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Immediate and delayed effects of heat stress on follicular development and its association with plasma FSH and inhibin concentration in cows.

The aim of this study was to characterize the immediate effects of heat stress on plasma FSH and inhibin concentrations, and its involvement in follicular dynamics during a complete oestrous cycle, and to examine a possible delayed effect of heat stress on follicular development. Holstein dairy cows were oestrous synchronized and randomly assigned to either cooled (n = 7) or heat-stressed (n = 6) treatment groups. During a complete oestrous cycle, control cows, which were cooled, maintained normothermia, whereas heat-stressed cows, which were exposed to direct solar radiation, developed hyperthermia. At the end of this oestrous cycle (treated cycle), both groups were cooled and maintained normothermia for the first 10 days of the subsequent oestrous cycle. Throughout this period, follicular development was examined by ultrasonography, and plasma samples were collected. During the second follicular wave of the treated oestrous cycle, a significantly larger cohort of medium sized follicles (6-9 mm) was found in heat-stressed cows than in cooled cows (P < 0.05). The enhanced growth of follicles in this wave in heat-stressed cows was associated with a higher plasma FSH increase which lasted 4 more days (days 8-13 of the oestrous cycle; P < 0.05), and coincided with a decrease in the plasma concentration of immunoreactive inhibin (days 5-18 of the oestrous cycle; P < 0.05). During the follicular phase (days 17-20 of the treated cycle), heat-stressed cows showed an increase in the number of large follicles (>/= 10 mm), and the preovulatory plasma FSH surge was significantly higher in heat-stressed cows than in cooled cows (P < 0.01). The effect of heat stress was also observed during the first follicular wave of the subsequent cycle: the postovulatory plasma FSH concentration was higher (P < 0.01), but fewer medium follicles developed, and the first follicular wave decreased at a slower rate in previously heat-stressed cows than in cooled cows (0.40 and 0.71 follicles per day, respectively). This study shows both immediate and delayed effects of heat stress on follicular dynamics, which were associated with high FSH and low inhibin concentrations in plasma. These alterations may have physiological significance that could be associated with low fertility of cattle during the summer and autumn.

Animals↗

The effect of feeding strategy during the pre-follicular phase on subsequent follicular development in the pig.

In female pigs feeding level has important effects on reproductive performance. This review is focused on the follicular development after low and high feeding levels during the luteal phase in gilts. Although aspects of diet composition seem to have a role in regulating reproductive performance, the most important aspect appears to be the plane of nutrition. Similar effects are described during lactation in primiparous sows, when their metabolism is challenged to its maximum. Also in this situation feeding level clearly affects weaning to oestrus interval, follicular development, oocyte maturation, ovulation rate and subsequent embryonic survival.

Animal Nutritional Physiological Phenomena↗

Characteristics of gonadotropin response, follicular development, and endometrial growth and maturation across consecutive cycles of clomiphene citrate treatment.

OBJECTIVE: To examine the patterns of gonadotropin response, follicular development, and endometrial growth and maturation across consecutive cycles of clomiphene citrate (CC) treatment. DESIGN: Prospective analysis of cycle characteristics. SETTING: Academic tertiary medical center. PATIENTS: Nineteen consenting anovulatory infertile women receiving standardized, cyclic, incremental treatment with CC (50 to 150 mg/d, cycle days 5 to 9) for ovulation induction. INTERVENTIONS: In each of up to six consecutive treatment cycles, urinary LH was monitored twice daily from cycle day 10 until detection of the LH surge or day 21; blood samples and transvaginal ultrasound (US) examination were obtained on cycle days 3, 10, and every 1 to 3 days thereafter until collapse of the dominant follicle. Endometrial biopsy was performed 11 to 13 days after the LH surge in the first, third, and sixth ovulatory cycle. RESULTS: Follicular phase duration, peak follicular diameter, the number of preovulatory follicles, and peak endometrial thickness and echo pattern remained consistent across consecutive ovulatory (n = 55) and anovulatory (n = 23) treatment cycles. Endometrial dating was > or = 3 days out of phase in 2 of 31 (6%) cycles sampled. Peak serum E2 and P concentrations did not vary with cycle number or correlate with endometrial thickness or echo pattern. Cycle day 10 FSH levels were significantly higher in ovulatory subjects than in anovulatory subjects. CONCLUSIONS: Patterns of gonadotropin response, follicular development, and endometrial growth and maturation remain consistent across consecutive cycles of CC treatment.

