W F Pope

The Ohio State University, Columbus, OH, United States

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Publications (51)116.6 Total impact

  • Horacio Cárdenas, William F Pope
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    ABSTRACT: Determination of the specific roles of the estrogen receptor (ER) forms in reproductive processes of different species remains incomplete. In the present experiment, cellular localization and changes in relative amounts of the ERα and ERβ in late developing ovarian follicles, oviduct, and uterus were determined during the follicular phase of the estrous cycle in sheep. Ewes in mid-luteal phase were treated with prostaglandin F(2α) (PG) to induce luteolysis and control the onset of the follicular phase. The oviducts, uterus, and the ovaries were collected at 0 (ewes not treated with PG), 4, 18, and 36 h after PG treatment (early, mid, and late follicular phase, respectively) and processed to evaluate the ERs using immunohistochemical (IHC) procedures. The ERα was localized to nuclei of granulosa cells of late developing follicles and most cells of the oviduct and uterus. The ERβ was detected only in ovarian follicles using two antibodies directed to different regions of the ERβ. Western immunoblotting demonstrated that the antibody directed against the N-terminal region of the ERβ detected one isoform (approximately 53 kDa) whereas the antibody directed against the C-terminus detected two ERβ isoforms (approximately 53 kDa and 59 kDa). Western and IHC results combined indicated presence of the 59 kDa ERβ in granulosa cells and the 53 kDa ERβ in both granulosa and theca cells. Relative amounts (immunostaining intensity) of the ERα increased (P<.05) in granulosa cells of preovulatory follicles and in the isthmian muscularis of the oviduct at the late follicular phase. Amounts of the ERα in the mucosal epithelium of the oviductal regions (isthmus, ampulla, and infundibulum), and in various uterine cell types (glandular and luminal epithelia, endometrial stromal cells, and myometrium) did not change (P>.05) throughout the follicular phase. A major increase (four-fold) in expression of the 53 kDa ERβ in the theca and a less pronounced increase in the granulosa occurred at the late follicular phase. The ERα is broadly expressed in reproductive organs of sheep and is upregulated only in few cell types during the late follicular phase. Immunoreactive ERβ was detected only in the ovary. Important estrogen actions in theca cells during preovulatory follicular development likely occur in association with a major increase in expression of an ERβ isoform.
    Animal reproduction science 03/2012; 131(3-4):143-52. · 1.56 Impact Factor
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    ABSTRACT: To examine how androgens affect endocrine events associated with increased ovulation rate, gilts were injected with androgen receptor agonists, an antagonist, or a combination of both. Blood samples were collected hourly from Day 13 to estrus (Day 0 = onset of estrus) coincident with gilts (n = 6 per treatment) receiving daily treatments of vehicle (corn oil), 10 mg of testosterone, 10 mg of 5 alpha-dihydrotestosterone (dihydrotestosterone), 1.5 g of flutamide (an androgen receptor antagonist), testosterone plus flutamide, or dihydrotestosterone plus flutamide. Treatment of gilts with testosterone or dihydrotestosterone alone increased (P < 0.05) concentrations of FSH in serum, and these effects were blocked by cotreatment with flutamide. Estradiol-17beta and androstenedione concentrations in serum were increased (P < 0.05) at 2 h after injection of testosterone or testosterone plus flutamide but not after dihydrotestosterone treatment, probably because of the role of testosterone as a substrate for estradiol-17beta and androstenedione synthesis. There were no effects of the six treatments on serum concentrations of progesterone during luteolysis, but treating gilts with testosterone shortened (P < 0.05) the proestrous period. Total embryonic loss by Day 11 in gilts treated with dihydrotestosterone was reversed when gilts were cotreated with dihydrotestosterone plus flutamide. Results of this experiment indicated that androgen actions both increased FSH secretion and reduced embryonic survival by a mechanism(s) dependent on the androgen receptor.
