Changes in the cumulus-oocyte complex of subordinate follicles relative to follicular wave status in cattle

We investigated factors that affect cumulus-oocyte complex (COC) morphology and oocyte developmental competence in subordinate follicles on different days after follicular wave emergence in beef heifers. In Experiment 1, heifers (n = 13) were assigned at random to COC aspiration during the growing/static (Days 1 to 3) or regressing (Day 5) phase of subordinate follicle development (follicular wave emergence = Day 0). Follicular wave emergence was induced by transvaginal ultrasound-guided follicular ablation, ovaries were collected at slaughter, all follicles > or = 2 mm except the dominant follicle were aspirated, and COC were microscopically evaluated for morphology. There was a greater percentage of COC with expanded cumulus layers on Day 5 (42.4%) than on Days 1 to 3 (2.2%). In Experiment 2, heifers (n = 64) at random stages of the estrous cycle had all follicles > or = 5 mm ablated and 4 d later, 2 doses of PGF were injected 12 h apart; heifers were monitored daily by ultrasonography for ovulation (Day 0 = follicular wave emergence). Heifers were assigned to the following time periods for oocyte collection from subordinate follicles: Days 0 and 1 (growing phase), Days 2, 3 and 4 (static phase), and Days 5 and 6 (regressing phase). Ovaries were individually collected at slaughter, and all follicles > or 2 mm except for the dominant follicle were aspirated. The COC were morphologically evaluated and then matured, fertilized and cultured in vitro. Expanded COC were more frequent during the regressing phase (53.4%) than the growing or static phase (14.4 and 17.8%, respectively; P < 0.05). While the proportions of COC with > or = 4 layers of cumulus cells and denuded oocytes were higher (P < 0.05) in the growing and static phases, the production of morulae was highest (P < 0.05) with COC collected from subordinate follicles during the regressing phase. In Experiment 3, heifers (n = 18) were assigned at random to oocyte collection from subordinate follicles 3 and 4 d (static phase) or 5 and 6 d (regressing phase) after follicular wave emergence. The heifers were monitored ultrasonically for ovulation (Day 0 = follicular wave emergence); COC were collected from all follicles (> or = 5 mm) except for the dominant follicle by transvaginal ultrasound-guided follicle aspiration 3 to 6 d later. Recovered oocytes were stained and examined microscopically to evaluate nuclear maturation. A higher proportion of oocytes collected on Days 5 and 6 showed evidence of nuclear maturation (50%) than on Days 3 and 4 (8.3%; P < 0.05). Results support the hypothesis that COC morphology and oocyte developmental competence change during the growing, static and regressing phases of subordinate follicle development.     

Morphology and developmental competence of bovine oocytes relative to follicular status

In cattle, follicle dimension has been used as the main criterion for selection of oocytes for in vitro embryo production. However, follicles with similar diameters may be in very different physiologic phases. The aim of this study was to investigate whether morphology and developmental competence of cumulus-oocyte complexes (COCs) are related to the phase of development of the follicle, and presence of the corpus luteum (CL) or the dominant follicle in the ovary from which the COCs were collected. Cows (n = 143) were given a luteolytic dose of PGF(2alpha) and 8 days later underwent transvaginal ultrasound guided ablation of follicles > or =4mm to induce emergence of a new follicular wave. Cows (n = 10-20 per replicate) were slaughtered on Day 2, 3, 5 or 7 (Day 0 = follicular wave emergence), equivalent to the growing, early static, late static, and regressing phases of subordinate follicle development. COCs were collected from subordinate follicles > or =3mm, were classified as denuded, degenerated or healthy, and underwent IVM-IVF-IVC. The proportion of oocytes that developed to the blastocyst stage was higher (P<0.05) in those collected on Day 5 after wave emergence (23%) than on Day 2 (12%), 3 (13%) or 7 (16%). Data did not support the hypothesis of a local effect of the CL or dominant follicle. We conclude that a positive relationship exists between early follicular regression and oocyte competence. Moreover, morphologic characteristics of oocyte quality used in this study were not predictive in identifying competent oocytes.     

