PubMed Health⌕ Search

Biomedical subjects

J H Britt

Publications and source records attributed to J H Britt.

At least 19 recordsLinked to original sources

Effect of long-term treatment with recombinant bovine somatotropin and estradiol on hormone concentrations and ovulatory response of superovulated cattle.

The objective was to assess effects of long-term treatment with recombinant bovine somatotropin (bST) and estradiol-17beta (E2) on the number of follicles that ovulated in response to FSH. Non-lactating Holstein and Jersey cows (Trial 1, n=27) and Angus cows and heifers (Trial 2, n=35) received two ear implants of E2 and biweekly injections of bST in a 2 x 2 arrangement of treatments. Estradiol implants were removed 74.6 +/- 1.1 d after insertion and 18.1 +/- 0.9 d after the last biweekly injection of bST. Cows were stimulated with FSH-P beginning 2 d after removal of E2 implants, and PGF2alpha (PGF) was given on the third day of FSH treatment. Ovaries were collected to determine the number of CL at 1 to 2 wk after treatment with PGF. In Trial 2 only, cattle were inseminated at estrus and embryos were collected 6 to 8 d later. Implants of E2 increased (P < 0.01) serum E2 8-fold initially and E2 was still elevated 5-fold at removal of implants. Injections of bST increased (P < 0.01) serum growth hormone (GH) 15-fold and insulin-like growth factor-I (IGF-I) 3-fold. In Trial 1, number of CL was increased by the combination of bST+E2 (P < 0.01). In Trial 2, E2 increased the number of CL (P < 0.05), and bST increased the number of total ova and transferable embryos (P < 0.01). We conclude that long-term treatment with bST and E2 may interact to enhance follicular development and ovulatory response to FSH.

Animals↗

Ovarian follicular development following administration of progesterone or aspiration of ovarian follicles in Holstein cows.

The objective of this study was to compare the effects of administration of a single injection of progesterone (P4) and follicle aspiration on Day 7 of the estrous cycle on the timing and synchrony of follicular wave emergence, time of ovulation, and concentrations of P4, estradiol and FSH in Holstein cows. Twenty cows were assigned to 4 groups (n=5 cows per group) in a 2 by 2 factorial arrangement. Cows were treated on Day 7 (Day 0 = estrus) of the estrous cycle with either sham follicular aspiration and an oil vehicle administered intramuscularly (control), aspiration of ovarian follicles (aspiration), 200 mg of P4 im, or aspiration and 200 mg of P4 im (aspiration + P4). On Day 11, PGF(2alpha)(25mg) was administered to all groups. Synchrony of ovulation was less variable in each of the treatment groups compared with the control group (P<0.05), whereas ovulation was delayed in cows in the P4 group (P<0.05). Day of follicular wave emergence was delayed in the cows of the P4 group compared with cows in the aspiration and aspiration + P4 groups (P<0.01), whereas variability in wave emergence was less among both groups of aspirated cows compared with the cows in the control group (P<0.01). More follicles 4 to 7 mm in diameter were detected in the 2 aspiration groups compared with the cows in the control and P4 group (P<0.05). No difference was detected among groups in the maximum concentration of FSH associated with follicular wave emergence. We conclude that both the administration of P4 and the aspiration of follicles on Day 7 of the estrous cycle improves the synchrony of ovulation when luteolysis is induced on Day 11 and results in similar concentrations of FSH at the time of follicular wave emergence, but the timing of wave emergence and the number of follicles post-emergence differ.

Animals↗

Alteration of activation, growth, and atresia of bovine preantral follicles by long-term treatment of cows with estradiol and recombinant bovine somatotropin.