    Biology of Reproduction 08/2008; 79(6):1148-52. · 4.03 Impact Factor
  • H Cárdenas, E Jiménez, W F Pope
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    ABSTRACT: The present experiments were conducted to determine androgenic effects on numbers, health, and amounts of gonadotropin receptor mRNA in late developing follicles of gilts. Gilts (n=5 per group) received daily injections of one of the following treatments on days 13-16 or days 13-18 of the estrous cycle: corn oil, 5alpha-dihydrotestosterone (DHT, 10 mg), flutamide (1.5 g, an androgen receptor inhibitor), DHT (10 mg) plus flutamide (1.5 g), testosterone (10 mg), and testosterone (10 mg) plus flutamide (1.5 g). Ovarian follicles > or =5 mm in diameter were evaluated on day 17 or 19, 24 h after receiving the last treatment dose. Follicles were classified as healthy (H), moderately atretic (MA), or very atretic (VA). Treatment with DHT increased (P<0.05) the numbers of H follicles relative to control gilts on days 17 and 19. DHT administration from days 13 to 16 diminished (P<0.05) the amounts of LH receptor (LHR) mRNA in H follicles from day 17 (relative amounts: 1.45+/-0.33 and 2.72+/-0.33 for DHT- and vehicle-treated gilts respectively). The effects of DHT on numbers of H follicles and LHR mRNA were not observed in gilts receiving DHT plus flutamide. Androgens did not influence numbers of MA, VA, and total follicles, or follicular estradiol-17beta concentrations and amounts of FSHR mRNA. Treating gilts with DHT during follicular recruitment and selection did not induce changes in the numbers of total follicles > or =5 mm, but rather increased the numbers of healthy follicles in this follicular population in association with decreased amounts of LHR mRNA.
    Reproduction 03/2008; 135(3):343-50. · 3.56 Impact Factor
  • W F Pope, H Cárdenas
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    ABSTRACT: Androgens have potential actions in almost all the organs of males and females. In females, most organs contain some tissues with cells that have androgen receptors. Androgens can regulate cellular functions by binding to androgen receptors or be converted to other hormones. For example, testosterone can bind to the androgen receptor or be aromatised to oestradiol. Treating animals with testosterone, therefore, might elicit some androgenic and oestrogenic effects. Alternatively, testosterone can be converted to other androgens, which in turn, have more or less affinity with the androgen receptor and these new metabolites may or may not be aromatised to oestrogens. This review will highlight the roles of androgens in female mammals other than those as a substrate for oestrogen, with particular emphasis on the actions of the androgen receptors in uteri and ovaries of pigs. Utilising small dosages of an androgen receptor agonist, DHT (5alpha-dihydrotestosterone) we have observed that some uterine functions were inhibited while ovarian follicular development was augmented. These inhibitory and stimulatory effects of androgen therapy on reproductive organs can potentially be balanced to enhance ovulation rate and litter size in gilts and sows. Perhaps after future experimentation, new uses of androgens or anti-androgens could improve additional aspects of sow performance not presently under consideration.
    Society of Reproduction and Fertility supplement 02/2006; 62:55-67.
  • Horacio Cárdenas, William F Pope
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    ABSTRACT: 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.
    Domestic Animal Endocrinology 11/2005; 29(3):523-33. · 2.38 Impact Factor
  • Horacio Cárdenas, Todd M Wiley, William F Pope
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    ABSTRACT: Effects of prostaglandin F(2alpha) (PGF(2alpha)), administered during the mid-luteal phase of the estrous cycle, were examined in ewes exhibiting estrous cycles classified as short (< or =16.5 days, short-cycle ewes, n = 10) or long (> or =18 days, long-cycle ewes, n = 9) based on the durations of two estrous cycles (cycles -2 and -1) before treatment. The ewes received (i.m.) 20mg of PGF(2alpha) on day 10 of the third estrous cycle (cycle 0) followed, 36 h later, by 25 microg of gonadotropin releasing hormone (GnRH) to time the events of ovulation. Duration of subsequent estrous cycles +1 and +2 were recorded, and then the ewes were treated with the same combination of PGF(2alpha) and GnRH beginning on day 10 of estrous cycle +3. Ovaries were recovered 6h after GnRH administration to assess development of pre-ovulatory follicles. The proportion of ewes that exhibited estrus after PGF(2alpha) and GnRH treatment on cycle 0 was not different (P > 0.05) between short- and long-cycle ewes. Onset of estrus occurred sooner (P < 0.05) after PGF(2alpha) injection in short-cycle ewes than in long-cycle ewes (1.9 +/- 0.1 days and 2.3 +/- 0.1 days, duration of cycle 0 was 11.9 and 12.3 days, respectively). Duration of estrous cycle +1 was 1.2 days longer (P < 0.01) than cycle -1 in short-cycle ewes. However, duration of estrous cycle +1 did not change (P > 0.05) after PGF(2alpha) and GnRH administration in ewes having long cycles. Pre-ovulatory follicles did not differ (P > 0.05) in numbers, diameter, layers of granulosa cells nor concentrations of progesterone and estradiol-17beta in follicular fluid between short- and long-cycle ewes after PGF(2alpha) and GnRH treatment. In conclusion, ewes having short or long estrous cycles responded differently to PGF(2alpha) and GnRH treatment with respect to the interval to onset of estrus and duration of the subsequent estrous cycle.