Morphological classification of the ovaries in relation to the subsequent oocyte quality for IVF-produced bovine embryos

Although some inferences have been made regarding the morphological aspects of the ovaries in relation to the subsequent oocyte developmental competence in an in vitro system, the influence of ovarian morphology, taken as a pair, has yet to be demonstrated. The present study addresses this limitation. Forty pairs of ovaries from 5 morphological classes were examined to determine whether their characteristics could influence oocyte yield and developmental competence in vitro. An ovary was designated as bearing a corpus luteum (CL) with a dominant follicle (DF) a cyst (CY) or none of these structures (NO). Thus, the paired classes considered in this study consisted of 1) CL-NO 2) CL-DF 3) CL + DF-NO 4) NO-DF and 5) NO-NO. Comparisons were made among the members of 3 subgroups CL, NO and DF. Within the CL-subgroup, the pairs of CL-NO ovaries resulted in higher (P < 0.01) number of oocytes, cleavage rates and blastocyst formation per ovary than in the other categories (CL + DF-NO and CL-DF), with the latter being superior (P < 0.01) to that of CL + DF-NO in terms of cleavage rates. In the NO-subgroup, NO-CL pairs yielded higher (P < 0.01) rates of oocyte recovery and cleavage than the NO-DF pairs, and the latter was inferior (P < 0.05) to that of NO-NO ovaries for the 2 indices. Further, blastocyst rates from the NO-CL pairs was higher (P < 0.01) compared with those of NO-CL + DF, NO-DF, and NO-NO groups. And, in the DF-subgroup, the DF-CL pairs gave a higher (P < 0.05) oocyte yield and cleavage rate (P < 0.01) than the pairs of DF-NO ovaries but not significantly different in blastocyst formation. The overall oocyte recovery, cleavage and blastocyst rates for the 5 classes were, in a decreasing order CL-NO; NO-NO; CL-DF; CL + DF-NO; and DF-NO. Our results suggest that the morphological classification of ovarian pairs could be a useful means for predicting the developmental competence of oocytes in vitro, and that the presence of a dominant follicle in either one or both ovaries of a pair has a negative effect on the IVF-produced bovine embryos.     

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.     

Influence of the dominant follicle on oocytes from subordinate follicles

As the oocyte grows within the follicle, a number of factors influence its health and developmental competence. These factors include follicle size, day of estrous cycle, level of atresia and influence of other follicles such as the dominant follicle. Follicles were dissected from ovaries of synchronized dairy cows on four days during the estrous cycle, and the oocyte from each follicle collected, matured, fertilized and cultured singly until Day 8. Development to blastocyst was greater in oocytes collected during phases of follicular growth than those collected during phases of follicular dominance (P<0.001) over all follicle size categories. Oocyte competence tended to increase with increasing follicle size (P<0.1). Follicular cells analyzed by flow cytometry showed an increase in proportion of apoptotic cells in subordinate follicles during the dominant phase compared to growth phase (P<0.05). Thus, the dominant follicle on both oocyte competence and levels of atresia. Further studies on the effect of dominance has shown that lactate production in cumulus-oocyte-complexes (COCs) from medium-sized follicles collected during a dominance phase and small follicles collected during a growth phase are no different from other follicles, despite having significantly lower uptake of glucose (P<0.1). Thus, COCs from different follicle subclasses differ in their nutrient requirements, and current IVM technology needs further improvement to better assist those oocytes that are developmentally challenged.     

Intraovarian relationships among dominant and subordinate follicles and the corpus luteum in heifers

Previous studies demonstrated that waves of follicular activity develop approximately every 9 d in cattle during the estrous cycle and early pregnancy. A dominant follicle develops from each wave and the remaining follicles (subordinates) begin to regress after a few days. In this study, intraovarian luteal and follicular interrelationships were examined during the follicular waves of the estrous cycle and pregnancy using data obtained by ultrasonography. During the estrous cycle, no intraovarian relationships were found between the ovary containing the corpus luteum and the ovary containing the dominant follicle (n = 165), or between the location of the corpus luteum and the characteristics of the dominant follicle. During pregnancy, however, the frequency distribution for the number of follicular waves with the dominant follicle and corpus luteum on the same or opposite ovaries differed (P<0.05) among Waves 1 to 10. The two structures (dominant follicle and corpus luteum) were more often in opposite ovaries during Waves 3 to 10 (combined frequency, 75%) than during Waves 1 and 2. During pregnancy, dominant follicles of consecutive waves differed (P<0.05) among Waves 1 to 8 in the frequency with which they appeared in the same versus the opposite ovary. The difference seemed primarily due to an increased frequency of consecutive follicles on the same ovary for Waves 4 to 8 (combined frequency, 80%). During both the estrous cycle and pregnancy, there was no significant intraovarian effect of the dominant follicle on the day of detection of the next dominant follicle, on the growth rate of the largest subordinate follicle, or on the length of the interval from wave origin to cessation of growth of the largest subordinate; these results indicate that previously postulated suppressive effects between follicles are exerted through systemic channels.     

