Steroid production, cell proliferation, and apoptosis in cultured bovine antral and mural granulosa cells: development of an in vitro model to study estradiol production.

This study was undertaken to characterize the relationship between changes in steroid production, cell cycle activity (ie, cell proliferation) and apoptosis in antral and mural bovine granulosa cells cultured in vitro. This was done to select conditions promoting optimal estradiol production by bovine granulosa cells cultured in completely defined conditions. In the first experiment, antral granulosa cells were cultured over the entire 4 days of the culture period in the presence of either 0, 2, or 10 ng/ml of FSH (chronic conditions) or were maintained under minimal FSH support (0.5 ng/ml FSH) for the first 3 days of culture and then were challenged over the fourth day of culture with either 0, 2, or 10 ng/ml FSH (challenged conditions). Compared with cells exposed to constant FSH levels (chronic conditions), the FSH-induced production of estradiol was higher (P < 0.006) and that of progesterone was lower (P < 0.02) over the last 24 h of culture, when antral granulosa cells were maintained under minimal FSH support during the first 3 days of culture (challenged conditions). In the second experiment, dynamics of estradiol and progesterone productions, conversion of [14C]androstenedione into subsequent steroid metabolites, DNA content, cell cycle activity, and apoptosis (as assessed by flow cytometry) of antral and mural granulosa cells over the first 3 days of culture under minimal FSH support and in response to a challenge with FSH during the last 24 h of culture were evaluated. Estradiol production as well as the conversion of androstenedione into testosterone and estradiol were greater (P < 0.01) in antral than in mural granulosa cells cultured under challenged conditions. A higher proportion of mural than antral granulosa cells were in the proliferative state at the end of culture (P < 0.03). This may be related to the decreased ability of mural cells to produce estradiol. FSH suppressed (P < 0.05) the spontaneous onset of apoptosis in both cell types. These results suggest that functional differences between these two cell compartments need to be considered in studying bovine granulosa cells in vitro. Because of their large (400 to 600%) FSH-induced estradiol production, antral granulosa cells cultured under challenged conditions provide a model that can be used to examine substances for their ability to alter estradiol production and apoptosis in bovine granulosa cells.

Effect of bovine follicular fluid from healthy and atretic follicles on follicle-stimulating hormone-induced production of estradiol by bovine granulosa cells cultured in vitro.

We studied the effects of active factors present in bovine follicular fluid (bFF) from large healthy or atretic follicles on steroidogenic capability of cultured bovine granulosa cells. Pools of bFF were collected from follicles (> 10 mm; abattoir material) and classified individually as being healthy (bFF-healthy) or atretic (bFF-atretic). Pools of jugular plasma were used as controls and were from heifers bled during the growing (plasma-growing) or the regressing (plasma-regressing) phase of follicular dominance. Granulosa cells were cultured in serum-free conditions and under minimal FSH support (.5 ng/mL) for the first 3 d in order to maintain their physiological estradiol production in response to FSH. Effects of addition of bFF and plasma at final concentrations of 0, 1, or 5% on estradiol and progesterone production and on the percentage of apoptotic cells were determined on d 4 of culture following stimulation of granulosa cells with either 2 or 6 ng/mL FSH. In a parallel experiment, evaluation of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activity was measured following addition of bFF. In contrast to plasma, addition of bFF pools decreased (P < .001) the FSH-induced estradiol production. Such suppression occurred in a dose-related manner (P < .05) and to a greater extent (P < .001) following addition of bFF from atretic than from healthy follicles. The FSH-induced progesterone production was not affected (P > . 1) by addition of bFF but was stimulated (P < .05) by that of plasma. Follicle-stimulating hormone decreased (P < .001) the percentage of apoptotic granulosa cells and this effect was further enhanced (P < .001) by addition of 1 or 5% bFF. The source of bFF did not affect (P > . 1) the percentage of apoptotic cells measured at the end of the culture period. On d 4, treatment with bFF increased (P < .001) granulosa cell androstenedione conversion into testosterone. Results of the present study indicate that factors contained in bFF can suppress granulosa cell estradiol production, and the suppressive effect varies according to the degree of atresia of the follicle from which the fluid has been harvested.