The hypothesis was that long-term treatment of cattle with estradiol (E(2)) and bovine somatotropin (bST) would alter the earliest stages of folliculogenesis. Nonlactating Holstein cows (n = 26) were treated in a 2 x 2 arrangement with E(2) (2 x 24 mg implants, 67.1 +/- 1.4 days) and bST (Posilac, 63.6 +/- 1.5 days). At Day 67 +/- 1.3, one ovary was removed for morphometric and immunohistochemical analysis. For each ovary, 388 +/- 38 microscopic fields (2 x 2 mm) were examined and follicles within each field were classified by histological stage. Fields that contained no follicles were classified as empty. Empty fields (n = 100 per ovary) were further classified as containing no evidence of follicles or containing atretic remnants of follicles. Approximately 30 4-microm sections per ovary were stained for proliferating cell nuclear antigen (PCNA), and 150 fields per ovary were evaluated. Additional sections (n = 10 per ovary) were assessed immunohistochemically for apoptosis, and fluorescence intensity was determined for each follicle. Treatment with bST significantly decreased percentage of empty fields containing atretic remnants. Treatment with E(2) induced activation of follicles as shown by a decrease in percentage of primordial follicles and an increase in percentage of primary follicles as determined by PCNA staining. At the primary follicle stage the combination of bST + E(2) decreased apoptosis as shown by decreased fluorescence intensity. Thus, E(2) induced activation of follicles, bST enhanced survival, and the combination lowered atresia.

Animals↗

The effect of early postnatal treatment with a gonadotropin-releasing hormone agonist on the developmental profiles of testicular steroid hormones in the intact male pig.

Three studies examined the effects of early postnatal treatment with a GnRH agonist on plasma concentrations of testosterone, dehydroepian-drosterone sulfate, 16-androstene steroids in fat and salivary glands, androstenone in fat and plasma, and testicular development of intact male pigs. The first study involved 45 7-d-old pigs assigned to three treatment groups: 1) boars administered 100 microg/kg of Lupron depot, 2) boars administered 200 microg/kg of Lupron depot, and 3) control boars receiving a saline carrier. The second study involved 20 7-d-old pigs assigned to two treatments: daily injection of 200 microL of 0.5 mg/mL Lupron from d 7 to 35 and controls treated with saline. The third study involved a total of 100 animals assigned to 10 groups of 10 based on their age at slaughter. These groups were subdivided into one of two treatments: 1) boars injected with 200 microL of 0.5 mg/mL of Lupron from d 3 to 35 and 2) control boars injected with saline. Testicular steroid hormone concentrations in plasma decreased (P < 0.01) within 7 d of GnRH agonist treatment. Following cessation of treatment, steroid levels increased to control levels and remained constant until the final rise at 5 mo. Plasma testosterone levels in the 100 microg/kg depot treatment group were higher (P < 0.05) than that of the 200 microg/kg and control group at 164 d of age. There were no differences between treatments (P > 0.05) in testicular steroid hormone levels at the end of study 2 or 3. There were no differences (P > 0.05) in concentrations of 16-androstene steroids in salivary glands between any of the treatment groups at market weight in studies 1 and 2. Fat androstenone levels measured in the third study ranged between 0.6 microg/g and 4.2 microg/g at 7 to 28 d of age. Treatment with GnRH agonist decreased plasma steroid levels and testicular development; however, by d 60 testicular size and weight were at control levels and remained similar until 180 d of age. The results of these studies indicate that daily administration of a GnRH agonist significantly decreased testicular development and steroidogenesis only during treatment, but testis growth and steroidogenesis had returned to control levels by 60 d of age in male pigs. Suppression of the early postnatal rise in testicular steroid hormones did not affect growth performance or steroid hormone levels at 5 to 6 mo of age.

Adipose Tissue↗

In vivo oxytocin release from microdialyzed bovine corpora lutea during spontaneous and prostaglandin-induced regression.

The release of luteal oxytocin during spontaneous and prostaglandin-induced luteolysis was investigated in cows. A continuous-flow microdialysis system was used in 11 cows to collect dialysates of the luteal extracellular space between Days 12 and 24 postestrus. Seven cows were untreated and were expected to exhibit spontaneous luteolysis during sampling, whereas 4 cows received prostaglandin F(2alpha) (PGF(2alpha)) systemically between Days 13 and 15 to induce luteolysis during sampling. Oxytocin was detectable in the dialysate of all cows before Day 16 postestrus and occurred as 2 or 3 discrete pulses per 12-h sampling period. For non-PGF(2alpha)-treated cows, dialysate oxytocin content began to decline spontaneously on Day 15 postestrus and was undetectable by Day 17 postestrus. Oxytocin decay curves preceded onset of serum progesterone decline by at least 72 h and were not related temporally with onset of progesterone decline within cow. Exogenous PGF(2alpha) (25 mg, i.m.) produced a 10-fold increase in dialysate oxytocin within 1 h (1.9 +/- 0.3 pg/ml to 20.8 +/- 3.0 pg/ml; P < 0. 01). Dialysate oxytocin then declined to pretreatment concentrations within 2 h and was undetectable within 8 h posttreatment. A second PGF(2alpha) injection given 20 h after the first did not result in a measurable increase in dialysate oxytocin, probably because luteolysis was underway. Although robust luteal oxytocin release was observed after treatment with a pharmacological dose of PGF(2alpha), the lack of detectable oxytocin secretion during spontaneous luteolysis suggests that the contribution of luteal oxytocin in the cow may be less than that proposed for the ewe.

