Vascularity and expression of angiogenic factors in bovine dominant follicles of the first follicular wave

To determine the relationships among vascularity, expression of angiogenic factors, and selected intrafollicular factors in dominant and nondominant follicles of the first follicular wave, ovaries were obtained on d 3 of the estrous cycle from mature cross-bred beef heifers (n = 8) after a synchronized estrus. Follicular fluid (FF) was collected from all follicles > or = 3 mm for determination of estradiol-17beta (E), progesterone (P4), vascular endothelial growth factor (VEGF), and IGFBP concentrations. The ovaries were then perfusion-fixed and used for histochemical detection of lectin BS-1 (a marker of endothelial cells and thus vascularization) binding, and immunolocalization of VEGF, endothelial nitric oxide synthase (eNOS), and proliferating cell nuclear antigen, followed by image analysis of selected follicles. Follicles were classified, based on E and P4 concentrations in FF, as dominant, estrogen-active (EA; E:P4 > or = 1) or nondominant, estrogen-inactive (EI; E:P4 <1). Concentrations of E and VEGF in FF, the area of positive staining for lectin BS-1, VEGF, and eNOS, and the labeling index (an index of the percentage of cells proliferating) in granulosa and theca layers were greater (P < 0.05) in the EA than in the EI follicles, but concentrations of P4 and IGFBP in FF were less (P < 0.05) in EA than in EI follicles. In addition, vascularity was positively correlated (P < 0.05) with VEGF and eNOS protein expression, and tended (P < 0.1) to be positively correlated with the E:P4 ratio in FF but tended (P < 0.1) to be negatively correlated with IGFBP and P4 concentrations in FF. These data highlight the importance of vascularity, angiogenic factors, and IGFBP in the health of the dominant follicle in heifers, and indicate that the FF concentrations of E, VEGF, IGFBP, and P4, and the E:P4 ratio can be used as markers of dominant follicles.     

Relationship between two morphological types of ovaries and their follicular microscopic population before puberty and their ensuing follicular development and ovulation rates at puberty in gilts

A population of ovarian follicles was studied in 32 gilts with two morphological types at four physiological stages. Left ovaries were serially sectioned (10 microns), and follicles (greater than .19 mm) were counted and measured by histological technique. In Exp. 1, prepuberal gilts that maintained the grape-type (GT) or other-type (OT) of ovaries at 140 and 160 d of age as determined by laparoscopy had their population of follicles studied at 165 d of age and compared with that on d 3 of the first puberal estrus in gilts that maintained the same ovarian type (GT or OT) at 140, 160 and 180 d of age. In Exp. 2, gilts that maintained the same ovarian types at 140, 160 and 180 d of age were compared at d 19 and 3 of the first and second puberal estrus, respectively. At 165 d of age, OT ovaries contained a greater number of classes 3 (.63 to 1.12 mm) and 4 (1.13 to 2.00 mm) nonatretic (less than 4 pycnotic bodies) follicles than GT (P less than .01), whereas at d 3 of the first puberal estrus, their mean number became comparable between the two ovarian types (P greater than .1) and similar to 165-d GT ovaries. At 165 d of age, atretic follicles formed 19.6% and 10.7% of the population of antral follicles in OT and GT ovaries (P less than .05) whereas at puberty 17.5% and 19.6% atresia was observed in the same two groups, respectively (P greater than .10).(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of in vitro development of embryos collected from the same gilts at first and third estrus

In vitro development of embryos collected from the same gilts mated at first and third estrus was compared. Embryos from one to eight cells were collected from gilts 36 to 48 h after detection of estrus. Embryos were cultured for 8 d in Whitten's medium in a humidified atmosphere of 5% CO2 in air at 37 degrees C and were observed daily. No differences were detected among percentages of one- to eight-cell embryos developing into morulae from gilts in first or third estrus (P greater than .05). Similar percentages of one- to two-cell embryos from gilts mated at first and third estrus developed into blastocysts (45.8 and 55.2%, respectively), expanded blastocysts (10.4 and 24.1%, respectively) and hatching blastocysts (4.2 and 3.4%, respectively; P greater than .05). Fewer three- to eight-cell embryos from gilts in first estrus than from gilts in third estrus developed into blastocysts (63.4 and 91.1%) and expanded blastocysts (14.6 and 55.6%; P less than .01). Similar percentages of embryos with abnormal morphology were observed among morulae developing from one- to eight-cell embryos collected from gilts mated at first and third estrus (14.9 and 9.9%, respectively; P greater than .05). In contrast, more morphologically abnormal embryos were observed among blastocysts developing from gilts mated at first estrus than at third estrus (31.2% and 14.0%, respectively; P less than .05). The results suggest that the reduced in vitro development of embryos collected from gilts mated at first estrus may be due to an aberration in blastocoel formation and expansion.     