    Theriogenology 07/2004; 62(1-2):123-9. · 2.08 Impact Factor
  • Horacio Cárdenas, William F Pope
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    ABSTRACT: Androgens are known to attenuate some effects of estradiol-17beta (E) in the uterus. The objectives of the present experiment were to determine effects of 5alpha-dihydrotestosterone (DHT) on estrogenic actions in the pig uterus and its associations with changes in expression of the estrogen receptor (ER) alpha and ERbeta. Postpubertal gilts (120-130 kg of body weight; n = 16) were ovariectomized, and 3-4 weeks later received once-a-day injections (i.m.) of one of the following treatments during four consecutive days: 1) vehicle (corn oil), 2) E (250 microg), 3) E (250 microg) plus 1 mg DHT, or 4) E (250 microg) plus 10 mg DHT. Uterine tissues were collected 24 h after the last treatment. Gilts receiving E or E plus 1 mg DHT had greater uterine wet weight, uterine horn diameter, luminal epithelium thickness, and endometrial gland diameter compared with gilts treated with vehicle or E plus 10 mg DHT. Gilts receiving E or E plus 1 mg DHT were not different in these characteristics. Relative amounts of mRNAs in the endometrium for the cell proliferation marker histone H2a and the E-inducible protein complement component C3 increased in gilts treated with E compared with gilts treated with vehicle. E-induced increases in histone H2a and C3 mRNAs were not altered by cotreatment with E plus 1 mg DHT but were inhibited by E plus 10 mg DHT. Androgen receptor (AR) mRNA in the endometrium increased by treatment with E. Cotreatment of gilts with E and DHT did not alter the E-induced AR mRNA increase. Gilts treated with E plus 10 mg DHT had lesser amounts of immunoreactive ERalpha in cell nuclei of the myometrium and endometrial stroma and a tendency for a decrease in luminal epithelium compared with gilts treated with E. Amounts of immunoreactive ERalpha in glandular epithelium were not influenced by the treatments. Relative amounts of ERalpha and ERbeta mRNAs decreased in the endometrium of gilts treated with E plus 10 mg DHT compared with gilts treated with E. Downregulation of the ERs, particularly ERalpha in the myometrium and endometrial stroma, might be a relevant mechanism in the antagonism of estrogenic effects by DHT in the pig uterus.
    Biology of Reproduction 03/2004; 70(2):297-302. · 4.03 Impact Factor
  • W. F. Pope, H. Cárdenas
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    ABSTRACT: One hundred ewes were monitored twice-a-day for estrus and only those ewes displaying an estrous cycle of 16.5–18.0 days in length were utilized in the experiment. Ewes were subsequently treated with a single injection of 5 or 10mg of prostaglandin F2α (PGF2α) on days 3.0, 3.5, 4.0, 4.5 or 5.0 (n=10 ewes per day) of the estrous cycle (day 0.0=the a.m. or p.m. of first detected estrus) to determine their sensitivity to induction of estrus with prostaglandin F2α. Ewes that were detected in estrus within 24–72h after treatment with PGF2α were considered sensitive to PGF2α treatment. This definition of sensitivity to PGF was inclusive as all ewes either responded within 24–72h after PGF2α treatment or they came into estrus at the expected time based on their previous, untreated, estrous cycle. Results indicated that a significant (P
    Small Ruminant Research - SMALL RUMINANT RES. 01/2004; 55(1):245-248.