Adult↗

Expression and function of Fas antigen vary in bovine granulosa and theca cells during ovarian follicular development and atresia.

Fas antigen is a receptor that triggers apoptosis when bound by Fas ligand (FasL). A role for Fas antigen in follicular atresia was studied in follicles obtained during the first wave of follicular development during the bovine estrous cycle (estrus is Day 0). Granulosa and theca cells were isolated from healthy dominant follicles and the two largest atretic subordinate follicles on Day 5, atretic dominant follicles on Days 10-12, and preovulatory follicles on Day 1. Fas antigen mRNA levels were highest in granulosa cells from subordinate as compared to other follicles, and lowest in theca cells from healthy Day 5 dominant as compared to other follicles. FasL alone had no effect on viability of granulosa or theca cells but became cytotoxic in the presence of interferon-gamma (IFN). IFN has been shown to induce responsiveness to Fas antigen-mediated apoptosis in other cell types. In the presence of IFN, killing of granulosa cells by FasL was greater in subordinate compared to healthy dominant follicles on Day 5, did not differ between healthy and atretic dominant follicles, and was similar in theca among all follicles. Granulosa cells from preovulatory follicles, which had been exposed to the LH surge in vivo, were completely resistant to FasL-induced killing. In summary, Fas antigen expression, and responsiveness to Fas antigen-mediated apoptosis, vary during follicular development.

Animals↗

Follicular development during the prepuberal period of different morphological types of ovaries in Hampshire and Yorkshire gilts fed two planes of nutrition.

Changes in follicular development of the ovary at 105, 140 and 175 d of age were observed on 48 prepuberal Yorkshire and Hampshire gilts raised on two planes of nutrition. The follicles from the left ovary were counted and measured by histological techniques. Both the nonatretic and the atretic (more than four pyknotic bodies) antral follicles were classified into six categories according to size, and each category was expressed as percentage of the total number. The proportion of nonatretic follicles belonging to the first category (.19 to .36 mm in diameter) was higher in Hampshire than Yorkshire gilts at d 105 (65.4 vs 47.6%; P less than .05), but not at d 140 and 175. The proportion of atretic follicles to the total number observed was higher in Yorkshire at d 175 than at d 140 and 105 (38.6 vs 21.9 vs 11.6%, respectively; P less than .05), whereas in Hampshire, significant differences were observed only between d 140 and 105 (25.2 vs 3.9%, respectively). The low-plane regimen reduced the percentage of nonatretic follicles of the third (.63 to 1.12 mm) and fourth (1.13 to 2.00 mm) categories in Yorkshire gilts from 14.4 and 7.2% to 9.7 and 4.0%, respectively (P less than .05). According to grouping, size and number of antral follicles, three morphological types of ovaries could be distinguished: honey-comb, grape-like and an intermediate class. Reanalyzing the data accounting for types resulted in radical reduction in the residual variation, hence many significant differences between the two breeds could be identified. At 140 and 175 d of age, Hampshires with honey-comb- and grape-like ovaries had greater numbers of medium and large follicles than Yorkshires (P less than .001). It is concluded that the slower follicular development observed at 105 d of age in Hampshire, as compared with Yorkshire gilts, allows accumulation of greater numbers of antral follicles at 140 and 175 d for both morphological types of ovaries.

Animals↗

Effects of exogenous steroids on serum FSH and LH, and on follicular development in cyclic mares.

Cyclic mares were given daily i.m. injections of 150 mg progesterone (Group P, N = 4), 150 mg progesterone and 10 mg oestradiol-17 beta (Group PE, N = 3), 10 mg oestradiol-17 beta (Group E, N = 3) or cottonseed oil vehicle (Group C, N = 4), from the day after ovulation (Day 1) to Day 28. Blood samples were collected daily, and the ovaries were palpated every 1-2 days. Serum FSH and LH concentrations were measured in all samples, and means determined for 7 consecutive 4-day periods throughout treatment. Comparisons within each steroid treatment group between time periods and comparisons between hormone treatment groups within each time period were made to investigate the way in which these ovarian steroids control cyclic gonadotrophin changes in the mare. The increase in LH during oestrus in Group C mares (controls) appeared to be inhibited by progesterone, resulting in low LH concentrations and failure of preovulatory-sized follicles to ovulate. In Group PE LH concentrations were lower than those in Group P, resulting in suppression of the development of the largest follicle. In Group E, treatment had little effect on FSH and LH concentrations, while follicular development was variable (ovulations on Days 25, 33 and 36). None of the steroid treatments appeared to suppress FSH concentrations directly but FSH concentrations showed a reciprocal relationship with the LH-dependent follicular development.