Animals↗

Evaluation of numbers of microscopic and macroscopic follicles in cattle selected for twinning.

We hypothesized that the number of microscopic follicles present in the ovaries of cattle selected for twin births (Twinner) would be greater than in the ovaries of contemporary Controls. Ovaries were collected from seven Control and seven Twinner cows at slaughter. The number of Small (1 to 3.9 mm), Medium (4 to 7.9), and Large (> 8 mm) surface follicles was counted and one ovary was fixed for histological evaluation. Fifty to sixty consecutive 6-microm slices were taken from a piece of cortical tissue, approximately 1 cm x 1 cm in area, located between the surface follicles. Microscopic follicles were classified as primordial (oocyte surrounded by a single layer of squamous pregranulosa cells), primary (oocyte surrounded by a single layer of one or more cuboidal granulosa cells), secondary (oocyte surrounded by two or more layers of granulosa cells), or tertiary (oocyte surrounded by multiple layers of granulosa cells with initiation of antrum formation to < or = 1 mm in diameter). The total number of follicles was counted in 200 fields (2 mm x 2 mm) per ovary. A field containing no follicles was classified as empty. There were significantly more secondary follicles in Twinner compared with Control ovaries (12.9 vs 6.3; P < .05). Twinners also tended to have more small surface follicles (35.4 vs 49.0; P < 0.1). We conclude that ovaries of Control and Twinner cows do not differ in the number of primordial follicles or in the number of follicles activated into the growing pool; however, Twinner cows are able to maintain more growing follicles at the secondary and subsequent stages of development.

Animals↗

Influence of vitamin A injection before mating on oocyte development, follicular hormones, and ovulation in gilts fed high-energy diets.

Previous research revealed that treatment with vitamin A approximately 5 d before ovulation may increase litter size in weaned sows and improve embryonal survival in gilts fed high-energy diets that reduced embryonal survival. For the current study, the hypothesis was that administration of vitamin A before ovulation would alter development of follicles and oocytes in a way favorable to enhanced embryonal survival. (Landrace x Large White) x (Duroc x Hampshire) gilts (n = 44) were fed 11.0 Mcal ME x gilt(-1) x d(-1) beginning 7 d after second estrus and given (i.m.) corn oil or 1 x 10(6) IU of vitamin A (retinyl palmitate) on d 15 after second estrus. Gilts were checked for estrus every 4 h, mated naturally at third estrus, and assigned randomly to undergo midventral laparotomy beginning at 24 to 28, 28 to 32, 32 to 36, or 36 to 40 h after onset of third estrus. At laparotomy, ovulated oocytes and early-stage embryos were recovered from oviducts, and ovaries were removed for aspiration of oocytes and granulosa cells from unovulated follicles. Oocytes and embryos were stained for assessment of stage of development. Granulosa cells were cultured to assess their ability to secrete progesterone. Follicular fluid was assayed for progesterone, estradiol-17beta, IGF-I, and PGF2alpha. Treatment with vitamin A altered development of oocytes and embryos by decreasing the percentage at the germinal vesicle stage and increasing the percentage at advanced stages. Mean stage of development was increased by vitamin A, but variation in stage was decreased. Among follicles matched by meiotic stage of oocyte, follicular fluid concentrations of progesterone, IGF-I, and PGF2alpha were greater in vitamin A-treated gilts than in controls, but treatment with vitamin A in vivo did not affect LH-stimulated or unstimulated secretion of progesterone by granulosa cells in vitro. These data provide evidence that vitamin A may influence embryonic development by advancing resumption of meiosis and altering follicular hormonal environment during follicle maturation.

Animal Feed↗

Estimates of pregnancy outcomes based on selection of bovine embryos produced in vivo or in vitro.