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    ABSTRACT: Cytoplasmic maturation refers to a variety of cellular changes that must occur in the oocyte in order to progress through subsequent fertilization and embryonic development. Intracellular concentrations of ATP (ATPi) or glutathione (GSHi), indicative of metabolic activity or the ability of the oocyte to form a male pronucleus and cope with cellular stress, respectively, have been used as markers of cytoplasmic maturation in vitro. In the current study, our objective was to determine if concentrations of ATPi and GSHi in oocytes recovered from three groups of gilts were associated with known differences in developmental competence within these populations. In vivo matured oocytes were surgically recovered 36-38 h after the onset of estrus from first estrous gilts, multi-estrous gilts, and multi-estrous gilts receiving testosterone (1mg/2 ml per day; day 13 to estrus, day 0=day of estrus). Concentrations of ATPi and GSHi were determined using a bioluminescent somatic cell assay kit (luciferin-luciferase reaction) and the dithiobisnitrobenzoic acid (DTNB)-glutathione reductase recycling reaction, respectively. There were no differences (P>0.05) between ATPi concentrations in oocytes from the three groups (1.52 +/- 0.10, 1.51 +/- 0.11, 1.56 +/- 0.11pmol per oocyte). In contrast, oocytes from multi-estrous gilts had higher (P<0.05) concentrations of GSHi (31.53 +/- 1.66 to 33.67 +/- 2.30 pmol per oocyte) than oocytes from first estrous gilts (25.07 +/- 0.82). Administration of testosterone did not affect (P>0.05) GSHi concentrations in oocytes from multi-estrous gilts. Differences in developmental potential between the three groups of gilts were apparently not due to different concentrations of ATPi. However, GSHi concentrations were higher in oocytes from multi-estrous gilts, suggesting that reduced developmental potential of oocytes from first-estrus gilts may be related to insufficient amounts of GSHi. The beneficial effect of exogenous testosterone on the percentage of embryos surviving early gestation does not appear to be due to increased GSHi. Of the numerous potential markers of developmental potential, two were examined in the current study, and GSHi appeared to be useful for assessing porcine oocytes.
    Animal Reproduction Science 10/2003; 78(1-2):123-31. · 1.90 Impact Factor
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    H Cárdenas, W F Pope
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    ABSTRACT: Two experiments were performed to examine the expression of the androgen receptor (AR) gene in the pig uterus. In experiment 1, immunohistochemistry (IHC) was used to determine the distribution of the AR in uterine tIssue of pigs when collected at the first day of estrus (day 0) and the mid-luteal phase (day 12) of the estrous cycle, or early pregnancy (day 12, n=4 gilts per group). In experiment 2, AR immunostaining and AR mRNA in uterine tIssue were compared among ovariectomized gilts (n=4 per group) following treatment for 4 days with daily injections of: (1) progesterone (2 mg/kg bodyweight (BW)), (2) estradiol-17beta (E(2,) 2 micro g/kg BW), (3) E(2) plus progesterone (same dosages as 1 and 2 combined), (4) 5alpha-dihydrotestosterone (DHT, 7 micro g/kg BW), or (5) vehicle (corn oil). Data were analyzed using ANOVA. In experiment 1, nuclear staining for AR in luminal and glandular epithelia was strong and did not differ in intensity between the two locations. Immunostaining of AR in the myometrium was less (P<0.001) intense than in the luminal and glandular epithelia. Nuclei of stromal cells contained AR immunostaining that varied in intensity from strong (mainly in subepithelial stroma) to weak or no staining. Stages of the estrous cycle or early pregnancy did not influence AR immunostaining in the endometrial epithelia and myometrium. In experiment 2, immunostaining of AR in glandular and luminal epithelia and myometrium of ovariectomized gilts treated with vehicle or DHT was less (P<0.05) than in gilts treated with E(2), progesterone, or E(2) plus progesterone. Immunostaining of AR did not differ between ovariectomized gilts treated with vehicle or DHT, or between gilts treated with E(2), progesterone, or E(2) plus progesterone. In both experiments, intensity of AR immunostaining was greater in glandular epithelium located at the adluminal region compared with glandular epithelium located at the basal region of the endometrium. Competitive reverse-transcription PCR (RT-PCR) indicated a stimulatory effect (P<0.01) of E(2) on amounts of AR mRNA in whole endometrium. This increase in AR mRNA after E(2) treatment was not detected when E(2) was combined with progesterone. Endometrial AR mRNA was not influenced by DHT or progesterone relative to vehicle-treated gilts. In conclusion, immunoreactive AR is mainly present in luminal and glandular epithelia of the pig uterus and to a lesser extent in the myometrium, and does not change significantly during the estrous cycle or early pregnancy. Expression of the AR gene in the pig endometrium and myometrium appears to be regulated by E(2) and progesterone.
