation time nor patient age seemed to affect the OCR in this group. Meanwhile, the mean OCR was significantly lower (P < 0.05) in group II but was comparable to that of group I after 24 h of incubation. The integrity of vitrified-warmed primordial and primary follicles was clearly corroborated by the LIVE/DEAD viability assay. These results demonstrate that the OCR can be used to directly estimate the effect of vitrification on the viability of primordial and primary follicles and to select the viable primordial and primary follicles from vitrified-warmed follicles.     

Plasma gonadotropins and ovarian hormones during the estrous cycle in high compared to low ovulation rate gilts

Mature gilts classified by low (12 to 16 corpora lutea [CL], n = 6) or high (17 to 26 CL, n = 5) ovulation rate (OR) were compared for plasma follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone, estradiol-17beta, and inhibin during an estrous cycle. Gilts were checked for estrus at 8-h intervals beginning on d 18. Blood samples were collected at 8-h intervals beginning on d 18 of the third estrous cycle and continued for one complete estrous cycle. Analysis for FSH and LH was performed on samples collected at 8-h intervals and for ovarian hormones on samples collected at 24-h intervals. The data were standardized to the peak of LH at fourth (d 0) and fifth estrus for the follicular phase and analyzed in discrete periods during the periovulatory (-1, 0, +1 d relative to LH peak), early-luteal (d 1 to 5), mid-luteal (d 6 to 10), late-luteal (11 to 15), periluteolytic (-1, 0, +1 d relative to progesterone decline), and follicular (5 d prior to fifth estrus) phases of the estrous cycle. The number of CL during the sampling estrous cycle was greater (P < 0.005) for the high vs low OR gilts (18.8 vs 14.3) and again (P < 0.001) in the cycle subsequent to hormone measurement (20.9 vs 14.7). For high-OR gilts, FSH was greater during the ovulatory period (P = 0.002), the mid- (P < 0.05) and late-luteal phases (P = 0.01), and tended to be elevated during the early-luteal (P = 0.06), but not the luteolytic or follicular periods. LH was greater in high-OR gilts during the ovulatory period (P < 0.005), but not at other periods during the cycle. In high-OR gilts, progesterone was greater in the mid, late, and ovulatory phases (P < 0.005), but not in the follicular, ovulatory, and early-luteal phases. Concentrations of estradiol-17beta were not different between OR groups during the cycle. Inhibin was greater for the high OR group (P < 0.005) during the early, mid, late, luteolytic, and follicular phases (P < 0.001). The duration of the follicular phase (from last baseline estrogen value to the LH peak) was 6.5 +/- 0.5 d and was not affected by OR group. These results indicate that elevated concentrations of both FSH and LH are associated with increased ovulation rate during the ovulatory phase, but that only elevated FSH during much of the luteal phase is associated with increased ovulation rate. Of the ovarian hormones, both inhibin and progesterone are highly related to greater ovulation rates. These findings could aid in understanding how ovulation rate is controlled in pigs.     

Direct responses to six generations of selection for ovulation rate or prenatal survival in Large White pigs

Effects of selection for ovulation rate or prenatal survival were examined using data from 3 pigs lines derived from the same base Large White population. Two lines were selected for 6 generations on high ovulation rate at puberty (OR line) or high prenatal survival corrected for ovulation rate in the first 2 parities (PS line). The third line was an unselected control line. Genetic parameters for ovulation rate on the left, right, and both ovaries at puberty (ORPL, ORPR, and ORP, respectively) and at fertilization (ORFL, ORFR, and ORF, respectively), total number of piglets born (TNB) per litter, prenatal survival (PS = TNB/ORF), and PS corrected for ovulation rate (CPS = PS + 0.018ORF) were estimated using REML methodology. Responses to selection were estimated by computing differences between OR or PS and control lines at each generation using least squares and mixed models methodology. Average genetic trends were computed by regressing line differences on generation number. Realized heritabilities were estimated using standard procedures. Heritability estimates were 0.17, 0.11, 0.34, 0.13, 0.09, 0.33, 0.14, 0.11, and 0.17 (SE = 0.01 to 0.03) for ORPL, ORPR, ORP, ORFL, ORFR, ORF, PS, CPS, and TNB, respectively. Realized heritabilities were 0.37 +/- 0.08 and 0.10 +/- 0.09 for ORP and CPS, respectively. The different measures of ovulation rate had strong genetic correlations (r(g) > 0.7). The ORF had midrange negative genetic correlations with PS and CPS (-0.45 +/- 0.07 and -0.42 +/- 0.08, respectively). The ORP also had an antagonistic genetic relationship with PS (-0.26 +/- 0.07) but was almost independent from CPS (-0.02 +/- 0.11). The TNB was moderately correlated with ORP and ORF (r(g) = 0.41 +/- 0.09 for both traits). Average genetic trends in OR and PS lines were, respectively, 0.49 +/- 0.10 and 0.11 +/- 0.10 for ORP, and 0.43 +/- 0.11 and 0.11 +/- 0.11 for ORF. Responses to selection were slightly superior in the left than in the right ovary. No significant difference was found for PS or CPS in any of the lines. The TNB did not change in the OR line but significantly improved in the PS line (0.24 +/- 0.11 piglets/generation).     

Role of VEGF in pig preantral follicles

Veterinary Research Communications -     

Control of ovarian follicles activity in the ewe.