The objective of this study was to estimate the degree of variation among experienced evaluators selecting in vivo- or in vitro-produced embryos for transfer and to determine how this affects both the proportion of recipients becoming pregnant after transfer, and the number of embryo transfers required per pregnancy. Data from 6 experienced evaluators who graded Day 7 embryos produced either in vivo (n = 15) or in vitro (n = 15) were used to estimate these effects. The evaluators viewed video recorded images of the embryos and classified each embryo for stage of development and quality grade (1 = excellent, 2 = good, 3 = fair, 4 = degenerated and nontransferable). The statistical model considered transfer of embryos of the following individual or combined grades: Grade 1 only, Grade 2 only, Grade 3 only, Grades 1 and 2, Grades 2 and 3, and Grades 1, 2 and 3. Probabilities of pregnancy after embryo transfer were based on pregnancy rates of recipients at the facility of 1 of the 6 evaluators where the percentages of heifers pregnant after the transfer of Grade 1, 2 and 3 embryos, by embryo source, were 76, 65 and 54% (in vivo), and 59, 45 and 30% (in vitro). Within most grades, the proportion of embryos selected for transfer differed (P < 0.05) among the 6 evaluators. Although no significant differences (P > 0.10) were found among evaluators in the proportion of recipients pregnant after transfer within any embryo grade, there was substantial variation among evaluators in the proportion of recipients becoming pregnant, especially for embryos produced in vitro. Estimated percentages of heifers becoming pregnant for embryos classified as Grade 1, 2 or 3 were 66 to 76, 62 to 69, and 54 to 60%, respectively, for in vivo-produced embryos; and, 39 to 59, 15 to 45, and 24 to 32%, respectively, for in vitro-produced embryos. Approximately twice as many transfers were needed per pregnancy for embryos produced in vitro as for those produced in vivo regardless of the grade.

Animals↗

Superovulatory response of one ovary is related to the micro- and macroscopic population of follicles in the contralateral ovary of the Cow.

We hypothesized that the ovulatory response of one ovary to FSH would be related positively to the size of the primordial and growing pools of follicles in the other ovary. Nonlactating cows (n = 26) were unilaterally ovariectomized and 2 days later were superovulated. The superovulatory response was classified as Low (< 5 corpora lutea [CL]), Medium (5-14 CL), or High (> 14 CL). Surface follicles on the ovary removed before superovulation were classified as small (1-3 mm), medium (3-7 mm), or large ( > 7 mm), and the ovary was then fixed and serially sectioned. Follicles </= 1 mm in diameter in 388 +/- 38 fields (2 x 2 mm) per cow were classified as primordial, primary, secondary, or tertiary. By classification, Suboptimal ovaries contained < 100 follicles </= 1 mm and Optimal ovaries contained > 250 follicles </= 1 mm. Number of CL was correlated positively with total number of primordial, tertiary, and medium surface follicles. Number of Empty fields (2 x 2-mm fields containing no follicles) was correlated negatively with superovulatory response and number of primordial follicles. Number of CL was related to number of tertiary follicles in a positive linear manner and the number of medium follicles in a positive quadratic manner (r 2 = 0.66). Numbers of primordial, tertiary, small surface follicles, medium surface follicles, and total surface follicles were lower (p </= 0.06) in the Low superovulatory response group than in the Medium or High group. Suboptimal ovaries had fewer small surface follicles and fewer CL than Optimal ovaries (p < 0.05). We conclude that superovulatory response in cattle is related positively to the pools of primordial and growing follicles in the bovine ovary.

Animals↗

Technical note: use of slow-release estradiol and prostaglandin F2alpha to induce pseudopregnancy and control estrus in gilts.