    Journal of Endocrinology 06/2003; 177(3):461-9. · 4.06 Impact Factor
  • J R Herrick, W F Pope
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    ABSTRACT: Administration of exogenous androgens to pigs during the period of follicular development has been shown to positively affect ovulation rate and embryonic survival. The mechanisms of these actions are not known, but may include direct effects of androgens on the cumulus-oocyte complex (COC). The objective of this experiment was to assess the effects on embryonic development in vitro of exposure of COC to 0.26 and 2.6 microM testosterone (T) or dihydrotestosterone (DHT) during IVM. For IVM, COC were cultured for 44-46 h in protein-free tissue culture medium (TCM) 199 containing 10 IU/ml hCG and eCG and 10 ng/ml EGF. Oocytes were then stripped of cumulus cells, coincubated with 1 x 10(5) sperm/ml in modified TALP for 6 h, and cultured for 8 days in NCSU23. The proportions of oocytes that cleaved (Day 2) or developed to the morula (Day 6) or blastocyst (Day 6-8) stage were not different (P > 0.20) between oocytes exposed to androgens and oocytes not exposed to androgens. These results indicate that exposure to androgens during IVM does not affect the ability of oocytes to cleave or develop up to the blastocyst stage in vitro.
    Theriogenology 11/2002; 58(6):1131-9. · 2.08 Impact Factor
  • H Cárdenas, W F Pope
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    ABSTRACT: Follicle-stimulating hormone (FSH) is an important regulator of follicular development. Some effects of FSH on ovarian follicles might be enhanced by androgens. The main objectives of the present study were to examine expression of the androgen receptor (AR) and FSH receptor (FSHR) in late developing follicles in pigs. Ovaries were collected from gilts on days 13, 15, 17, and 19 of the estrous cycle (day 0 = first day of estrus, n = 4 gilts/day), a period coincident with the follicular phase. One ovary was processed for immunohistochemistry (IHC) of AR. Samples of surface wall from the largest follicles (4-5 per gilt) were dissected from the other ovary, pooled and processed for determination of AR and FSHR mRNAs using reverse transcription-polymerase chain reaction (RT-PCR). Intense AR immunostaining was present in nuclei of granulosa cells of preantral and antral follicles. AR immunoreactivity was also present in the nuclei of oocytes. Weak staining for AR was observed in cells of the theca interna, ovarian surface epithelium, and in most cells of the ovarian stroma. Relative amounts of immunoreactive AR in granulosa cells of late developing follicles, or small antral follicles (< 2 mm), did not differ between days 13, 15, 17, and 19. However, amounts of AR in granulosa cells of small antral follicles was greater (P < 0.05) than in the largest follicles present in the same ovary. The relative amounts of AR mRNA in tissue from the largest follicles on days 13, 15, 17, and 19 did not differ; however, amounts of FSHR mRNA in the same follicles were not different between days 13, 15, and 17, but decreased (P < 0.05) by day 19. Results indicate that during the follicular phase in gilts, the AR protein is mainly present in granulosa cells. Relative amounts of AR protein in granulosa cells and mRNA in walls of late developing follicles did not significantly change from day 13 to 19; however, amounts of FSHR mRNA decreased in preovulatory follicles by day 19 of the estrous cycle.