Factors affecting number of surface ovarian follicles and oocytes yield and quality in Egyptian buffaloes

Three experiments were conducted to evaluate factors affecting number of surface ovarian follicles and oocytes yield and quality in buffalo. In Experiment 1, ovaries (n = 126) were collected in pairs from slaughtered anoestrus, early pregnant and cyclic buffaloes. Ovarian follicles (1-3, 4-9 and > or = 10 mm diameter) were counted, aspirated and oocytes were recovered and evaluated. In Experiment 2, ovaries were divided into 2 groups. Group 1, ovaries bearing a CL (n = 74) and Group 2 non-bearing CL (n = 74), ovarian follicles (2-8 mm) were counted, aspirated and oocytes evaluated. In Experiment 3, oocytes were recovered using aspiration or slicing methods. In all experiments, oocytes were classified into good, fair, poor and denuded. Results showed that the development of small and total ovarian follicles are continuous and independent in early pregnant or cyclic buffalo cows, however, it significantly decreased (P < 0.01) in the ovaries of anoestrus buffaloes. Number of medium and large size follicles was significantly increased (P < 0.01) in cyclic buffaloes on Days 10-16 and 17-22 of oestrous cycle, while large follicles was significantly decreased (P < 0.01) in the ovaries of pregnant buffaloes. A significantly higher (P < 0.01) percentage of poor and denuded oocytes were recovered from ovaries of anoestrus and pregnant buffalo. While, the highest (P < 0.01) percentage of good quality oocytes were recovered from ovaries of cyclic buffaloes on Days 1-3 and 10-16 of oestrous cycle, eliciting that the stage of oestrous cycle is affecting the quality of buffalo oocytes. In addition, the presence of a CL stimulates the development of a significantly higher (P < 0.01) number ovarian follicles which produced a significantly higher (P < 0.05) number of good quality oocytes. Slicing of buffalo ovaries produced a significantly higher number of fair, poor and denuded oocytes. In conclusion, number of ovarian follicles and yield and quality of oocytes were affected by the reproductive status, stage of the oestrous cycle, presence of a CL and the method of oocytes retrieval.     

Kinase pathways in dominant and subordinate ovarian follicles during the first wave of follicular development in sheep

The mechanism by which one or more dominant ovarian follicles continue development while other subordinate follicles regress is not known. The mitogen activated protein kinases (MAPKs) are a group of kinases that are activated by hormonal factors and form a cascade of processes that regulate cell growth, division and differentiation. The aim of the present experiment was to characterise the presence of the MAPKs, Erk 1/Erk 2 and Akt in healthy dominant follicles and regressing subordinate follicles. Following in vivo monitoring of ovarian follicle development, three ewes were ovariectomised and the follicular fluid and follicle wall (theca and granulosa cells) saved from the dominant and largest subordinate follicle. The dissected diameter and follicular fluid oestradiol concentration of the dominant follicle was larger (P<0.01) than the largest subordinate follicle (6.5+/-0.0mm and 41.3+/-4.9ng/ml versus 4.7+/-0.3mm and 0.6+/-0.4ng/ml). Western blot analyses showed that there was more Akt (202.7+/-6.4 versus 59.6+/-32.7 units; P<0.05) and Erk 1/Erk 2 (104.5+/-10.6 versus 0.3+/-0.2 units; P<0.01) present in follicle wall samples from the dominant compared to the largest subordinate follicles. Phosphorylated forms of Akt and Erk 1/Erk 2 were detected in samples from dominant but not subordinate follicles. We suggest that signal transduction pathways involving Akt and Erk 1/Erk 2 may play an important role in determining the outcome of ovarian follicle growth and development in sheep.     