Immunoneutralization of transforming growth factor alpha present in bovine follicular fluid prevents the suppression of the follicle-stimulating hormone-induced production of estradiol by bovine granulosa cells cultured in vitro.

Growth factors such as transforming growth factors alpha (TGF alpha) and beta (TGFbeta) secreted by theca cells and present in bovine follicular fluid (bFF) have been implicated in granulosa cell growth and differentiation. We investigated these phenomena using two complementary approaches to evaluate the physiological contribution of TGF alpha and TGFbeta in the control of the FSH-induced estradiol production in bovine granulosa cells from large follicles. Granulosa cells (3 x 10(5) viable cells/cm2) harvested from eCG-treated prepubertal calves were cultured (serum free) in wells containing 500 microl/cm2 of defined Ham's F-12 medium supplemented with 0.5 ng/ml FSH for the first 3 days (37 degrees C; 95% air:5% CO2). In the first approach, the effects of individual addition of TGF alpha and TGFbeta at final concentrations of 1 x 10(-4) to 10 ng/ml were determined on Day 4 of culture after stimulation of granulosa cell estradiol production with 6 ng/ml FSH. In a second approach, TGF alpha or TGFbeta was removed specifically from bFF (from large follicles > 10 mm) by immunoneutralization. Thereafter, effects of immunoneutralization of TGF alpha or TGFbeta (0, 0.1, 1, 10, and 100 microg/ml anti-TGF neutralizing antibody) present in bFF (2%) were determined on Day 4 of culture following stimulation of granulosa cell estradiol production with 6 ng/ml FSH. On Day 4, FSH increased (p < 0.001) granulosa cell estradiol production (0 vs. 6 ng/ml FSH). Addition of TGF alpha decreased the granulosa cell estradiol production after 6 ng/ml FSH stimulation in a dose-dependent manner (p < 0.001). In contrast, addition of TGFbeta had no effect on the granulosa cell estradiol production (p > 0.1) after the addition of 6 ng/ml FSH. Addition of bFF (2%) decreased (p < 0.0001) the FSH-induced estradiol production by bovine granulosa cells. After immunoneutralization of TGF alpha in bFF, however, this suppressed FSH-induced estradiol production was restored to levels obtained in the absence of bFF, and this occurred in a dose-dependent manner (p < 0.05). Immunoneutralization of TGFbeta in bFF did not prevent (p > 0.1) the suppressive effect of bFF on FSH-induced estradiol production. These results suggest that TGF alpha produced in vivo by large bovine follicles can act locally (auto/paracrine manner) to suppress granulosa cell estradiol production.

Follicle-stimulating hormone-induced estradiol and progesterone production by bovine antral and mural granulosa cells cultured in vitro in a completely defined medium.

Functional subpopulations of granulosa cells exist in bovine follicles. This study was designed to compare the in vitro steroid production by cultured bovine antral and mural granulosa cells in response to various amounts of FSH. Antral and mural granulosa cells (600,000 viable cells/well) harvested from ovaries of PMSG-treated prepuberal calves were cultured in serum-free conditions for 4 d in wells containing 1 mL of defined Ham's F-12 medium, supplemented with 0, 2, or 10 ng/mL of FSH. Culture medium was collected and replaced each day. The mean concentration of estradiol in culture media of bovine granulosa cells decreased from d 1 to d 4 (P < .001). Granulosa cell production of estradiol increased in antral cells following addition of 2 ng/mL FSH (P < .001) but decreased following addition of 10 ng/mL FSH (P < .001) as determined on d 4 by RIA and thin layer chromatography. In contrast, there was no response to FSH stimulation in mural granulosa cells. Progesterone production increased (P < .01) in a dose-dependent manner following stimulation with 2 or 10 ng/mL FSH and was consistently higher (P < .001) in antral than in mural granulosa cells. Addition of LH on d 4 stimulated estradiol and progesterone production in antral (P < .01) but not in mural cells (P > .10). This suggests that FSH- and LH-induced estradiol and progesterone productions are considerably lower in mural than in antral bovine granulosa cells. This suggests that functional differences between these two cell compartments need to be considered in studies involving in vitro cultures of bovine granulosa cells.