    Molecular Reproduction and Development 05/2002; 62(1):92-8. · 2.81 Impact Factor
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    ABSTRACT: Treatment with testosterone increases ovulation rate in pigs. The present study was conducted to examine the effects of 5alpha-dihydrotestosterone (DHT), a non-aromatizable androgen receptor ligand, on ovulation rate and amounts of androgen receptor and FSH receptor mRNAs in postpubertal gilts. In Expt 1, ovulation rate in response to daily i.m. injections of 0, 6, 60 or 600 microg DHT kg(-1) body weight from day 13 of the oestrous cycle (day 0 = day 1 of oestrus) to the following oestrus increased with each dose of DHT (P < 0.05). The mean increase in number of corpora lutea ranged from approximately three to 17 over the three dosages of DHT. In Expt 2, gilts treated daily with 60 microg DHT kg(-1) body weight during the early follicular phase (from day 13 to day 16), coincident with follicular recruitment, or the late follicular phase (day 17 to oestrus), had higher (P < 0.05) rates of ovulation compared with gilts that received vehicle, and were not different from gilts treated with DHT from day 13 to oestrus. Percentage recovery of day 3 embryos was not altered when gilts were treated from day 13 to day 16 or from day 17 to oestrus; however, treatment of gilts with DHT from day 13 to oestrus decreased recovery of day 3 (Expt 1) or day 11 (Expt 2) conceptuses. Daily administration of 6 microg DHT kg(-1) body weight to gilts from day 13 of the oestrous cycle to the following oestrus (Expt 3) did not affect the relative amounts of androgen receptor mRNA, but increased (P < 0.05) the amounts of FSH receptor mRNA in preovulatory follicles as determined by RT-PCR. The results of these experiments indicate that androgens may regulate ovulation rate in gilts. One of the roles of androgens might be regulation of the amounts of FSH receptor mRNA in ovarian follicles.
    Reproduction (Cambridge, England) 05/2002; 123(4):527-33. · 3.56 Impact Factor
  • H. Cardenas, W. F. Pope
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    ABSTRACT: Follicular recruitment and atresia are important processes associated with ovulation rate in swine. Follicle-stimulating hormone regulates granu- losa cell division, differentiation, and steroidogenic function, and, as such, significantly influences follicular growth and development. Follicle-stimulating hormone is an inducer of follicular recruitment in swine and an inhibitor of granulosa cell apoptosis, and it seems to be a major regulator of ovulation rate in swine. Although local factors, such as growth factors and steroid hor- mones, might regulate follicular development by con- trolling the expression of gonadotropin receptors or by modulating other related processes, the dominant role of FSH cannot be ignored. Recent results indicate that androgens might be among the local factors regulating
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    ABSTRACT: Objectives were to sequence and examine the expression of the estrogen receptor beta (ERbeta) in the sheep ovary. The sequence of the ovine ERbeta (oERbeta) was determined using reverse-transcription polymerase chain reaction (RT-PCR) and cloning techniques. The reading frame of oERbeta contained 527 amino acids and exhibited high overall homology with cow (98%), rat (88%), and human (88%) ERbeta. In addition, an oERbeta isoform having a 139-base pair deletion (oERbeta1) was identified. The predicted amino acid sequence of this isoform is lacking the ligand-binding and carboxyl-terminal transactivation domains. The oERbeta protein and mRNA were determined in ovaries obtained from ewes on Days 0 (first day of estrus), 2, 6, and 10 of the estrous cycle and Day 30 of gestation. Immunohistochemistry showed that oERbeta protein was located in granulosa cells, the ovarian surface epithelium, endothelium, and Day 2 corpus luteum (CL). Weak immunostaining for ERbeta was detected in the theca interna. Relative steady-state amounts of oERbeta mRNA in the CL were determined using semiquantitative RT-PCR. Amounts of oERbeta mRNA were greater (P < 0.05) during CL formation (Day 2) than at later stages. The oERbeta to oERbeta1 mRNA ratio was lower (P < 0.05) on Day 2 than on Day 10 or Day 30 due to a decrease in amounts of oERbeta1. Results indicate that the oERbeta is a 527-amino acid protein expressed in specific cells of the ovary. Changes in relative amounts of full-length oERB and a deletion isoform in CL occurred during the estrous cycle, suggesting that these two types of ERbeta might regulate estrogen actions during early CL development in sheep.