Utilization of the growth phase of the first follicular wave for bovine oocyte collection improves blastocyst production

Characteristics of the follicle population and oocyte developmental competence at selected stages of follicular development were studied in cows with the aim to increase embryo production derived from oocytes collected by transvaginal aspiration. In Experiment 1, the growth phase before dominant follicle selection and the low dominant phase during dominant follicle regression were compared. Twenty-four cyclic Holstein cows, 4 to 6 yr of age, were divided into 2 groups. Animals were synchronized using two injections of prostaglandin F2alpha at 11 d intervals, and onset of estrus was determined (Day 0). Using ultrasonography, all follicles were counted and classified. Oocytes were aspirated once on Days I through 3 (Group 1, n=5) or Days 15 and 16 (Group 2, n=3) of the estrus cycle. The experiment was carried out in 3 replicates. In Experiment 2, the growth phase of the first follicular wave before dominant follicle selection was characterized in detail. Twelve cows of the same breed and age were divided into 3 groups. Their first estrus was synchronized as in Experiment 1, and each following estrus was induced using one injection of prostaglandin F2alpha administered 4 to 6 d after each aspiration performed. The ovaries were examined, and oocytes were collected repeatedly (total of 5 times per cow) on Days 1 (Group 3, n=4), 2 (Group 4, n=4) or 3 (Group 5, n=4) after estrus at 10 d intervals during a 40 d period. Viable oocytes were matured, fertilized and cultured using the standard methods. In Experiment 1, the mean numbers (+/-SD) of all follicles and of recovered and viable oocytes per donor were higher in Group 1 than in Group 2, but only the mean numbers (+/-SD) of larger follicles and recovered oocytes were statistically significant (8.0 +/- 0.6 and 6.2 +/- 0.6.vs. 3.3 +/- 0.5 and 2.8 +/- 0.2; P< 0.05). In Experiment 2, the percentage of larger follicles out of all visible follicles and the mean numbers (+/-SD) of larger follicles per donor were significantly higher (P<0.05) in Groups 4 (75.7 and 9.1 +/- 2.7) and 5 (66.3 and 8.5 +/- 2.9) when compared to Group 3 (27.9 and 3.8 +/- 0.8). The development rate of fertilized oocytes was significantly higher (P<0.05) in Groups 4 (27.8) and 5 (27.5) than in Group 3 (12.8). It can be concluded that it is possible to improve the efficiency of transvaginal aspiration and in vitro embryo production by utilization of the growth phase of the first follicular wave before dominant follicle selection.     

Oocyte retrieval and histological studies of follicular population in buffalo ovaries

Ovaries (N = 250) from slaughtered buffaloes were collected to study follicular population and compare methods of oocyte retrieval. The number and size of surface follicles were recorded and grouped into different categories. Different sized follicles in relation to oocyte diameter were studied histologically. Yield of oocytes per ovary were less (P < 0.05) from ovaries bearing a corpus luteum (CL). Techniques used for oocyte recovery included slicing, follicle puncture and aspiration. The oocyte recovery rate was greatest (P < 0.05) using slicing. The average number of visible surface follicles was 5.20 ± 0.97 with mean numbers of 2.5, 1.2, 0.82 and 0.62 per ovary for follicles sized 4, 8, 12 and 12 mm respectively. Histological studies revealed large numbers of primordial follicles in prepubertal and atretic follicles in senile buffaloes. They also established a biphasic relationship of growth between oocyte diameter and follicular size.     

Evaluation of antral follicle growth in the macaque ovary during the menstrual cycle and controlled ovarian stimulation by high‐resolution ultrasonography