Animals↗

Involvement of the Fas/Fas ligand system in p53-mediated granulosa cell apoptosis during follicular development and atresia.

In the present study we have examined the presence of Fas, Fas ligand (FasL), and p53 in rat granulosa cells during follicular development and atresia, especially in relation to the granulosa cell cycle progression and the onset of granulosa cell apoptosis. Fas, FasL, and p53 proteins were immunolocalized, and their contents were determined by Western blotting. Granulosa cell apoptosis was assessed by DNA fragmentation analyses (DNA ladder) and in situ terminal deoxynucleotidyl transferase mediated deoxy-UTP-biotin nick end labeling (TUNEL) as well as by flow cytometry. Ovaries not exposed to gonadotropins (control) consisted predominantly of preantral and early (small) antral follicles, the latter of which were mostly atretic and demonstrated intense TUNEL staining in granulosa cells exhibiting positive immunoreactivities for FasL and Fas. Granulosa cells isolated from these follicles were apoptotic, as evident by clear ladder pattern of DNA fragmentation upon electrophoretic analysis and the high percentage (>10%) of the cell population in the A0 phase of the cell cycle. After gonadotropin treatment, these features completely disappeared during each of the 3 days of follicular growth to the medium to large antral stages. Cell cycle analysis showed significantly higher proportion of the cells in S and G2/M phases compared with controls, which was accompanied by marked decrease in immunoreactivities for Fas, FasL, and p53. By days 4 and 5, widespread atresia and extensive granulosa cell apoptosis were noted in large antral and preovulatory follicles and were coincidental to increased expression of p53 and Fas, but not of FasL, as well as an apparent arrest of granulosa cell G1/S progression, as evident by an increased cell population in G0/G1 and a decrease in the S and G2/M. Granulosa cells from equine CG-primed ovaries exhibited marked increases in p53 and Fas protein contents and apoptosis after adenoviral p53-sense complementary DNA infection in vitro and were more responsive to Fas activation by an agonistic Fas monoclonal antibody challenge. Taken together, these findings are consistent with the well accepted concept that gonadotropin plays a central role as a survival factor in the regulation of granulosa cell Fas/FasL and p53 expression during ovarian follicular development. In addition, the control of granulosa cell apoptosis may involve two consecutive cellular/molecular events: cell cycle arrest at G1/S and exit from G0 into A0 phase, via regulation of the p53 and Fas/FasL death pathways.

Animals↗

Requirements for human chorionic gonadotropin and recombinant human luteinizing hormone for follicular development and maturation.

PURPOSE: Our purpose was to evaluate the requirements for human chorionic gonadotropin (hCG) and recombinant luteinizing hormone (rec-LH) for follicular development and maturation in mice. METHODS: We carried out ovarian stimulation of immature mice. Output parameters were the preembryos created in vivo and frequency of blastocyst formation in vitro. RESULTS: hCG at 0 to 1 IU resulted in a dose-dependent recovery of preembryos (0 to 39.7 +/- 4.3; mean +/- SE) per mouse. hCG at 1 and 10 hCG gave similar results, whereas higher doses significantly reduced the number of preembryos. Potential for blastocyst formation was independent of hCG dose. hCG and rec-LH together exerted a synergistic effect on the recovery of preembryos. CONCLUSIONS: Optimal follicular development required a combination of 20 IU follicle stimulating hormone and 1-10 IU hCG. The potency of hCG was higher than that of rec-LH, but a synergistic effect of rec-LH and hCG was observed. The results may be pertinent for the development of strategies for ovarian stimulation of women with low levels of endogenous LH.

Animals↗

Ovarian follicular development in the rat: hormone receptor regulation by estradiol, follicle stimulating hormone and luteinizing hormone.