The levels of 5alpha-dihydrotestosterone in follicular fluid in healthy and atretic ovine follicles

Dihydrotestosterone (DHT) induces follicular atresia under experimental conditions. However, whether it causes any antagonistic effect under natural condition is not known. In the present study, we investigated concentrations of DHT in follicular fluid and correlated them with concentrations of estradiol-17beta (E2) and its androgen substrates, androstenedione (A4) and testosterone (T), in healthy and atretic follicles of sheep. Merino ewes were treated twice with PGF2alpha (PG) to synchronize estrus. The ovaries were recovered at 14 days after the second PG (luteal phase) or 24h after the third PG given 14 days after the second PG (follicular phase). Follicles were dissected and their size and appearance were recorded. Follicular fluid was collected from follicles larger than 3.5mm and concentrations of E2, progesterone (P4), A4, T and DHT were determined by RIA. The inhibitory effect of DHT on conversion of T to E2 was tested in cultured granulosa cells. Appreciable levels of DHT were observed in the follicular fluid of ovine preovulatory follicles. The levels of DHT were much lower than those of E2, A4 and T, irrespective of physiological conditions of follicles. No difference was found in DHT concentration between healthy and atretic follicles. Dihydrotestosterone marginally inhibited aromatization of T in granulosa cells but this effect was only observed when the levels of DHT were 10 times higher than that of T in culture medium. These results indicate that DHT is present in ovine preovulatory follicles but its levels are not sufficient to exert any antagonistic effect on follicular development.     

Ovarian response to pregnant mare serum gonadotropin and porcine pituitary extract in gilts actively immunized against gonadotropin releasing hormone

Two experiments were conducted to determine the effect of exogenous gonadotropins on follicular development in gilts actively immunized against gonadotropin releasing hormone (GnRH). Four gilts, which had become acyclic after immunization against GnRH, and four control gilts were given 1,000 IU pregnant mare serum gonadotropin (PMSG), while four additional control gilts were given saline. Control animals were prepuberal crossbred gilts averaging 100 kg body weight. Control gilts given saline had ovaries containing antral follicles (4 to 6 mm in diameter). Control gilts given PMSG exhibited estrus and their ovaries contained corpora hemorrhagica and corpora lutea. PMSG failed to stimulate follicular growth in gilts immunized against GnRH, and ovaries contained regressed corpora albicantia and small antral follicles (less than 1 mm in diameter). Concentrations of luteinizing hormone (LH) and estradiol-17 beta (E2) were non-detectable in gilts immunized against GnRH and given PMSG. In the second experiment, five gilts actively immunized against GnRH were given increasing doses of PMSG every third day until unilateral ovariectomy on d 50. PMSG failed to stimulate follicular growth, and concentrations of follicle stimulating hormone (FSH), E2 and LH were not detectable. Six weeks later, gilts were given a booster immunization and then were given 112 micrograms LH and 15 micrograms FSH intravenously every 6 h for 9 d. The remaining ovary was removed on d 10. Although LH and FSH concentrations were elevated, administration of gonadotropins did not stimulate follicular growth or increase E2 concentrations. These results indicate that neither PMSG or exogenous LH and FSH can induce E2 synthesis or sustain follicular development in gilts actively immunized against GnRH.     

Study of follicular development and heredity of morphological types of ovaries in prepuberal gilts