To date, ultrasonography of monkey ovaries is rare and typically of low resolution. The objectives of this study were to use state‐of‐the‐art, high‐resolution, transabdominal ultrasonography with real‐time Doppler capabilities to: (1) determine whether one can reliably detect in real time the large dominant follicle, the corpus luteum (CL), and small (<2 mm) antral follicles on the ovaries of rhesus monkeys during the natural menstrual cycle; and (2) predict the follicular response of rhesus ovaries to controlled ovarian stimulation (COS) protocols. Rhesus monkeys were selected for transabdominal ultrasonography using a GE Voluson 730 Expert Doppler System at discrete stages of the menstrual cycle. Subsequently, serial ultrasound scanning was employed to observe growth of antral follicles and the CL. Finally, females were scanned to assess follicular growth during COS. The dominant structure and small antral follicles (<2 mm) were reliably visualized in real time. The follicle destined to ovulate could be identified by size differential by day 3 of the follicular phase. The number of small antral follicles present before onset of COS protocol correlated positively with the number of metaphase II‐stage oocytes collected after treatment. The results of this study demonstrate that the population dynamics of antral follicle pools can be noninvasively evaluated in monkeys during natural and pharmacologic ovarian cycles. Am. J. Primatol. 71:384–392, 2009. © 2009 Wiley‐Liss, Inc.     

The relationship of follicle atresia to follicle size, oocyte recovery rate on aspiration, and oocyte morphology in the mare

Oocytes were collected by aspiration of follicles from horse ovaries obtained at surgery or post-mortem. The oocytes were classified according to morphology of the ooplasm and cumulus. The size of the corresponding follicles was measured, and sections of the follicles were fixed and examined histologically to determine the stage of viability or atresia. In Part 1, 11 pairs of ovaries were examined and all follicles were sectioned; in Part 2, 9 pairs of ovaries were examined and only those follicles from which oocytes were recovered were sectioned. The number of follicles examined per pair of ovaries in Part 1 (average +/- SD) was 12.9 +/- 4.1. The proportion of follicles that were viable increased with increasing follicular size (P < 0.01); the percentage of viable follicles was 21, 42 and 83% for follicles < 10 mm, 10 to 19 mm, and >/= 20 mm in diameter, respectively. The overall oocyte recovery rate on aspiration of follicles was 46%. There was no significant difference in the oocyte recovery rate between viable and atretic follicles. A significantly higher proportion of oocytes recovered from viable follicles had granular ooplasm (64 vs 39%; (P < 0.05); whereas significantly more oocytes from atretic follicles had a misshapen or dense ooplasm (23 vs 6%; P < 0.05), or an expanded or pyknotic cumulus (24 vs 6%; P < 0.05). The most common cumulus morphology (63% of oocytes from viable follicles and 48% of oocytes from atretic follicles) was presence of only the corona radiata. Only 11% of oocytes from viable follicles and 9% of oocytes from atretic follicles had a complete cumulus present.     

Factors affecting follicular population, oocyte yield and quality in camels (Camelus dromedarius) ovary with special reference to maturation time in vitro

Three experiments were conducted to study a series of factors affecting in vitro reproductive parameters in camels. In Experiment 1, the effect of season and presence of a corpus luteum (CL) on ovarian follicular populations, oocyte yield and quality was studied using a total of 252 and 208 ovaries collected during the breeding and non-breeding season, respectively. Small, medium, large and the total number of ovarian follicles, oocyte yield and quality were measured. In Experiment 2, the effect of methods of oocyte retrieval and needle gauge on oocyte yield and quality was evaluated with oocytes recovered using slicing and aspiration with 18-, 19- or 20-gauge needle. Oocytes were evaluated microscopically and classified into three categories. The objective of Experiment 3 was to identify the optimum time for oocyte maturation in the dromedary camel. Oocytes were cultured in CR1aa medium at 38.5 degrees C under 5% CO(2) for 24, 32, 36, 48 and 72h. Maturation was calculated as the percentage of cumulus expansion and oocytes reaching metaphase II (MII). The number of small, medium, large and the total number of ovarian follicles were higher (P<0.01) during the breeding than non-breeding season. The recovery of total number of oocytes and Category I oocytes were also greater (P<0.01) during the breeding season. Ovaries without a CL possessed significantly (P<0.01) more ovarian follicles and more (P<0.05) small and large follicles. The total number of oocytes and Category I oocytes were also greater (P<0.01) in ovaries without CL. Slicing of camel ovaries increased (P<0.01) the yield of oocytes as compared to aspiration. The aspiration of follicles using a 20-gauge needle had greater yields of the total number of oocytes and Category I oocytes than when using 19- (P<0.05) and 18-gauge needle (P<0.01). The culture of camel oocytes for 36h produced higher (P<0.01) percentages of cumulus expansion and oocytes at MII. Increasing culture times up to 48 or 72h increased (P<0.01) the percentage of degenerated oocytes.In conclusion, the growth and development of ovarian follicles in the camel as well as yields of Category I oocyte were greater during the breeding season. Slicing or aspirations using a 20-gauge needle yielded greater numbers of total and Category I oocytes. Finally, maturation of oocytes in CR1aa medium for 36h produced higher percentages of cumulus expansion and oocytes at MII stage.     