The effects of estradiol, FSH and LH on ovarian follicular development and granulosa cell differentiation were examined in the immature rat hypophysectomized on day 24 of age. Administration of estradiol to hypophysectomized rats for 4 days stimulated the growth of large preantral follicles with a concomitant 1.5-fold increase in FSH receptor content and a 4-fold decrease in LH receptor content in the granulosa cells. When highly purified hFSH was administered alone, receptor content for FSH increased progressively for 4 days while receptor for LH remained essentially unchanged. However, when rats were pretreated with estradiol, the response of follicles to FSH was markedly enhanced as indicated by the appearance of large, antral follicles and elevated receptor content for both FSH and LH. Receptor content for FSH increased markedly in response to hFSH following only one day of estradiol pretreatment, while receptor content for LH increased most rapidly in response to hFSH after 3 days of estradiol pretreatment. LH administered to rats possessing large preovulatory follicles caused luteinization of granulosa cells and a marked decline in receptor content for both gonadotropins within 24 h. Receptor content remained low even 48 h after LH administration when granulosa cells were fully luteinized. These results indicated that follicular development and granulosa cell differentiation are dependent on steroid-protein hormone regulation of hormone specific receptors.

Animals↗

Effect of multiple follicular development on uterine perfusion in subsequent spontaneous cycles.

OBJECTIVE: To investigate the effect of ovarian hyperstimulation on uterine perfusion during a subsequent natural cycle to evaluate if there is a long-term effect of treatment with gonadotropins. STUDY DESIGN: Thirty-five infertile couples were subjected to three cycles of multiple follicular development. Uterine blood flow was determined during the follicular phase, ovulation and luteal phase of the spontaneous cycle preceding and the one following the stimulated cycles. RESULTS: No significant difference was present among pulsatility index (PI) values in the two cycles. PI values (+/- SD) were 2.05 +/- 0.5, 2.28 +/- 0.4, 2.18 +/- 0.5, 2.13 +/- 0.4 and 2.00 +/- 0.5, respectively, in the pretreatment cycle and 2.32 +/- 0.5, 2.56 +/- 0.6, 2.42 +/- 0.3, 2.37 +/- 0.4 and 2.29 +/- 0.5, respectively, in the posttreatment cycle. An inverse correlation was found between PI values and estradiol in the follicular phase of the first spontaneous cycle. As for radial and subendometrial arteries, no significant difference was present for PI values in the two cycles. CONCLUSION: Multiple follicular development seems not to have a long-term effect on uterine perfusion in the subsequent spontaneous cycle.

Adult↗

Effect of estrogen administration on endogenous and luteinizing hormone-releasing-hormone-induced luteinizing hormone secretion and follicular development in the lactating sow.

The objectives of this study were to investigate whether estradiol treatment during lactation modifies 1) the patterns of endogenous LH, FSH, and prolactin (PRL) release; 2) the sensitivity of the pituitary to exogenous injections of LHRH; and 3) the responsiveness of the ovarian follicles to gonadotropin. Plasma LH, FSH, and PRL were determined in samples taken repeatedly from 18 sows on Days 24-27 of lactation. Ovaries were then recovered, and follicular development was assessed by measuring the follicular diameter (FFD) and follicular fluid estradiol-17 beta concentration (FFE) of the ten largest follicles dissected from each ovary. Sows were randomly allocated to one of four treatments: 1) Group C (4 sows) received no treatment; 2) Group LHRH (5 sows) received 800 ng of LHRH every 2 h throughout the sampling period; 3) Group E2 (4 sows) received subcutaneous implants containing estradiol-17 beta 24 h after start of sampling; 4) Group LHRH + E2 (5 sows) were administered a combination of LHRH and estradiol-17 beta implants. Between-animal variability for plasma LH, FSH, and PRL was considerable. LH concentration and LH pulse frequency increased (p less than 0.05) after LHRH treatment in the LHRH and LHRH + E2 groups; however, an acute inhibition of LH secretion was observed in the latter group immediately after estradiol implant application. In the absence of LHRH treatment, estradiol caused chronic inhibition of LH secretion. Follicular development was greater in the LHRH and LHRH + E2 groups compared to the C and E2 groups (p less than 0.05 for both FFD and FFE).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Follicular development and steroid concentrations in cows with different levels of fertility raised under nutritional stress.