Two different morphological types of ovaries ("grape" and "other" types) previously described by microscopic examination were studied on 107 live gilts. The ovarian type and distribution of macroscopic follicles during the prepuberal period were determined at 140, 160 and 180 d of age by laparoscopic examination. The incidence of grape-type (GT) ovaries was higher (P less than .05) at 140 d of age (50.5%) than at 160 d (38.3%) or 180 d (35.5%). At maturity (d 19 of the first postweaning estrus), 37.7% of the ovaries were classified as GT. The proportion of gilts that maintained their initial type (at 140 d of age) at 160 d, 180 d and at maturity was higher (P less than .05) for the gilts having the other-type (OT) ovaries (49% and 25% for OT and GT, respectively). The mean number of small follicles (1 to 3 mm in diameter) was higher (P less than .05) for the OT ovaries at 140 d (71.5 vs 18.0), at 160 d (79.3 vs 26.2) and at 180 d (77.0 vs 32.8). The number of follicles of 4 to 5 mm was higher (P less than .05) for GT ovaries at 140 d (16.4 vs 4.8), at 160 d (20.9 vs 4.8) and at 180 d of age (21.4 vs 5.7). The large follicles (greater than or equal to 6 mm) were more numerous for the GT at 140 d (7.0 vs .3), at 160 d (5.7 vs .5) and at 180 d (4.4 vs .6). No statistical difference was observed at maturity in the distribution of follicles between the two types.(ABSTRACT TRUNCATED AT 250 WORDS)     

Follicular fluid concentrations of estradiol-17 beta and progesterone and secretory patterns of LH and FSH in prepubertal gilts reared in confinement or outdoor lots

One-hundred-twenty crossbred gilts from two experiments were assigned randomly to a 2 X 5 factorial experiment. Gilts were reared in two environments (confinement or outside) and assigned to be slaughtered at 4, 5, 6, 7 or 8 mo of age. Beginning at 6 mo of age, blood samples were taken at weekly intervals from each gilt via venipuncture. Serum concentrations of progesterone were analyzed to determine when gilts attained puberty. On the day prior to slaughter, six pigs within a treatment group were cannulated and blood samples were taken at 20-min intervals for 4 h. At slaughter, follicular fluid (FF) was aspirated and the volume determined from those follicles having a diameter of at least 4 mm. No effect of environment was found on the proportion of gilts that attained puberty by 8 mo of age. For the 12 gilts that reached puberty during the study, the age at puberty for gilts reared in outdoor lots (202 +/- 5 d) was less (P less than .05) than those reared in confinement (224 +/- 8 d). Mean concentrations of serum luteinizing hormone (LH; P = 98) and number of secretory spikes of LH (P = .76) were similar between gilts reared in confinement and those reared in outdoor lots. No differences in average serum concentrations of follicle stimulating hormone (FSH) or number of secretory spikes of FSH were found between gilts subjected to these environments (P = .95). Concentrations of estradiol-17 beta in FF were not affected by environment or age (P greater than .25).(ABSTRACT TRUNCATED AT 250 WORDS)     

Factors contributing to early embryonic mortality in gilts bred at first estrus

Gilts bred at first (n = 18) and third (n = 18) estrus were assigned in replicates of equal numbers to be slaughtered on d 3, 15 and 30 post-mating to assess fertilization rate, embryonic losses and serum concentrations of estrogen (estradiol-17 beta + estrone) and progesterone. Mean number of ovulations was lower among gilts bred at first vs third estrus (12.2 vs 14.5; P less than .05), with no difference in fertilization rate (100 vs 98%). Embryonic survival was lower (P less than .05) among gilts bred at first vs third estrus on d 15 (78.1 vs 95.4%) and 30 (66.7 vs 89.4%) of gestation. Serum estrogen (pg/ml) and progesterone (ng/ml) levels, although lower in gilts bred at first vs third estrus, were not significantly different at the three stages of gestation studied. The ratio of progesterone to estrogen in gilts bred at first estrus was higher than in those bred at third estrus on d 15 (439 +/- 71 vs 210 +/- 17) and 30 (597 +/- 106 vs 179 +/- 50), but was lower on d 3 (187 +/- 37 vs 444 +/- 123; stage of gestation X estrous period interaction, P less than .05). These data suggest that changes in the ratio of systemic levels of estrogen and progesterone may be related to early embryonic mortality in gilts bred at pubertal estrus.     

Ovarian function and hormone secretion of gilts actively immunized against androstenedione