Changes in ovarian, follicular, and oocyte morphology immediately after the onset of puberty are not accompanied by an increase in oocyte developmental competence in the pig

This study was conducted to determine whether ovarian morphology and developmental competence of in vitro-matured (IVM) oocytes is immediately affected by the onset of puberty in the pig. Ovaries of peri-pubertal pigs were sorted into two groups according to the presence or absence of corpora lutea presence (CL and NCL, respectively. Ovary dimensions, follicle diameter and number, and oocyte diameter (with and without zona pellucidae) were determined. The developmental competence of in vitro-matured oocytes from these two groups was evaluated following parthenogenetic activation and culture in vitro. CL ovaries were significantly (P<0.01) larger than NCL ovaries (width: 22.3+/-0.9 mm versus 15.9+/-0.4 mm, length: 33.2+/-1 mm versus 24.1+/-0.4 mm). Although CL ovaries had fewer antral follicles in total compared with NCL ovaries (21.1+/-1.8 mm versus 46.8+/-2.2 mm), they had a similar number of follicles 3-8mm in diameter. The mean diameter of follicles that were aspirated was greater for CL ovaries than for NCL ovaries (4.5+/-0.1 mm versus 3.3+/-0.02 mm). Oocytes from CL ovaries were greater in diameter compared with those from NCL ovaries (zona retained: 159+/-1.3 microm versus 146.1+/-1.5 microm, zona free: 124.7+/-1.8 microm versus 113.1+/-1.6 microm). No differences were found between oocytes from CL and NCL ovaries for rates of meiotic maturation (91.6+/-3.2% versus 92.4+/-3.2%), cleavage (88.4+/-11% versus 90.7+/-2.6%) and blastocyst formation (21.0+/-3.7% versus 23.7+/-5.7%). Therefore, the onset of puberty coincides with immediate changes in ovarian morphology, increased ovary size, follicle and oocyte diameter, but not with improved oocyte developmental competence. This suggests that the higher developmental competence usually observed in adult oocytes is acquired gradually and requires exposure to multiple estrus cycles.     

Animal and temporal effects on ovarian follicular dynamics in Brahman heifers

The aims of the current study were to determine if the pattern of ovarian follicular growth and development in Bos indicus heifers is different to that reported in Bos taurus breeds, and to examine the factors that determine which dominant follicle will ovulate. In addition, the extent to which variation in follicular dynamics is attributable to variation between animals and over time was evaluated. The ovaries of 17 Brahman heifers were examined daily by transrectal ultrasonography using a 7.5 MHz transducer for a total of 117 interovulatory intervals over a period of 10 months. Size and position of individual follicles ⪖5 mm in diameter, and size of corpora lutea (CL) were recorded. Circulating progesterone concentrations were determined from plasma samples obtained twice weekly. Although size of dominant follicles and CL within the ovaries of Bos indicus heifers were smaller than reported for Bos taurus breeds, the overall patterns of dominant follicle growth were similar. There were significant correlations between number of dominant follicles occurring prior to ovulation and time of appearance of the second dominant follicle, duration of detection of CL and size of the ovulatory follicle in the preceding oestrous cycle (P < 0.05). There were significant animal effects on a number of ovarian characteristics including number of dominant follicles per oestrous cycle (P < 0.001), with one heifer having four dominant follicles in more than a third of oestrous cycles observed. In addition, changes in daylength over the 10 month period were related to changes in duration of the interovulatory interval, persistence and maximum diameter of CL and size of ovulatory follicles. Liveweight change over the same period was related to changes in maximum diameter of the first dominant follicle.     