    Biology of Reproduction 07/2001; 65(1):128-34. · 4.03 Impact Factor
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    ABSTRACT: An effort was undertaken to replace a community of sheep endoparasites that had been classified as resistant to levamisole and albendazole with a community of more susceptible parasites using a dilution approach that could be integrated into the management of a commercial flock. For this study, pastures on this sheep farm were divided into two areas: north and south. Strategically timed anthelmintic treatments combined with pasture management reduced to nondetectable levels the endemic community of anthelmintic resistant parasites in this flock and on these pastures by early summer. A group of 102 ewes, lambs, and rams were experimentally infected with third stage larvae from the more susceptible community of parasites. These sheep then seeded the south pastures with the new parasite community, while sheep on the north pastures maintained the endemic resistant community. Despite its insensitivity as a technique for detecting anthelmintic resistance, fecal egg count reduction tests at the end of the grazing season indicated that the more susceptible parasites were present on the south pastures while resistant parasites were present on the north. The following grazing season, similar protocols were used to introduce the more susceptible parasites onto the north pastures. At the end of the grazing season, fecal egg count reduction tests indicated that the new community of parasites had become established on both groups of pastures of the farm.
    Veterinary Parasitology 07/2001; 97(3):219-25. · 2.38 Impact Factor
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    ABSTRACT: Expression levels of estrogen receptor cofactors (coactivators or corepressors) in specific tissue compartments and cells are thought to influence the expression of estrogen responsive genes and thereby influence overall hormonal responsiveness of target tissues. To date, the presence of cofactors has been reported in tissues from humans, rats and mice. We analyzed the presence and distribution of messenger ribonucleic acids (mRNAs) encoding several transcriptional cofactors in the ovary and uterus of three domestic animal species, the sheep, cow and pig. Northern analysis for cofactors SRC-1, GRIP1, RAC3, p300, RIP140, and SPA showed expression in ovaries from all three species. In addition, lower expression of SRC-1, GRIP1, RAC3, p300, and RIP140 mRNAs was observed during the luteal phase (day 10-12 of the estrous cycle) than at estrus (day 0); however, SPA transcript levels remained unchanged. We then examined expression of mRNAs for changing (SRC-1, RIP140) and constitutively expressed (SPA) cofactors in ovine ovaries. SRC-1 and RIP140 transcripts in corpus luteum were lower compared to the surrounding ovarian tissue. SPA mRNA expression, however, was similar in corpus luteum and surrounding tissues. To determine which ovarian cell types express SRC-1, RIP140, and SPA, in situ hybridization was performed on sheep ovaries. Silver grains corresponding to these cofactors were seen in ovarian granulosa, theca and stromal cells, but appeared to be most abundant in the granulosa cells. Expression of SRC-1 and RIP140 in corpus luteum, however, was reduced compared to expression in follicular cells. Finally, we examined cofactor expression in ovine, bovine, and porcine uterus. Northern blot analysis for SRC-1, GRIP1, RAC3, p300, and RIP140 mRNAs showed higher expression in extracts of the endometrium compared to whole uterus. We provide the first evidence for the presence of estrogen receptor cofactor mRNAs in the ovary and uterus of three domestic animal species. We suggest that coactivators are conserved among species and associated with hormonal responsiveness of reproductive tract tissues in sheep, cow and pig.