Changes in morphological appearance and functional capacity of recruited follicles in cows treated with FSH in the presence or absence of a dominant follicle.

This study was designed to determine the effect of the presence of a dominant follicle at the beginning of FSH stimulation on the morphological appearance and functional capacity of recruited follicles during FSH stimulation in cattle. Synchronized nonlactating dairy cows were assigned to 1 of 2 groups and treated with FSH in the presence (n = 5) or absence (n = 6) of a dominant follicle between Days 7 and 12 of the estrous cycle (Day 0 = estrus) to stimulate follicular growth. Dominant follicles were identified by daily ultrasonographic observations, beginning on Day 3 of the estrous cycle. Dominant follicle had an ultrasonographic diameter > or = 10 mm and were in a growing phase, or maintaining a constant diameter (> or = 10 mm) for less than 4 d. Ovaries were collected at slaughter on the morning of the third day following initiation of the FSH stimulation. All follicles > 2 mm were dissected, classified according to diameter (Class 1: 2 to 4.4 mm; Class 2: 4.5 to 7.9 mm; Class 3: > 8 mm), and incubated individually for 90 min in medium M-199 (37 degrees C, 5% CO2). Following incubation, integrity of each follicle was evaluated histologically to assess the level of atresia and biochemically to determine the in vitro release of estradiol (E2) and androstenedione in culture media. On Day 3 of the FSH treatment, mean number of follicles in each class was similar (P > 0.1) between the 2 groups. The percentage of atretic follicles in Classes 1 and 3 on Day 3 of the FSH stimulation did not differ (P > 0.1) between the 2 groups. However, the percentage of atretic follicles in Class 2 was higher (P < 0.005) in cows treated with FSH in presence than in absence of a dominant follicle (60.8 vs 38.2%). The release of E2 in culture media by small Class 1 atretic or healthy follicles, by Class 2 atretic and by Class 3 healthy follicles was not affected (P > 0.1) by the ovarian status. However (P < 0.001), the release of E2 in culture media of Class 2 healthy and Class 3 atretic follicles was less for follicles harvested from cows bearing than from those not bearing a dominant follicle. Within each follicular class, concentrations of androstenedione in the culture media did not differ between the 2 groups (P > 0.1). These results suggest that the presence of a dominant follicle at the beginning of FSH stimulation alters the population of follicles recruited FSH stimulation. This may be associated with the reported decrease of the superovulatory response in cows superovulated in presence of a dominant follicle.

Ovarian follicular development and endocrine responses in follicular-fluid-treated and hemi-ovariectomized heifers.