Fifty crossbred gilts immunized against bovine serum albumin (BSA) or androstenedione conjugated to BSA (AD) were used in three experiments. Primary immunizations were given at 120 d of age and boosters at 148 and 176 d. Gilts were moved to pens containing four to five animals each and exposed to boars beginning at 180 d of age. Immunization against AD did not affect age at puberty, percentage of gilts exhibiting estrus or duration of first estrous cycle. Over the three experiments, ovulation rate was 24% greater for AD-immunized gilts than for controls, and the number of corpora lutea was related positively (r = .82) to the log of the antibody titer. Number of ovulations decreased as interval from booster immunization to onset of estrus increased. During diestrus of the first estrous cycle, gilts immunized against AD had more follicles 5 to 10 mm in diameter, more total ovarian follicles and more total ovarian structures (corpora lutea plus follicles) than controls. Immunization against AD increased the frequency of LH pulses on d 16 but not on d 17 or 18, of the estrous cycle. However, average serum concentrations of LH, FSH and estradiol from 5 d before until 2 d after expected estrus were not different between treatment groups. Concentrations of AD in follicles 4 to 6 and greater than 7 mm in diameter were greater in gilts immunized against AD. Mean serum progesterone was higher on d 9 and 12 after mating in AD immunized gilts than in controls. Immunization against AD had no effect on maintenance of pregnancy or embryo survival rate.     

Estrogen receptors in the uterus and ovarian follicles of gilts treated with dihydrotestosterone

Experiments were conducted to evaluate expression of the estrogen receptor (ER) alpha and ERbeta genes in the uterus and ovarian follicles of gilts treated with 5alpha-dihydrotestosterone (DHT) during the follicular phase of the estrous cycle. This DHT treatment has enhanced ovulation rate but decreased blastocyst survival in previous experiments. Gilts received daily i.m. injections of 10 mg of DHT from day 13 (day 0 = onset of estrus) to day 18 (experiment 1), or from day 13 to 16 (experiment 2) of the estrous cycle. Gilts that served as controls received vehicle. The ovaries and a portion of uterine horn were surgically removed 24 h after the last treatment. Administration of DHT from day 13 to 18 of the estrous cycle decreased uterine wet weight (tendency, P = 0.10), and the relative amounts (ratios to ribosomal protein L19) of endometrial mRNA for the estrogen-responsive gene complement component C3. Gilts receiving DHT had greater amounts of ERbeta mRNA in the endometrium than those treated with vehicle in both experiments, but DHT did not alter the overall amounts of endometrial ERalpha mRNA. Immunohistochemical (IHC) analysis demonstrated that DHT did not alter the relative amounts of ERalpha in the myometrium, glandular and luminal epithelia and endometrial subepithelial stroma. In the ovary, amounts of ERalpha and ERbeta mRNAs in surface walls of follicles > or =6 mm in diameter were not altered by DHT treatments, however, DHT treatment from day 13 to 16 decreased the amounts of immunoreactive ERalpha in the theca interna at the surface walls of day 17 follicles (experiment 2). The amounts of immunoreactive ERalpha were greater in the granulosa than in the theca interna, and within cell type, the amounts of ERalpha were greater at the surface than at the basal region of the follicles, with the exception of the theca interna in follicles evaluated on day 19 (experiment 1). Treatment of gilts with DHT during the follicular phase of the estrous cycle increased ERbeta mRNA in the endometrium and influenced the amounts of immunoreactive ERalpha in ovarian follicles in a cell type-, day of development- and region-specific manner.     

Follicular development and maturation in gilts selected for an index of high ovulation rate and high prenatal survival

Seventy-one 10th-generation gilts from White Line-1 (WL-1 = randomly selected control line) and White Line-2 (WL-2 = selected for an index of ovulation rate and prenatal survival rate) were used to compare the pattern of follicular development and atresia during the follicular phase of the estrous cycle. Gilts were treated with PGF(2alpha)on d 13 of the estrous cycle (d 0 of induced follicular development) to induce luteolysis and assigned randomly within line and sire for ovary recovery on d 0, 2, 3, 4, 5, and the day after estrus. Ovaries were evaluated for numbers of corpora albicantia and small (2 to 2.9 mm), medium (M1 = 3 to 4.9 mm; M2 = 5 to 6.9 mm), and large (>or=7 mm) follicles. The concentration of estradiol-17beta in follicular fluid was used to classify individual M2 and large follicles as estrogen-active (>or=100 ng of estradiol-17beta/mL) or inactive (<100 ng of estradiol-17beta/mL). The WL-2 gilts had a greater ovulation rate than WL-1 gilts at their pre-treatment estrus (20.4 vs. 13.8 corpora albicantia; P < 0.001). The small and M1 follicle populations decreased rapidly in both lines over time (P < 0.001). The M2 follicle population increased in both lines between d 0 to 4 and then decreased. Mean estradiol concentration of M2 follicles increased in both genetic lines over time (P < 0.02). All large follicles were estrogen-active in both lines; the number of large follicles increased with day (P < 0.001) and was similar in both lines. The number of estrogen-active M2 follicles was similar in both lines, increasing to d 3 and 4 and then decreasing (P < 0.01) thereafter. However, the total number of estrogen-active follicles (sum of estrogen-active M2 and large follicles) was greater in WL-2 than in WL-1 gilts (P < 0.04), increasing to the ovulatory potential by d 3 in WL-1 gilts, but continuing to increase through d 4 in WL-2 gilts. Selection of an additional six ovulatory follicles from the estrogen-active M2 follicle pool after d 5 was required in both lines to achieve the projected ovulation rate, and after estrus, the number of large follicles remained insufficient to attain the ovulatory potential of each line.     