We determined whether a single injection of slow-release estradiol-17beta (SRE2) would induce pseudopregnancy in gilts and whether PGF2alpha would regress the corpora lutea (CL) of pseudopregnancy. Crossbred gilts (n = 40) were induced to ovulate by treatment with 400 IU of hCG + 200 IU of eCG (PG600, Intervet, Millsboro, DE) given at 180 d of age (d = 0). On d 14, gilts were injected i.m. with one of five doses (n = 8 gilts/dose) of SRE2 (0, 12.5, 25, 50, or 100 mg). Blood samples were collected before SRE2 and twice weekly until d 73 to monitor serum progesterone (P4) and estradiol (E2). On d 59, gilts received (i.m.) 10 mg of PGF2alpha (Lutalyse, Pharmacia Upjohn, Kalamazoo, MI) and were checked for estrus for 7 d. On d 62, mammary development was scored (0 = no development; 1 = some development; 2 = teat and gland development) by a neutral observer. Treatment with SRE2 increased (P < .05) peak E2 concentrations, duration of luteal function, and mammary gland score. There were no differences (chi-square, P > .05) among doses of SRE2 in the percentage of pseudopregnant gilts that showed luteolysis after PGF2alpha. We conclude that a single injection of SRE2 can induce pseudopregnancy and that the CL can be regressed with PGF2alpha, providing a simple method for controlling estrus in gilts.

Animals↗

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.

Animals↗

Control of the weaning-to-estrus interval in sows using gonadotropins and prostaglandins during lactation.

Two experiments were conducted to examine the effectiveness of various strategies using gonadotropins to induce ovulation during lactation as a means of controlling the weaning-to-estrus interval in sows. The objective of Exp. 1 was to examine the efficacy of various gonadotropin regimens for induction of ovulation during lactation. Primiparous (n = 60) and multiparous (n = 83) crossbred sows were assigned, before farrowing, to one of four treatments: no injection (control); 1,000 IU hCG on d 0 (hCG-0; d 0 = day of farrowing); P.G. 600 + 1,000 IU hCG 4 and 7 d after farrowing, respectively (hCG-7); or P.G. 600 + 1,000 IU hCG 11 and 14 d after farrowing, respectively (hCG-14). Sows were weaned on 18 +/- 2 d after farrowing and monitored daily for estrus via exposure to mature boars. The criterion for determining the induction of ovulation was a sustained increase in serum progesterone concentrations above 4.0 ng/mL. The most consistent response to exogenous gonadotropins was on d 0, with an 80% response in primiparous sows (12/15) and a 71% response in multiparous sows (15/21). Weaning-to-estrus intervals for multiparous sows were longer (P = .05) for hCG-14 and hCG-7 than for control and hCG-0 sows. Weaning-to-estrus intervals for primiparous sows were longer (P = .05) for the hCG-14 than for the hCG-0 treatment. The objective of Exp. 2 was to ascertain the effects of postpartum treatment with hCG (1,000 IU) on d 0 and PGF2alpha (10 mg) at d 14 on the weaning-to-estrus interval in multiparous sows weaned at d 14 after birth. Before farrowing, sows (n = 60) were randomly assigned to one of four treatments: positive control, weaning at d 21; negative control, weaning at d 14; hCG within 24 h after farrowing, weaning at d 14; or hCG within 24 h after farrowing and PGF2alpha at weaning, weaning at d 14. Weaning-to-estrus intervals were longer (P = .05) in sows receiving PGF2alpha than in the other treatments. Results indicate that it is possible to induce ovulation immediately after farrowing, using a single injection of hCG, and this strategy can be used to uncouple weaning from the resumption of reproductive activity. However, the administration of PGF2alpha at 14 d after farrowing did not consistently cause regression of the induced corpora lutea.

Animals↗

Role of gonadotropin-releasing hormone pulse frequency in differential regulation of gonadotropins in the gilt.

We tested the hypothesis that different GnRH pulse frequencies will affect serum LH and FSH differently. Ovariectomized gilts (n = 6), immunized against GnRH, were given 200-ng pulses of GnRH agonist (GnRH-A) every 180 min for 3 days (pretreatment), followed by GnRH-A pulses every 30, 60, or 180 min for 3 days (treatment) in a Latin rectangle design. Mean gonadotropin concentrations did not change over time when GnRH pulses were administered every 180 min. Initiation of high GnRH-A pulse frequency (30 min) caused a robust increase in serum LH to 265% of the pretreatment level (p < or = 0.007) and a more moderate increase in serum FSH to 127% of pretreatment level (p < or = 0.02). After 66 h of frequent pulsing, desensitization had occurred and serum LH concentrations were similar to pretreatment concentrations, but serum FSH had decreased to 53% of pretreatment levels (p < or = 0.0008). After 72 h of treatment, 5 micrograms GnRH-A was infused to estimate residual releasable pools of LH and FSH, and the amounts of LH and FSH released were negatively correlated with GnRH-A pulse frequency. The results of this study imply that the LH surge is terminated because the pituitary gland becomes incapable of responding to an otherwise adequate stimulus, and not because of exhaustion of releasable LH pools. Our results confirm that in the pig the response to altered GnRH-A pulse frequency differs between LH and FSH. High GnRH pulse frequency is more effective in acutely releasing LH than FSH. Low pulse frequency of GnRH supports FSH synthesis and release, but is not as effective in increasing LH concentrations, while high GnRH pulse frequency inhibits FSH synthesis and release.