The effects of charcoal-extracted bovine follicular fluid (BFF) on endocrine profiles and follicular development in intact and hemiovariectomized postpubertal heifers were examined. Oestrus-synchronized heifers received Norgestomet implants on day 1 and 7 of treatment and were then injected s.c. with 11 ml saline (control) or 11 ml BFF twice a day for 12 days. The ovary bearing the largest follicle (OV1) was removed on day 7 and the remaining ovary (OV2) was collected on day 13. Follicles were observed by daily ultrasonography and were classified according to diameter (size 1: 2-3 mm; size 2: 4-6 mm; size 3: 7-10 mm; size 4: > 10 mm). After ovariectomy they were classified by diameter and histologically as normal or atretic. Intact control heifers had increased numbers of size 4 follicles on OV1 on days 6 and 7; no increase was observed in BFF-treated heifers (P < 0.03). In BFF-treated heifers, the mean basal LH concentration was higher (P < 0.05) and that of FSH was lower (P < 0.04) than in controls. FSH concentrations in BFF-treated heifers decreased from 0.60 +/- 0.08 ng ml-1 (day 1) to 0.22 +/- 0.05 ng ml-1 (day 7; P < 0.04). The concentration of oestradiol increased in control heifers, but not in BFF-treated heifers (P < 0.001). After hemicastration, OV2 underwent compensatory hypertrophy in control heifers, with an increase in the number of size 2, 3 and 4 follicles (P < 0.05), whereas BFF-treated heifers did not. Thus, total follicular volume was much lower in BFF-treated than in control heifers on day 13 (92.2 +/- 15.4 versus 1393.8 +/- 276.6 mm3; P < 0.0002). A transient increase in FSH (P < 0.006) and oestradiol (P < 0.01) concentrations occurred after hemiovariectomy in control but not in BFF-treated animals. In control heifers, an analysis of temporal relationships showed negative correlations between the volume of size 3 and size 4 follicles, and between FSH concentrations and the volume of size 3 and 4 follicles. A positive correlation was found between the mean diameter of the largest follicle and the concentration of oestradiol, whereas negative relationships were found between the concentrations of FSH and oestradiol, and between FSH and the mean diameter of the largest follicle. Analysis of the histological data showed that the number and volume of follicles > 8.57 mm was lower in the BFF-treated OV1 ovary, whereas no differences were found for follicles < or = 8.57 mm.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationships between the level of atresia and inhibin contents (alpha subunit and alpha-beta dimer) in morphologically dominant follicles during their growing and regressing phases of development in cattle.

The objectives were to determine the relationships between changes in the levels of histological and biochemical (estradiol [E2]:androstenedione [A], E2:A ratio) atresia and changes in inhibin contents of morphologically dominant follicles collected during the growing or the regressing phase of the first wave of follicular development in cycling crossbred beef heifers. Heifers were slaughtered either when the dominant follicle (> or = 9 mm; diameter of the antral cavity as assessed by ultrasonography) of the first wave was still growing (n = 7) or when the first dominant follicle (> or = 9 mm; n = 7) was regressing or was at the end of the plateau phase. Following ovary collection, the dominant follicle was dissected and level of histological atresia was determined on sections of follicular walls. Aliquots of follicular fluid from each of the dominant follicles were collected to measure concentrations of E2, A, and inhibin alpha subunit by RIA and to measure concentrations of dimeric (alpha-beta dimer) inhibin by a two-site immunoradiometric assay. Heifers were slaughtered on Days 5.4 +/- 0.5 (growing phase) and 11.8 +/- 0.5 (regressing phase) of the estrous cycle, and mean diameter of the dominant follicles was similar in both phases (9.9 +/- 0.4 vs. 10.8 +/- 0.4 mm; p > 0.09). All morphologically dominant follicles collected during the growing phase (7/7) were histologically healthy and estrogen-active (E2:A ratio > 1), while those collected during the regressing phase (7/7) were histologically atretic and estrogen-inactive (E2:A ratio < 1; chi 2 = 14.0, p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)

Histochemical and immunocytochemical characterizations of laminated bodies in the pancreas acinar lumen.