Morphological changes of the ovary and hormonal changes through the ovarian cycle of the common marmoset (Callithrix jacchus)

Laparoscopic observations of morphological changes of the ovary during the ovarian cycle in conjunction with radioimmunoassay of serum progesterone and estradiol-17β was investigated as a method of monitoring the ovarian cycle in the common marmoset. In the common marmoset, plural follicles first appeared in each ovary five days prior to ovulation. At three to four days prior to ovulation one or two follicles developed into translucent blisters on the surface of the ovary. As the follicles filled with follicular fluid, they became larger and clearer until one to two days prior to ovulation, at which time they formed well defined, transparent bubbles protruding from the surface of the ovary. After ovulation, the ovulation point could be detected at the center of the follicle, sometimes surrounded by a corpus of engorged blood vessels. Ovulations of the plural follicles were not simultaneous, and due to the time lag ovulations took at least 12 to 20 hrs in four out of seven animals examined. After two to five days of ovulation the corpus hemorragicum, a bright red protrusion made of tissue and blood disrupted by ovulation, was found. Subsequently, the color of the formatted corpus luteum changed from dark-red to yellow then to yellow white. While the corpus luteum remained reddish in color serum progesterone was maintained at as high levels as in the luteal phase. There was no mature follicle or corpus luteum in subordinate female ovaries.     

成体雌性小熊猫卵巢组织学观察

2000 年至2009 年,12 只固定于10% 福尔马林中非生殖系统疾病死亡的小熊猫卵巢组织,按常规组织学技术制作组织切片,HE 染色,光学显微镜观察。结果:(1)不同发情时期卵巢均有原始卵泡、初级卵泡和次级卵泡分布。发情期的卵巢未观察到典型的成熟卵泡和卵母细胞; (2)原始卵泡数量较少,初级卵泡数量较多,多数初级卵泡和大多数的次级卵泡都处在闭锁状态;(3)卵泡腔出现之前,卵母细胞的直径和卵泡直径同时增长;卵泡腔出现之后,卵母细胞直径增长较慢,卵泡直径增长较快; (4)不同发情时期的小熊猫卵巢均存在大量的间质腺细胞;(5)妊娠小熊猫和发情间期无妊娠小熊猫的卵巢均有发育正常的黄体;(6)卵泡细胞发育呈低柱状至柱状时出现透明带。结论:(1) 卵泡闭锁主要发生在初级卵泡阶段,仅少数卵泡能发育至次级卵泡;(2)卵母细胞和卵泡生长呈双相生长的趋势; (3) 不同发情时期的小熊猫卵巢间质腺都发达; (4)发情排卵后,非妊娠黄体与妊娠黄体维持的时间相似,证实了小熊猫存在假孕现象。     

Ultrasonographic study of the effects of the corpus luteum on antral follicular development in unilaterally ovulating western white-faced ewes

The objective of this study was to examine the local effects of the corpus luteum (CL) on ovarian antral follicle development by looking at follicle populations and dynamics in ovaries with or without CL, in unilaterally ovulating ewes, using a retrospective analysis of daily ultrasonographic records. The present report summarises the data from the first luteal phase of the breeding season (August-October; n = 4), a luteal phase in the mid-breeding season (November-December; n = 5), the last luteal phase of the breeding season (January-March; n = 5), and the luteal phase after GnRH-induced ovulations in mid-anoestrus (May-June; n = 4) of western white-faced ewes. Mean daily numbers of 3mm follicles that did not grow any larger were significantly reduced in the CL-containing ovaries of ewes at all periods of study except for the transition to anoestrus. With all scanning periods combined, daily numbers of 3mm follicles not growing further increased (P<0.05) between day 6 and 15 after ovulation in the CL-containing ovaries. Based on mean data for the whole periods of observation, the non-CL-bearing ovaries of ewes in the transition to anoestrus had fewer (P<0.05) follicles growing from 3 to > or =5mm in size before regression compared with the mid-breeding season and mid-anoestrus. The lifespan of follicles reaching > or =5mm in diameter was shorter (P < 0.05) in the CL- compared with non-CL-containing ovaries of anoestrous ewes induced to ovulate with GnRH ((6.5+/- 1.3) and (9.0+/- 1.0) days, respectively). Circulating concentrations of progesterone were lower during both transitional periods (into and out of anoestrus) and mid-anoestrus than during the mid-breeding season (P < 0.001), and were less during anoestrus than during both transitional periods (P < 0.05). It was concluded that CL/luteal structures locally suppressed the growth of ovarian antral follicles to the 3mm size-range except during the transition to anoestrus, but that there was no inhibitory effect of the CL on the growth of ovarian follicles to larger diameters. The presence of CL/luteal structures did not affect the length of the lifespan of follicles reaching > or =5mm in diameter nor the number of ovulations per ovary in cyclic ewes, but shortened large follicle lifespan in anoestrous ewes. Variations in peripheral concentrations of progesterone across the breeding season and between the breeding season and anoestrus did not alter the lifespan of large antral follicles. In the transition to anoestrus and during mid-anoestrus, the presence of the CL in an ovary appeared to maintain follicle development to ovulatory sizes and to increase the rate of turnover of large antral follicles, respectively.     