    Life Sciences 03/2001; 68(12):1427-38. · 2.56 Impact Factor
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    ABSTRACT: The luteolytic effects of prostaglandin F2 alpha (PGF2 alpha) are thought to be mediated in part by the promotion of an increasingly oxidative cellular environment. Loss of antioxidants is one mechanism by which PGF2 alpha might induce or exacerbate oxidative damage within the corpus luteum. This study was performed to establish whether depletion of vitamin C is an acute effect of PGF2 alpha on the pig corpus luteum and to gain insight into the mechanism of luteal vitamin C loss at luteolysis. Gilts (n = 4) were anaesthetized and both utero-ovarian veins and an ear vein were catheterized. Each corpus luteum on the treated ovary received an intraluteal injection of PGF2 alpha (1 microgram) followed by a sustained release implant containing 100 micrograms of the prostaglandin. The other ovary served as the control and each corpus luteum received corresponding volumes of injection vehicle and blank implant. Blood was collected from the ear vein and both utero-ovarian veins every 15 min beginning 15 min before the onset of treatment. Collection of blood stopped when animals were ovariectomized and corpora lutea were collected at 2 h after treatment. Progesterone and vitamin C (ascorbate) concentrations were measured in tissue and plasma samples. PGF2 alpha-treated luteal tissue had similar progesterone, but significantly lower ascorbate, concentrations when compared with control corpora lutea. PGF2 alpha treatment resulted in a rapid and sustained increase in plasma ascorbate within the treatment-side utero-ovarian vein, while the control utero-ovarian vein and ear vein showed little change in plasma ascorbate during the experimental period. No effect of PGF2 alpha on plasma progesterone was evident. This finding suggests that PGF2 alpha depletes the pig corpus luteum of vitamin C by inducing secretion of the vitamin into the bloodstream. Further studies are necessary to determine whether the depletion of vitamin C that is induced by PGF2 alpha contributes to the demise of the pig corpus luteum.
    J Reprod Fertil 04/1998; 112(2):243-7.
  • Reproduction. 01/1998; 112(2):243-247.
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    ABSTRACT: Three experiments involving 192 crossbred boars evaluated the effects of dietary Se (0 or .5 ppm) and vitamin E (0 or 220 IU/kg) on growth, tissue Se, and alpha-tocopherol concentrations, and on semen quality and its subsequent effect on fertilization rate in mature gilts. Diets formulated used torula yeast and dextrose or cornstarch as the basal feedstuffs and were provided from weaning through sexual maturity. The basal diets averaged .063 ppm Se and 3.46 mg alpha-tocopherol/kg diet. Experiment 1 was a 2 x 2 factorial and conducted as a randomized complete block design in six replicates. Boars were allotted at weaning (initial BW 7.7 kg) with growth and feed performance determined to 145 kg BW. Five boars were killed at weaning and three from each treatment group at periodic intervals to 145 kg BW. Serum and tissue Se and alpha-tocopherol concentration and glutathione peroxidase (GSH-Px) activity were subsequently determined. No performance benefit from either nutrient was demonstrated. Tissue (serum, liver, and testis) GSH-Px activity and Se and alpha-tocopherol concentrations were higher (P < .01) at each period when that respective nutrient fortified the diet. Testis GSH-Px activity increased from weaning to 145 kg BW even when Se was not added to the diet. Experiment 2 was conducted after training three boars from each treatment group of Exp. 1 for semen collection. From 9 mo of age and for a 16-wk period, semen was collected three times weekly and the volume, sperm concentration, motility, and percentage of normal and abnormal sperm were determined. Boars fed either the nonfortified Se or vitamin E diets had sperm with lower motilities (P < .01) and a higher percentage of sperm cells with bent and shoehook tails (P < .01). Diets low in added Se seemed to have a greater detrimental effect on the percentage of motile and abnormal sperm than diets inadequate in vitamin E. Sperm cells had a high concentration of Se and alpha-tocopherol, and a high GSH-Px activity. Experiment 3 was conducted using the boars from Exp. 2; 34 mature gilts were inseminated at 12 and 24 h after estrus. Gilts were killed 5 to 7 d postcoitum and the reproductive tracts were recovered. The semen from boars fed the nonfortified Se diet had a lower fertilization rate of oocytes with fewer accessory sperm penetrating the zona pellucida. The results from these experiments indicate that dietary Se and vitamin E can affect boar semen quality, but the greater effect seemed to be from Se.
    Journal of Animal Science 11/1997; 75(11):2994-3003. · 2.09 Impact Factor