Combined adrenalectomy and castration cause numerous morphological alterations in the exocrine pancreas of the rat. Accumulation of laminated bodies (LB) in the acinar lumen is one of these alterations. A series of classical histological stains was applied to identify the components of these structures. A positive reaction was observed with periodic acid-Schiff, Congo red, and Alcian blue (pH 2.5). The first stain reacts with neutral and some acid mucosubstances and the second with amyloid, whereas the last one reacts with sulfated and nonsulfated acid mucosubstances. The LB also responded to Luxol fast blue, indicating the presence of lipids, an observation that is in agreement with the osmiophilic properties of these structures. A more specific identification of LB components was carried out with the immunocytochemical protein A-gold technique. Presence of gamma-glutamyltranspeptidase (gamma-GT) and GP2, two glycoproteins known to be secreted by the pancreas, was tested. The gamma-GT was associated with LB whereas GP2 was found in the lumen but not associated with these structures. Amylase was undetectable when LB occupied the lumen, suggesting that the process leading to production of LB also blocks secretory activity. To determine if diet influences LB accumulation in the pancreas acinar lumen, their frequency was compared in rats fed Purina Lab Chow or a lipid-free synthetic diet. A significant increase was observed in castrated-adrenalectomized rats fed the latter diet. This increase corresponded to a pronounced reduction in the number of zymogen granules (ZG) in the acinar cell. Our results show that LB are made of lipids, neutral mucosubstances, and nonsulfated acid mucosubstances, and that hormonal (steroids) and dietary factors (lipids) influence their accumulation.

Effects of pregnant mare's serum gonadotropin treatment on follicular populations and ovulation rates in prepuberal gilts with two morphologically different ovarian types.

Two experiments were conducted to determine the effects of pregnant mare's serum gonadotropin (PMSG) on follicular populations and ovulation rates in prepuberal gilts with grape-type (GT) and honeycomb-type (HT) ovaries. The follicular populations were determined at 170 d of age (d 0) and 19 d after PMSG (d 19). In Exp. 1, the mean number of macroscopic follicles of Classes 2 and 3 was greater (P less than .05) in GT (n = 11) than in HT (n = 32) ovaries at d 0, whereas the mean number of those of Class 1 was greater (P less than .05) in HT ovaries. At d 19, no difference was observed between the two ovarian types for any class of follicles. The PMSG-induced ovulation rates were comparable between the two ovarian types (8.3 vs 7.9, GT vs HT, respectively; P greater than .10). In Exp. 2, the microscopic follicular populations were determined on right and left ovaries removed, respectively, on d 0 and 19. At d 0, GT ovaries (n = 5) contained a greater number of Class 5 nonatretic (P less than .01) and atretic (P less than .02) follicles than did HT ovaries (n = 5), whereas at d 19 the mean number was not statistically different between the two ovarian types (P greater than .10). In contrast, gilts with HT ovaries contained a greater (P less than .01) number of Class 4 atretic follicles than gilts with GT ovaries at d 0, whereas at d 19 the mean number was not statistically different between the two ovarian types (P greater than .10).(ABSTRACT TRUNCATED AT 250 WORDS)

Pituitary LH content and plasma LH levels following daily GnRH analogue treatment in male red-winged blackbirds (Agelaius phoeniceus).

1. During the 14-day treatment period, plasma LH levels following GnRH analogue (GnRH-A) injections (10.0 micrograms) were significantly reduced after the 6th and the 14th injection. 2. One day post-treatment, the LH pituitary content was significantly reduced in GnRH-A-treated redwings compared to saline-injected controls. 3. Pituitary LH content was significantly higher in GnRH-A treated birds compared to control birds 14 and 28 days post-treatment and plasma LH levels were similar in both groups. 4. Hypersecretion of LH following GnRH-A injections appears to reduce pituitary LH content, acting as a stimulus for its synthesis. 5. These results suggest a higher LH synthesis and storage in the pituitary gland of the GnRH-A-treated birds compared to the control birds during the post-treatment period.

Vascularized bone transfer for tibial pseudarthrosis. Part I: Vascularized fibula transplantation. A report of 4 cases in the treatment of traumatic bone defects of the tibia.

The limited experience of reconstruction of four cases of large tibial defects with free vascularized fibular grafts is presented. Healing occurred in three of the four patients, and one developed a distal pseudarthrosis. One graft fractured twice but healed with a hypertrophic callus.