Activity Cycles↗

Evidence that injection of vitamin A before mating may improve embryo survival in gilts fed normal or high-energy diets.

The hypothesis was that administration of vitamin A before ovulation would improve embryo survival in gilts fed a high-energy diet intentionally to reduce embryo survival. Forty crossbred ([Landrace x Large White] x [Duroc x Hampshire]) gilts were fed control (5.5 Mcal ME/d) or high-energy (11.0 Mcal ME/d) diets from 7 d after second estrus until 11 to 12 d after third estrus. Gilts in each dietary group received (i.m.) corn oil or retinyl palmitate (1 x 10(6) IU, vitamin A) on d 15 after second estrus and were mated at third estrus. Blood for determination of progesterone and estradiol was collected twice daily. The uterus and ovaries were removed on d 11 or 12 after third estrus for assessment of number of CL, and number, size and aromatase activity of embryos. Neither diet nor vitamin treatment affected number of CL. The high-energy diet exerted a negative effect on number of embryos (P = .09) and embryo survival (P = .07), whereas vitamin A exerted a positive effect on number of embryos (P = .07) and embryo survival (P = .08). The high-energy diet increased variation in embryo diameter, whereas vitamin A reduced variation in diameter and increased average diameter. Neither diet nor vitamin treatment influenced aromatase activity of embryos. Diet and vitamin treatment interacted with day to influence serum progesterone, but not estradiol. Injecting vitamin A before estrus restored embryo survival to normal levels in gilts fed high-energy diets, and this may be attributable to decreased variation in size of embryos.

Animals↗

Localization of laminin proteins during early follicular development in pig and rabbit ovaries.

Laminin, a major component of the basal lamina, is known to be important in the differentiation of epithelial cells. The outer granulosa cell layer of ovarian follicles is attached to a basal lamina surrounding the follicle and it has been demonstrated that proteins of the basal lamina can alter the steroidogenic capacity and cytoskeletal composition of mature granulosa cells. The present studies were carried out to evaluate the developmental expression of laminin proteins in the basal lamina of ovarian follicles during early stages of granulosa cell differentiation. Ovaries from sexually immature pigs (0-20 weeks of age) and rabbits (0-12 weeks of age) were used for this study because formation of primordial follicles and initiation of follicular growth occur partially or completely postnatally. Specific antibodies were made against laminin beta 1/gamma 1 chains using protein purified by high resolution two-dimensional polyacrylmide gel electrophoresis. These antibodies were used in immunohistochemical localization studies to demonstrate that laminin is associated with the basal lamina surrounding egg clusters and their connections to the ovarian surface epithelium and ovarian rete during early ovarian development. In addition, laminin was observed to associate with a continuous matrix that surrounds forming primordial follicles as they are isolated from the egg clusters. Laminin is localized in the basal lamina of primordial, primary, secondary, and tertiary follicles of both pig and rabbit ovaries. Laminin proteins are not only expressed throughout early follicular development in pigs and rabbits, but are also found surrounding the germ cell compartments prior to initiation of meiosis and formation of primordial follicles. These results demonstrate that laminin proteins are deposited in the basal lamina well before granulosa cells undergo morphological differentiation. This pattern of expression suggests that laminin proteins alone do not control changes in granulosa cell morphology during early development but may be required to maintain cell lineage commitment.

Animals↗

Does increased PGF2 alpha receptor concentration mediate PGF2 alpha-induced luteolysis during early diestrus in the pig?