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.     

Endocrine changes associated with a dietary-induced increase in ovulation rate (flushing) in gilts

Effects of an increased level of dietary energy (flushing) on plasma concentrations of FSH, LH, insulin, progesterone and estradiol-17 beta and ovulation rate were studied in 16 gilts. Gilts received 5,400 kcal ME/d for one estrous cycle and the first 7 d of a second. On d 8 of the second estrous cycle, gilts received either 5,400 kcal ME/d (control [C], n = 8) or 11,000 kcal ME/d (flushed [F], n = 8) for the remainder of the estrous cycle. Blood was collected daily at 15-min intervals for 6 h from d 8 through estrus. Gilts were examined by laparotomy 6 d after estrus. Ovulation rate was greater (P less than .05) in F than C gilts (16.0 vs 9.4). Mean daily concentrations of FSH were greater (P less than .05) in F gilts at 5 d, 4 d and 3 d prior to estrus compared with C females. In both C and F gilts, FSH decreased (P less than .05) prior to estrus. Mean daily concentrations of LH and LH pulse amplitude were not different (P greater than .05) between treatments. Mean number of LH pulses/6 h at 4 d, 3 d and 2 d prior to estrus were greater (P less than .05) in F than in C gilts. In both treatments, LH pulse amplitude decreased (P less than .05) and pulse frequency increased (P less than .07) prior to estrus. Mean plasma concentrations of insulin tended to be higher (P less than .07) in F than in C females during the 7-d period before estrus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of porcine somatotropin on number of granulosa cell luteinizing hormone/human chorionic gonadotropin receptors, oocyte viability, and concentrations of steroids and insulin-like growth factors I and II in follicular fluid of lean and obese gilts.

Prepubertal gilts of obese (n = 24) or lean (n = 24) genetic lines were injected (s.c.) daily with 0, 2, or 4 mg of porcine somatotropin (pST) for 6 wk starting at 160 d of age to determine whether pST affects follicular function. Blood and ovaries were collected at slaughter 24 h after the last injection. Surface follicles greater than or equal to 1.0 mm in diameter were counted, and pools of follicular fluid (FFL) and granulosa cells were collected from 1.0- to 3.9-mm (small) and 4.0- to 6.9-mm (medium) follicles. Oocytes were collected from small and medium follicles and evaluated for maturational stage and viability. Porcine somatotropin increased (P less than .08) the numbers of small but not the numbers of medium follicles per gilt (P greater than .10). Oocyte maturation and viability were not affected by pST or genetic line. Porcine somatotropin increased (P less than .05) concentrations of insulin-like growth factor I (IGF-I) in serum and FFL of both obese and lean gilts; IGF-I was lower (P less than .01) in lean gilts. Treatment with pST decreased (P less than .05) IGF-II in FFL of lean but not in that of obese gilts. Dose of pST and line had no effect on concentrations of progesterone in FFL of small or medium follicles or on concentrations of estradiol in FFL of small follicles.(ABSTRACT TRUNCATED AT 250 WORDS)     