The aim of the present study was to characterize ovarian follicular development and steroid concentrations during postpartum and the estrous cycle of Brangus Ibagé cows (3/8 Nelore + 5/8 Aberdeen Angus) with different levels of fertility. Cows were classified as having high or low fertility according to the calving interval (CI). The average CI of the herd from which cows used in this study were selected was 404.6+/-5.44 and 711.2+/-20.89 days for the high and low fertility groups, respectively. Four cows of high fertility and five cows of low fertility had calves removed between 70 and 100 days after parturition. Ovarian activity was monitored daily by ultrasound for 16 days after calf removal. Days to emergency of the first follicular wave after calf removal, number of follicles with diameter >9 mm, growth rate of largest follicle, maximum diameter of largest follicle, length (days) and number of follicular waves were recorded. During this period, blood was collected daily for measurements of serum progesterone (P(4)) and estradiol (E(2)) concentrations. In another experiment, ovarian activity and P(4) and E(2) concentrations were examined during estrous cycle in five cows of high fertility and four cows of low fertility. Ovarian activity and steroid concentrations were assessed from the day prior to estrus to the 15th day of the estrous cycle (estrus = day 0). In postpartum cows of high fertility, the total number of follicles >5mm and the maximum diameter of the largest follicle were higher than in cows of low fertility (P < 0.05). Concentrations of P(4) and E(2) did not differ between groups in the postpartum cows. However, E(2) increased 5 days after calf removal (around 90 days of postpartum) in the high fertility group, followed by an increase in P(4) with average values indicating ovulation around 100 days postpartum. In cycling cows, the profile of follicular development was similar between cows of high and low fertility. There was no difference between groups for number of follicles >5mm, but the day effect was significant (P < 0.01). Plasma concentrations of P(4) and E(2) were similar in both groups. These data suggest that cows, from a population raised in the same environment have different fertility as a consequence of individual physiological characteristics.

Animal Nutritional Physiological Phenomena↗

Connexin 43 gap junction protein expression during follicular development in the porcine ovary.

Connexin 43, a member of the highly conserved connexin family of gap junction proteins, is expressed in the pig ovary. In other species, ovarian connexin 43 expression and phosphorylation are hormonally regulated. We characterized connexin 43 expression and phosphorylation in the ovaries of mature pigs during the estrous cycle and in prepubertal gilts during follicular development induced by eCG (750 IU)/hCG (500 IU; 72 h later). Ovarian connexin 43 protein expression and phosphorylation were examined by immunoblot analysis. Connexin 43 was localized to specific follicular cell types during development by immunofluorescence. While no change in total connexin 43 protein expression was seen during the cycle, connexin 43 phosphorylation was significantly higher (p < 0.05) during the late follicular stage of the cycle than during the early luteal and early to mid-follicular stages. In ovaries of eCG/hCG-primed prepubertal pigs, connexin 43 protein levels remained steady, while phosphorylation of the protein increased significantly at 72 h and 84 h after eCG treatment (p < 0.05), then declined to pretreatment levels by 96 h (24 h post-hCG administration). Immunoreactive connexin 43 was localized predominantly to granulosa cells of cyclic pigs and eCG/hCG-primed prepubertal gilts. Follicular connexin 43 was highest between 60 h and 84 h after eCG and declined after hCG administration. Connexin 43 was not detected in morphologically atretic follicles, stroma, or vascular tissue of the ovary. This is the first evidence that porcine ovarian connexin 43 phosphorylation is differentially regulated during follicular development. The results suggest that hormonally induced changes in connexin 43 phosphorylation may play a coordinating role in porcine follicular development.

Animals↗

Human recombinant activin-A alters pituitary luteinizing hormone and follicle-stimulating hormone secretion, follicular development, and steroidogenesis, during the menstrual cycle in rhesus monkeys.