Systemic and intraovarian effects of corpus luteum on follicular dynamics during estrous cycle in hair breed sheep

The present study aimed to determine systemic and local effects of corpora lutea (CL), on follicular dynamics throughout the estrous cycle. All follicles >or=2 mm and CL were assessed by daily transrectal ultrasonography in 12 West African ewes. Blood samples were collected to determine plasma concentration of progesterone. Fifteen estrous cycles were evaluated with a mean interovulatory interval of 16.8+/-0.2 days. Two (13.3%), 10 (66.7%) and 3 (20%) of the estrous cycles had 2, 3 and 4 waves of follicular development, respectively. In sheep with three waves of follicular development, both the length of growing phase and the growth rate of dominant follicles from midluteal wave II were diminished (3.4+/-0.3 days, P<0.0001, and 0.4+/-0.1 mm/day, P<0.01, respectively) when compared to follicles from early luteal phase (wave I, 4.1+/-0.2 days, and 0.7+/-0.1 mm/day) or late luteal phase (wave III, 6.3+/-0.4 mm and 0.6+/-0.1 mm/day). The diameter of the dominant follicle was smaller during the midluteal phase (3.9+/-0.1 mm, P<0.0001) than in the early and late luteal phase (5.0+/-0.2 and 5.7+/-0.2 mm; respectively). The effect of the dominant follicle was less during midluteal phase, because number of accompanying smaller follicles was fewer (P<0.01) in waves I and III (6.3+/-0.9 compared with 3.4+/-0.8 and 2.3+/-0.7). The number of follicles was also different between ovaries that had CL and those that did not. The total number of large follicles during the luteal phase was less in ovaries with CL (0.9+/-0.5 compared with 2.7+/-0.3; P<0.01), as was the mean daily number of both large (0.1+/-0.02 compared with 0.2+/-0.02; P<0.001) and total number of follicles >or=2 mm (2.5+/-0.1 compared with 3.3+/-0.1; P<0.01). Current results indicate that the presence of a functional CL may exert both systemic and local effects on the population of follicles, affecting the dominance exerted by large follicles.     

Follicular dynamics and plasma FSH and progesterone concentrations during follicular deviation in the first post-ovulatory wave in Nelore (Bos indicus) heifers

The objective of the present study was to characterize ovarian follicular dynamics and hormone concentrations during follicular deviation in the first wave after ovulation in Nelore (Bos indicus) heifers. Ultrasonographic exams were performed and blood samples were collected every 12h from the day of estrus until 120-144 h after ovulation in seven females. Deviation was defined as the point at which the growth rate of the dominant follicle became greater than the growth rate of the largest subordinate follicle. Deviation occurred approximately 65 h after ovulation. Growth rate of the dominant follicle increased (P<0.05) after deviation, while growth rate of the subordinate follicle decreased (P<0.05). Diameter of the dominant follicle did not differ from the subordinate follicle at deviation (approximately 5.4mm). The dominant follicle (7.6mm) was larger (P<0.05) than the subordinate follicle (5.3mm) 96 h after ovulation or 24h after deviation. Plasma FSH concentrations did not change significantly during the post-ovulatory period. The first significant increase in mean plasma progesterone concentration occurred on the day of follicular deviation. In conclusion, the interval from ovulation to follicular deviation (2.7 days) was similar to that previously reported in B. taurus females, but follicles were smaller. Diameters of the dominant follicle and subordinate follicle did not differ before deviation and deviation was characterized by an increase in dominant follicle and decrease in subordinate follicle growth rate. Variations in FSH concentrations within 12-h intervals were not involved in follicular deviation in Nelore heifers.