Ovarian morphological conditions and the effect of injection of human chorionic gonadotropin on ovulation rates in prepuberal gilts with two morphologically different ovarian types.

The purpose of this experiment was to determine the ovulation rate after treatment with human chorionic gonadotropin (hCG) in two groups of gilts characterized by different ovarian morphology: grape-type (GT; n = 11) and honeycomb-type (HT; n = 7). At 170 d of age (d 0), gilts were examined by laparoscopy and ovarian type was determined by the distribution of macroscopic follicles present on the ovarian surface. Five to ten minutes after surgery, each gilt received a single injection (i.m.) of 750 IU of hCG. At d 0, GT ovaries had a greater number of large follicles (greater than or equal to 6 mm) than HT ovaries (10.0 +/- .5 vs 2.6 +/- .3; P less than .05), whereas HT ovaries had more small follicles (1 to 3 mm; HT: 42.3 +/- .8 vs GT: 26.7 +/- .9; P less than .05) and total follicles (HT: 59.4 +/- 2.3 vs GT: 52.2 +/- 1.5; P less than .05), although numbers of medium follicles (4 to 5 mm) were similar (GT: 15.6 +/- .8 vs HT: 14.6 +/- 1.7; P greater than .10). Number of induced corpora lutea (CL) per ovary was greater (P less than .05) in gilts with GT ovaries (10.59 +/- 2.9 CL) than in gilts with HT ovaries (5.21 +/- .66 CL). Total weight of luteal tissue (LT) per ovary and serum progesterone concentrations 8 d after induction of ovulation were greater in GT gilts than in HT gilts (GT: 6.37 +/- 1.09 g vs HT: 3.31 +/- .49 g for LT, P less than .05; GT: 21.08 +/- 4.76 ng/ml vs HT: 13.40 +/- 2.05 ng/ml for progesterone, P less than .07).(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma LH and androgen levels in the red-winged blackbird (Agelaius phoeniceus) treated with a potent GnRH analogue.

1. Low doses of GnRH-A (0.01-0.10-1.0 micrograms) given during the annual testes growth period did not clearly affect plasma LH and androgen levels 10 min following the injection. 2. The first injection of high doses of GnRH-A (2.0-10.0-20.0 micrograms) markedly increased plasma LH and androgen levels measured 10 min following the injection. The increase in plasma LH level was dose-dependent and the maximal LH level was obtained with 10.0 micrograms of GnRH-A. 3. Impairment of the LH response to GnRH-A was assessed by comparing the first and the fourteenth injection of high doses of GnRH-A. Evidences of pituitary gland desensitization are reported since plasma LH levels were reduced following the fourteenth injection in all groups. 4. Plasma androgen levels following high doses of GnRH-A were not clearly affected in red-winged blackbirds.

Decreased superovulatory responses in heifers superovulated in the presence of a dominant follicle.

Dairy heifers were superovulated in the presence (dominant group, N = 8) or absence (non-dominant group, N = 6) of a dominant follicle at the start of a a superovulatory treatment on Days 7-12 of the oestrous cycle (Day 0 = oestrus). Daily ultrasonographic observations of ovaries (recorded on videotape) starting on Day 3 were used to assess the presence or absence of a dominant follicle (diameter greater than 9 mm, in a growing phase or at a stable diameter for less than 4 days) and to monitor follicular development before and during treatment. The number of CL estimated by ultrasonography (7.1 +/- 1.8 vs 13.5 +/- 1.4) or by rectal palpation (6.9 +/- 2.0 vs 16.3 +/- 1.6) and mean progesterone concentrations (32.5 +/- 19 vs 80.7 +/- 16 ng/ml) after treatment were lower (P less than 0.01) in the dominant than in the non-dominant group. Based on number of CL, two populations of heifers were identified in the dominant group, i.e. those that had a high (dominant-high, N = 4; greater than 7 CL) or a low (dominant-low, N = 4; less than 7 CL) response to treatment. During treatment, the increases in number of follicles 7-10 mm and greater than 10 mm in diameter occurred sooner and were of higher magnitude in the non-dominant than in the dominant-high or dominant-low groups (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Bovine borreliosis: comparison of simple methods for detection of the spirochaete in the blood.