Activin, a stimulator of pituitary FSH secretion in nonprimate species, may also act in the ovary to modulate follicular development. To examine whether activin has similar actions in primates, female rhesus monkeys (n = 3/treatment) exhibiting regular menstrual cycles received sc injections of either vehicle or 60 micrograms/kg recombinant human activin-A at 0800 and 1600 h for 1 (acute) or 7 (chronic) days beginning in the early follicular phase. The vehicle-treated monkeys displayed menstrual cycles of normal length, with the follicular (11.3 +/- 1.3 days, mean +/- SE) and luteal (16.6 +/- 1.8 days) phases demarcated by midcycle peaks in serum estradiol (E) and bioactive LH. After the first activin injection, levels of human activin A peaked at 90 ng/mL within 1 h and returned to baseline before the second injection 8 h later. Although serum E and FSH levels did not change, LH increased (273%, P < 0.05) within 8 h. Acute activin treatment increased (P < 0.05) serum E within 24 h to levels (1290 +/- 330 pmol/L) typically observed at midcycle. With chronic treatment, serum E peaked on day 2 (2580 +/- 338 pmol/L; P < 0.05), then declined and rose to a second peak (1680 +/- 279 pmol/L) on day 5. During chronic activin treatment, LH levels peaked on day 2 (603 +/- 270 ng/mL; P < 0.05 compared to day 0, 15 +/- 7 ng/mL) whereas FSH increased progressively until day 5 (937 +/- 320 ng/mL; P < 0.05 compared to day 0, 169 +/- 59 ng/mL). After acute or chronic activin, the expected midcycle rises in serum E and gonadotropins were delayed to greater than or equal to day 20 (n = 4) or did not occur before menses (n = 2). Although an enlarged ovary with one greater than or equal to 4-mm follicle was observed by laparoscopy during the late follicular phase in vehicle-treated monkeys, medium-to-large follicles were not visible on ovaries during chronic activin treatment or later at the expected midcycle interval in activin-treated monkeys. Similar hormonal and ovarian events were obtained after activin treatment of amenorrheic monkeys having serum FSH, LH, and E levels that were comparable to those at menses in spontaneous menstrual cycles. Thus, exogenous activin stimulates pituitary LH and FSH secretion and ovarian estrogen secretion during the early follicular phase in intact monkeys. However, acute or chronic activin treatment did not promote complete follicular development and disrupted subsequent events in the menstrual cycle.(ABSTRACT TRUNCATED AT 400 WORDS)

Activins↗

Effects of oxytocin on follicular development and duration of the estrous cycle in heifers.

Holstein heifers were used to study effects of exogenous administration of oxytocin on luteal function and ovarian follicular development. Twelve heifers were monitored for 1 estrous cycle to confirm normal ovarian function. At the subsequent estrus, these animals were randomly assigned to 1 of 3 treatments: saline control, (Group 1, n=4), oxytocin (Group 2, n=4) and saline pregnant (Group 3, n=4). Group 2 received continuous infusion of oxytocin (1.9 mg/d) from Days 14 to 26 after estrus, while Groups 1 and 3 received saline infusion during the same period. Group 3 were artificially inseminated at estrus. Daily blood samples were collected for oxytocin and progesterone assay. Ovarian follicles and corpus luteum (CL) development were monitored daily by transrectal ultrasonography until Day 32 after estrus. Plasma progesterone (P4) concentrations prior to initiation of infusion were 7.6+/-1.3 ng/mL on Day 14. They then decreased to <1 ng/mL on Day 19 for Group 1 and on Day 28 for Group 2. The interestrous interval was longer (P <0.05) for heifers that received oxytocin infusion. During the infusion period P4 concentrations were not different (P >0.05) between Group 2 and 3 but declined gradually from Day 20 in Group 2 despite the presence of high plasma oxytocin concentrations. Control heifers had 2 waves of follicular growth, with the second dominant follicle ovulating. Three of the 4 oxytocin-infused animals had an additional wave, with the third dominant follicle ovulating. Oxytocin infusion had no effect on size of the ovulating follicle (P >0.05) and the number of Class 1 follicles (3 to 5 mm, P >0.1). Differences in the number of Class 2 follicles (6 to 9 mm) among treatments on Days 15 to 22 after estrus were not detected (P >0.1) except on Days 23 to 26, when Group 2 had fewer follicles than Group 3 (P <0.05). The results show that continuous infusion of oxytocin during normal luteolysis delays luteal regression without inhibiting follicular development.

Animals↗

Impact of dietary lysine intake during lactation on follicular development and oocyte maturation after weaning in primiparous sows.