Flushing and altrenogest affect litter traits in gilts

Gilts (n = 267) were allotted to flushing (1.55 kg/d additional grain sorghum), altrenogest (15 mg.gilt-1.d-1) and control treatments in a 2 x 2 factorial arrangement. Altrenogest was fed for 14 d. Flushing began on d 9 of the altrenogest treatment and continued until first observed estrus; 209 gilts (78%) were detected in estrus. The interval from the last day of altrenogest feeding to estrus was shorter (P less than .05) with the altrenogest + flushing treatment (6.6 +/- .2 d) than with flushing alone (7.6 + .3 d). Ovulation rates (no. of corpora lutea) were higher (P less than .05) in all flushed gilts (14.5 +/- .4 vs 13.4 +/- .4), whether or not they received altrenogest. Flushing also increased the total number of pigs farrowed (.9 pigs/litter; P = .06) and total litter weight (1.43 kg/litter; P = .01), independent of altrenogest treatment. Number of pigs born alive and weight of live pigs were higher for gilts treated with altrenogest + flushing and inseminated at their pubertal estrus than for gilts in all other treatment combinations. In contrast, gilts receiving only altrenogest had greater live litter weight and more live pigs born when inseminated at a postpubertal estrus than when inseminated at pubertal estrus. We conclude that flushing increased litter size and litter weight, particularly for gilts that were inseminated at their pubertal estrus. Increased litter size resulted from increased ovulation rates, which, in nonflushed gilts, limited litter size at first farrowing.     

Effects of active immunization against growth-hormone releasing factor on puberty and reproductive development in gilts.

Hormones within the somatotropin cascade influence several physiological traits, including growth and reproduction. Active immunization against growth hormone-releasing factor (GRFi) initiated at 3 or 6 mo of age decreased weight gain, increased deposition of fat, and delayed puberty in heifers. Two experiments were conducted to investigate the effects of GRFi on puberty and subsequent ovulation rate in gilts. Crossbred gilts were actively immunized against GRF-(1-29)-(Gly)2-Cys-NH2 conjugated to human serum albumin (GRFi) or against human serum albumin alone (HSAi). In Exp. 1, gilts were immunized against GRF (n = 12) or HSA (n = 12) at 92 +/- 1 d of age. At 191 d of age, antibody titers against GRF were greater (P < .05) in GRFi (55.5 +/- 1.3%) than in HSAi (.4 +/- 2%) gilts. The GRFi decreased (P < .05) BW (86 +/- 3 vs 104 +/- 3 kg) by 181 d of age and increased (P < .05) backfat depth (15.7 +/- .4 vs 14.8 +/- .4 mm) by 130 d of age. At 181 d of age, GRFi reduced the frequency of ST release (1.0 +/- .5 vs 5.0 +/- .5, peaks/24 h; P < .0001) and decreased (P < .01) ST (1.1 +/- .06 vs 1.7 +/- .06 ng/mL), IGF-I (29 +/- 2 vs 107 +/- 2 ng/mL), and insulin concentrations (3.5 +/- .2 vs 6.3 +/- .2 ng/mL). The GRFi decreased (P < .05) feed conversion efficiency but did not alter age at puberty (GRFi = 199 +/- 5 d vs HSAi = 202 +/- 5 d) or ovulation rate after second estrus (GRFi = 10.7 +/- .4 vs HSAi = 11.8 +/- .5). In Exp. 2, gilts were immunized against GRF (n = 35) or HSA (n = 35) at 35 +/- 1 d of age. The GRFi at 35 d of age did not alter the number of surface follicles or uterine weight between 93 and 102 d of age, but GRFi decreased (P < .05) ovarian weight (.41 +/- .08 vs 1.58 +/- .4 g) and uterine length (17.2 +/- 1.1 vs 25.3 +/- 2.3 cm). Immunization against GRF reduced (P < .05) serum IGF-I (GRFi = 50 +/- 4 vs HSAi = 137 +/- 4 ng/mL) and BW (GRFi = 71 +/- 3 vs HSAi = 105 +/- 3 kg) and increased (P < .05) backfat depth (GRFi = .38 +/- .03 vs HSAi = .25 +/- .02 mm/kg). Age at puberty was similar in GRFi and HSAi gilts, but ovulation rate was lower (P < .05) after third estrus in GRFi (11.3 +/- .8) than in HSAi (13.8 +/- .8) gilts. Thus, GRFi at 92 or 35 d of age decreased serum ST, IGF-I, and BW in prepubertal gilts without altering age of puberty. However, GRFi at 35 d of age, but not 92 d of age, decreased ovulation rate. These results indicate that alterations in the somatotropic axis at 1 mo of age can influence reproductive development in pubertal gilts.     