Natural Borrelia infection occurring in a calf was followed up using stained blood and buffy coat smears and direct observation of blood and buffy coat in order to compare these spirochaete detection methods. Direct examination of the platelet-plasma interface of the microhaematocrit capillary tubes after centrifugation allowed detection of two parasitaemias that were not seen in Giemsa stained blood smears. An ascending Babesia bigemina infection seemed to suppress the Borrelia parasitaemia. Spirochaetes were detected in five out of 37 Boophilus decoloratus ticks recovered from the infected calf. The blood and the haemolymph stages of Borrelia were measured. After treatment with oxytetracycline the spirochaete could no longer be detected.

Influence of pregnancy on the onset of oestrus and luteal function after prostaglandin-induced luteolysis in cattle.

Luteolysis was induced by an injection of 500 micrograms cloprostenol (a prostaglandin (PG) analogue) in pregnant (P) Holstein heifers on Days 17 or 24 of gestation and in non-pregnant (NP) Holstein heifers on Day 17 of the oestrous cycle (oestrus = Day 0). Heifers in Groups P-17 (N = 8) and P-24 (N = 8) were inseminated twice whereas those in Group NP-17 (N = 8) were not inseminated. Immediately after PG injection, embryos were recovered by uterine flushing (400 ml) to confirm pregnancy in Groups P-17 and P-24. Uterine flushing with an equivalent volume of physiological saline was also done in Group NP-17. The interval from PG injection to oestrus and to the peak of luteinizing hormone (LH) as well as profile of increase in plasma oestradiol concentrations during that period did not differ (P greater than 0.1) among the groups. However, the proportion of heifers exhibiting abnormal luteal phases (primarily of short duration) during the oestrous cycle after PG injection was greater (P less than 0.01) in Group P-24 than in Groups NP-17 + P-17 pooled (6/8 vs 3/16). These results suggest that the previous presence of a conceptus did not have any effect on the onset of oestrus, or on plasma concentrations of oestradiol and LH after PG-induced luteolysis on Days 17 or 24 of gestation. However, luteal function during the subsequent oestrous cycle was impaired if heifers were 24 days pregnant when luteolysis was induced.

Effects of litter size and light regimen imposed during lactation on ovarian follicular population at weaning in third-parity sows.

Sixteen third-parity sows maintained under two photoperiod treatments (16 h light: 8 h dark [16 h] or 4 h light: 20 h dark [4 h]) were assigned to be unilaterally ovariectomized (left ovary) at weaning on d 28 after farrowing a small litter (12 or fewer piglets; S) or a large litter (more than 12 piglets; L). Antral follicles were classified as atretic or nonatretic and divided into six size classes. The right ovary was obtained at slaughter, 16 d after estrus was detected. Number of antral follicles in the left ovary was greater (P less than .05) for L sows (540) than for S sows (427). Sows exposed to 16 h of light with large litters had a higher number of atretic follicles than all other groups (litter size x photoperiod interaction, P less than .05). Number of nonatretic follicles were similar between groups. However, when nonatretic follicles were classed by size (1 = smallest; 6 = largest), their distribution differed. In Class 1, L sows had more follicles than S sows (P less than .05). For Classes 3 and 4, photoperiod interacted with litter size to decrease the number of follicles in L 16 h sows (litter size x photoperiod interaction, P less than .05). These lower numbers of nonatretic follicles were related to a higher rate of follicular atresia in L 16 h sows. In Classes 2, 3 and 4, L 16 h sows had more atretic follicles than any other group (litter size x photoperiod interaction, P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)