Primiparous sows (n = 36) were used to evaluate the effects of dietary lysine intake in lactation on follicular development and oocyte maturation after weaning. Sows were assigned randomly to one of three diets containing .4% (low lysine, LL), 1.0% (medium lysine, ML), or 1.6% (high lysine, HL) total lysine. All diets contained 2.1 Mcal NE/kg and exceeded NRC (1988) requirements for all other nutrients. Actual lysine intakes over an 18-d lactation were 16, 36, and 56 g/d for sows consuming LL, ML, and HL, respectively. Ovarian data were analyzed for sows determined to have been slaughtered during the first proestrus period after weaning, using previously established criteria. Compared with sows fed ML and HL, sows fed LL tended to have lower uterine weight, follicular fluid volume, and follicular fluid (FF) estradiol (E2) content (P < .15), but similar ovarian weight and follicular fluid IGF-I concentration. Within the largest 15 preovulatory follicles, sows fed LL had a lower percentage of large (> or = 7.0 mm) follicles (33 vs 50 and 58%; P < .01) and a higher percentage of medium (5.5 to 7.0 mm) follicles (62 vs 44 and 39%; P < .01) but a similar percentage of small (< or = 5.5 mm) follicles (4.4 vs 5.9 and 3.7%; P > .15), respectively, compared with sows fed ML or HL. Standardized pools of oocytes aspirated from follicles of prepubertal gilts were incubated for 44 h with pooled FF recovered from the largest 15 follicles of each experimental sow. Fewer oocyte nuclei matured to metaphase II of meiosis when cultured with FF recovered from sows fed LL, than from sows fed ML or HL (47.1 vs 59.8 and 63.8%, respectively; P < .01). Our results suggest that low lysine (protein) intake in primiparous lactating sows impaired follicular development and reduced the ability of follicles to support oocyte maturation. However, high compared with medium lysine (protein) intake had no further positive effects on ovarian function.

Animal Nutritional Physiological Phenomena↗

Relationships between luteinizing hormone, follicle-stimulating hormone and prolactin secretion and ovarian follicular development in the weaned sow.

Folliculogenesis was studied by assessing development of the largest 10 follicles obtained from 10 sows 48 h after weaning and by analyzing changes in plasma luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL) for 24 h before weaning until 48 h after weaning. Follicular diameter, follicular fluid volume, and concentrations of estradiol and testosterone and granulosa cell numbers were determined in all follicles, and 125I-hCG binding to theca and granulosa and maximal aromatase activity in vitro was determined in five follicles/sow. Overall, a significant rise in LH, but not in FSH, occurred at weaning, although in individual sows an increase in LH was not necessarily related to subsequent estrogenic activity of follicles. In 9/10 sows, PRL fell precipitously after weaning. In lactation, LH was negatively, and after weaning, positively, correlated with FSH and PRL. Marked variability in follicular development existed within and between sows. Overall, most follicular characteristics were positively correlated to follicular diameter; however, in larger follicles the number of granulosa cells was variable and unrelated to estrogenic activity, which--together with theca and granulosa binding of hCG--increased abruptly at particular stages of follicular development. Differences in maturation of similarly sized follicles from different sows were related to estrogenic activity of the dominant follicles but not to consistent differences in LH, FSH or PRL secretion. Both the dynamics and the control of folliculogenesis in the sow, therefore, appear to be complex.

Animals↗

Some aspects of thecal and granulosa cell function during follicular development in the bovine ovary.

The patterns of ovarian follicular development and the steroidogenic properties of individual follicles (greater than or equal to 2 mm diam.) were assessed in Angus cows from Day - 5 until Day + 1 of the oestrous cycle (oestrus = Day 0). Individual follicles were judged to be healthy or atretic using a new classification system incorporating assessments of thecal vascularity and colour, the number of granulosa cells, the presence or absence of debris in follicular fluid and the status of the oocyte. The results suggest that the theca interna of small antral follicles (less than 5 mm diam.) responds to LH and synthesizes androstenedione before the granulosa cells develop an appreciable ability to metabolize androgen to oestrogen. Regardless of follicle size, the output of thecal androstenedione per unit mass of tissue remained unchanged in healthy but not in atretic follicles. On a per cell basis, aromatase activity increased in granulosa cells from healthy but not from atretic follicles with increasing follicle size. Peak levels of aromatizing activity were consistently observed in dominant oestrogen-enriched follicles on Day 0 although similar activity was also observed in some healthy follicles (greater than or equal to 8 mm diam.) on other days of the cycle. Early atresia in bovine follicles was characterized by an absence or lowering of aromatase activity in granulosa cells which always preceded any reduction in the thecal steroidogenic response to LH. It was estimated that between 20 and 60 antral follicles (greater than or equal to 2 mm diam.) per cow may respond to LH by synthesizing androgen whereas only 1-3 follicles (greater than 5 mm diam.) have granulosa cells capable of metabolizing androstenedione or testosterone to oestradiol.

Androstenedione↗