The mechanism by which multiple injections of PGF2 alpha result in premature luteolysis in pigs is unknown. In the present study we evaluated whether PGF2 alpha receptor concentrations on large luteal cells changed when gilts were injected IM with 12.5 mg of PGF2 alpha every 12 hours from the morning of day 5 of an estrous cycle (estrus = day 0) until ovariectomy on day 6, 7, 8, or 9. Luteal PGF2 alpha receptor concentrations remained constant from day 6 through 9 in the PGF2 alpha-treated group, but increased linearly (P > 0.05) in control gilts from day 6 to 9. Receptor affinity for PGF2 alpha did not change throughout the study in either PGF2 alpha-treated or control gilts. Luteal progesterone concentrations were significantly lower in PGF2 alpha-treated gilts than in control gilts only on day 9. Histological examination of luteal tissue obtained from PGF2 alpha-treated gilts revealed definite evidence of luteolysis by day 8. We conclude that PGF2 alpha-induced premature luteolysis is not mediated by an increase in luteal PGF2 alpha receptor concentrations and, based on luteal progesterone concentrations and histological features, that the PGF2 alpha-based protocol used to shorten the estrous cycle is accompanied by premature functional and structural luteal regression.

Animals↗

Agreement among evaluators of bovine embryos produced in vivo or in vitro.

Six experienced individuals evaluated 40 embryos on videotape for stage of development and quality grade. These 40 observations comprised 15 embryos produced in vivo, 15 embryos produced in vitro, and 10 embryos that were repeated throughout the videotape. Embryos produced in vivo were recovered from uterine flushings of superovulated heifers 7 d after estrus, and embryos produced in vitro were harvested 7 d after insemination of in vitro-matured oocytes. Embryos of various stages (morulae, blastocysts, or degenerated) and quality grades (1 = excellent, 2 = good, 3 = fair, 4 = degenerated) were recorded on videotape for evaluation. After video microscopy, the embryos were stained and the number of nuclei per embryo was counted. Six evaluators reviewed the videotape and the percentage of agreement and kappa (k; agreement beyond chance) among evaluators were determined for classifications of stage and grade. Consistency of each evaluator's responses was estimated using the 10 repeated embryos. Agreement within evaluators was higher for stage of embryo development (89.2%) than quality grade (68.5%). Agreement among evaluators for stage was slightly higher with embryos produced in vivo (85.0%, k = 0.74) than in vitro (72.3%, k = 0.48). Agreement among evaluators for grade was similar with embryos from in vivo (61.0%, k = 0.46) and in vitro (57.7%, k = 0.42) production. For both sources of embryos, agreement was substantially better for Grades 1 and 4 than for Grades 2 and 3. The results of this study suggest that good to excellent agreement exists for classifying Day 7 bovine embryos by stage and by extremes of quality grade (Grades 1 and 4) but not by degree of abnormal morphology (Grades 2 and 3). Simple grading criteria of Grade 1 (highest quality), Grade 2 (morphologic defects), and Grade 3 (degenerated) maximized agreement among evaluators.

Journal Article↗

Concentrations of tumor necrosis factor alpha and progesterone within the bovine corpus luteum sampled by continuous-flow microdialysis during luteolysis in vivo.

Corpora lutea (CL) of mature, nonlactating Holstein cows (n = 12) each received surgical implants of a microdialysis system between Days 12 and 16 of an estrous cycle (estrus = Day 0). Fractions from dialyzed CL were collected every 30 min for 12 h each day for 7-9 days after surgery. Concurrent sampling of jugular venous blood allowed comparison between luteal dialysate and peripheral serum hormone concentrations. Six cows received no treatment, while 4 cows received prostaglandin F2 alpha (PGF; 25 mg, i.m.) and 2 cows received hCG (5000 IU, i.m.) within 48 h after surgery. Serum progesterone (P4) declined to < 1 ng/ml by Day 20 +/- 0.5 in 4 of 6 nontreated cows and by Day 15 +/- 0.6 in all 4 PGF-treated cows. Both of the hCG-treated cows had increased serum and dialysate P4 after treatment and did not show evidence of luteolysis by the end of their sampling period (Days 22 and 18), and 2 of 6 nontreated cows did not show evidence of luteolysis by the end of their sampling period (Days 22 and 24). All 8 cows undergoing spontaneous or induced luteolysis showed a significant rise (p < 0.01) in dialysate (but not serum) tumor necrosis factor alpha (TNF) after P4 began to decline. From the start of the decline in dialysate P4, intervals to first detectable TNF and to peak TNF, and duration of TNF in the dialysate were 25.8 +/- 8 h, 38.3 +/- 8 h, and 42.3 +/- 7 h, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