During the ovine estrous cycles, three waves of follicular growth, closely associated with the FSH secretion pattern, were observed. The parameters of these follicular waves and the ability of follicles to produce steroids in vitro were studied in various conditions. In vivo, the follicular events were similar between the breeding season and the anestrus, except for the lack of ovulation; but at the end of the breeding season and in anestrus, the follicles lose a big part of their aromatization ability. In ewes carrying the Booroola fecundity gene or Cambridge fecundity gene, the reduction in follicular atresia seems to be one of the main follicular features implicated in the control of high ovulation rate. In vitro, the most relevant difference is an early acquisition of estrogen production ability of small follicles in Booroola fecundity gene barring ewes. Fluoro-gestone-acetate (FGA) pessaries reduced the number of growing follicles; despite this effect disappearing after the sponge withdrawal, the ovulation rate is significantly reduced. But an equine chorionic gonadotrophin (eCG) treatment restores the ovulation rate (OR) by reducing the atresia rate of pre-ovulatory follicles. In similar conditions, a pretreatment of the ewes with melatonin again reduced the atresia rate of large follicles and resulted in an increased ovulation rate. In vitro, FGA blocked aromatization ability, and melatonin inhibited both androstenedione and estradiol production, but a further treatment with eCG partly restores the steroid secretion. Immunization against androstenedione leads to a higher OR, owning to a reduced atresia of large follicles. Daily growth hormone injections for a hole cycle resulted in an increased follicular population and ovulation rate, while FSH plasma levels decreased and the follicle sensitivity to gonadotrophins was reduced.     

Direct and correlated responses to two-stage selection for ovulation rate and number of fully formed pigs at birth in swine

Our objectives were to estimate responses and genetic parameters for ovulation rate, number of fully formed pigs at birth, and other production traits following two-stage selection for increased ovulation rate and number of fully formed pigs. Eight generations of selection were practiced in each of two lines. One selection line was derived from a line that previously selected eight generations for an index to increase ovulation rate and embryonic survival (the IOL pigs). The other selection line was derived from the unselected control line of the index selection experiment (the COL pigs). The control line (C) was continued with random selection. Due to previous selection, Line IOL had greater ovulation rate (4.24 +/- 0.38 and 4.14 +/- 0.29 ova) and litter size (1.97 +/- 0.39 and 1.06 +/- 0.38 pigs) at Generation 0 of two-stage selection than did Lines COL and C. In Stage 1, all gilts from 50% of the largest litters were retained. Approximately 50% of them were selected for ovulation rate in Stage 2. Gilts selected for ovulation rate were mated to boars selected from the upper one-third of the litters for litter size. At Generations 7 and 8, differences in mean EBV for ovulation rate and litter size between Lines IOL and C were 6.20 +/- 0.29 ova and 4.66 +/- 0.38 pigs; differences between Lines COL and C were 2.26 +/- 0.29 ova and 2.79 +/- 0.39 pigs; and differences between Lines IOL and COL were 3.94 +/- 0.26 ova and 1.86 +/- 0.39 pigs. Regressions of line mean EBV on generation number were 0.27 +/- 0.07 ova and 0.35 +/- 0.06 pigs in Line IOL; 0.30 +/- 0.06 ova and 0.29 +/- 0.05 pigs in Line COL; and 0.01 +/- 0.07 ova and 0.02 +/- 0.05 pigs in Line C. Correlated responses were decreased age at puberty and increased number of pigs born alive, number of mummified pigs, prenatal loss, and individual and litter birth weight. Two-stage selection for ovulation rate and number of pigs per litter is a promising procedure to improve litter size in swine.     

Induction of ovarian cystic follicles in sheep

Cystic follicles are a significant cause of infertility in women, dairy cattle and sheep. Sheep were used as a model to identify factors that may elicit formation of cystic follicles. Insulin resistance and elevated LH activity were tested in overweight ewes because of associations among these factors and the formation of cystic follicles. Sheep were synchronized using a progesterone-releasing pessary and insulin resistance was induced during the synchronization period through administration of bovine somatotropin. Following removal of pessaries follicular growth was stimulated by treatment with eCG or eCG and hCG (PG-600). Follicular growth was monitored via daily transrectal ultrasonography and blood samples were collected for hormonal analyses. Six of 18 ewes had a subnormal or absent preovulatory gonadotropin surge and developed cystic follicles. Neither insulin resistance nor elevated LH activity were associated with formation of cystic follicles. Ewes that developed cystic follicles were heavier (93 +/- 4 kg) than ewes that ovulated (81 +/- 3 kg; P = 0.02). Furthermore, following pessary removal and initiation of daily ultrasonography, ewes that developed cystic follicles lost body weight (-3 +/- 1%), while ovulatory ewes continued to gain body weight (1 +/- 1%; P = 0.005). It is speculated that in heavy ewes metabolic factors associated with acute body weight loss inhibit the positive feedback of estradiol and thereby suppress the preovulatory gonadotropin surge leading to formation of cystic follicles.     

Role for LDL in estradiol-synthesis capacity of bovine ovarian follicles

The steroidogenic capacity of several tissues has been shown to be regulated by lipoproteins. However, the ability of the various lipoprotein classes to affect steroid production in general and estradiol-synthesis in particular has not been established in bovine ovarian cells. In previous studies, we described alterations in the lipoprotein profiles in follicles of different sizes and corresponding changes in lipoprotein-receptor gene expression. In the present study, the effect of low-density lipoprotein (LDL)-enriched medium on aromatization competence of whole ovarian follicles was determined in vitro. Gene expression of aromatase increased and that of selective-uptake receptors decreased in the presence of LDL. These results suggest a role for LDL availability in folliculogenesis.