Follicular growth and production of estrogen and progesterone after injection of gilts with human chorionic gonadotropin on day 12 of the estrous cycle

Twenty cyclic gilts were injected im with either saline (control) or 1,000 IU of human chorionic gonadotropin (hCG) on d 12 of the estrous cycle to determine the effects of hCG on follicular development and steroidogenesis. Blood was collected when gilts were sacrificed on d 13 or 16. Follicles were classified as medium (3 to 6 mm in diameter) or large (greater than 6 mm diameter), dissected from the ovary, measured and weighed. Pieces of follicle wall were incubated 3 h in Krebs Ringer bicarbonate buffer (KRB) on ice in an atmosphere of air or at 37 C in an atmosphere of 95% O2:5% CO2. Unconjugated estrogen and progesterone in blood plasma, follicular fluid and 10,000 X g supernatants of incubated follicular tissue homogenates were quantified by radioimmunoassay. On d 13 follicles on ovaries of control or hCG-injected gilts were less than or equal to 6 mm in diameter. On d 16, one of five control gilts had some large follicles, while all five hCG-treated gilts had large as well as medium follicles. On d 16 follicular fluid of large follicles from hCG-injected gilts contained twofold more estrogen and 40-fold more progesterone than medium follicles on the same ovaries. Tissue from large follicles of hCG-injected gilts produced more progesterone in vitro than did tissue from medium follicles (P less than .05), but estrogen production did not differ. On d 16 medium follicles from control or hCG-injected gilts were larger, contained more estrogen and less progesterone than those recovered on d 13 (P less than .01).(ABSTRACT TRUNCATED AT 250 WORDS)     

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)     

Changes in follicular endocrinology during final maturation of porcine oocytes

Thirty-one gilts were ovariectomized between 21 and 34 hr after the onset of estrus to compare changes in follicular endocrinology with stages of oocyte maturation. Oocytes were recovered from 6 to 8 mm follicles and classified by stage of meiosis. Remaining follicular fluid was assayed for steroids and dermatan sulfate. Amounts of prostaglandin F2 alpha (PGF2 alpha) and E2 (PGE2) were measured in intramural tissues. Coincident with germinal vesicle breakdown, the follicular content of all steroids except testosterone decreased (P less than .05). As oocytes approached metaphase II, the amount of progesterone within follicles increased (P less than .05), and estradiol continued to decrease (P less than .05). The pattern of dermatan sulfate content was biphasic and peaked at germinal vesicle breakdown and anaphase stages. Amounts of PGF2 alpha and PGE2 within intramural tissues increased (P less than .05) throughout oocyte maturation. Follicular atresia was evident during estrus; however, more (P less than .05) atretic follicles were recovered at germinal vesicle than metaphase II stages (20 vs 3%, respectively). Follicular development, within a gilt, was skewed (P less than .05) and classification of follicles by hormone content demonstrated that a majority were more mature than a minority of less mature follicles. These data suggest that follicular maturation and oocyte development are highly correlated in swine. Furthermore, partitioning the follicular variability by hour and stage of oocyte maturation allowed for more precise assessment of follicular endocrinology than previously reported.     

Effects of gonadotropin treatment on ovarian follicle growth and granulosal cell aromatase activity in prepuberal gilts

This experiment was conducted to compare the ability of USDA porcine FSH-B-1 (pFSH), USDA porcine LH-B-1 (pLH), and pregnant mare's serum gonadotropin (PMSG) to grow large follicles and induce granulosal cell aromatase activity in prepuberal gilts. Twenty-four gilts (164 d old) received one of four treatments by i.m. injection: 1) saline once, n = 8; 2) pFSH (8 micrograms/kg BW, nine times at 8-h intervals), n = 5; 3) pLH (2 micrograms/kg BW, nine times at 8-h intervals), n = 6; or 4) PMSG (15 IU/kg BW, once), n = 5. At slaughter, 72 h after the first injection, the ovaries to saline-treated gilts contained an average of 104 surface antral follicles 1 to 3 mm in diameter. compared to treatment with saline, pFSH increased (P less than .05) the number of follicles 46%, whereas pLH or PMSG decreased (P less than .05) the number by 70 and 84%, respectively. Compared with saline, treatment with PMSG or pLH induced growth of large follicles (7 to 9 mm) (10.8 and 4.8 follicles/gilt, respectively), increased plasma estrogen, increased granulosal cell aromatase activity, and decreased plasma FSH by 51 and 69%; treatment with pFSH had no significant effect on these traits. Results indicate that injected pFSH did not cause growth of large follicles or induce granulosal cell aromatase activity in prepuberal gilts. In contrast, LH initiated growth and increased granulosal cell aromatase activity in a small number of follicles and accelerated atresia among the remaining follicles.