Relationship between uterine secretion of prostaglandin F2 alpha induced by oxytocin and endogenous concentrations of estradiol and progesterone at three stages of the bovine estrous cycle

The purpose of this experiment was to determine whether differences among cows in the ability of oxytocin to stimulate uterine secretion of prostaglandin F2 alpha (PGF2 alpha) were related to the endogenous ovarian steroid environment. Sexually mature heifers were treated with oxytocin (.33 IU/kg BW) at three stages of the estrous cycle: early (d 3 to 5; n = 5), middle (d 10 to 11; n = 5) or late (d 16 to 17; n = 5). To assess uterine responsiveness to oxytocin, concentrations of 13,14-dihydro-15-keto-PGF2 alpha (PGFM) were quantified in jugular venous plasma samples collected at 1/2-h intervals for 8 h postinjection. The ovarian steroid environment at the time of injection was estimated by measuring the concentrations of progesterone and estradiol in jugular venous plasma samples collected at 4-h intervals for 12 h immediately prior to injection. Concentrations of PGFM increased immediately following injection of oxytocin either early or late in the estrous cycle. The response was much less during the middle of the estrous cycle. The magnitudes of response, early and late in the estrous cycle, were similar and greater than that observed during the middle of the estrous cycle (P less than .05). There was a positive relationship (R2 greater than .8; P less than .05) between magnitude of the response to oxytocin and ratio of estradiol to progesterone both early and late in the estrous cycle. Thus, individual differences in uterine secretion of PGF2 alpha in response to oxytocin were related to stage of the cycle and to differences in the endogenous ovarian steroid environment within each stage of the estrous cycle.     

Microvascularization and angiogenic activity of equine corpora lutea throughout the estrous cycle

Corpus luteum growth and endocrine function are closely dependent on the formation of new capillaries. The objectives of this study were to evaluate (i) tissue growth and microvascular development in the equine cyclic luteal structures; (ii) in vitro angiogenic activity of luteal tissues in response to luteotrophic (LH, PGE₂) and luteolytic (PGF₂) hormones and (iii) to relate data to luteal endocrinological function. Our results show that microvascular density was increased in the early and mid luteal phase, followed by a fall in the late luteal phase and a further decrease in the corpus albicans. Hyperplasia of luteal tissue increased until the mid luteal phase and it was followed by tissue regression. Luteal explants were cultured with no hormone added, or with PGF₂, LH, PGE₂, LH + PGE₂ or LH + PGF₂. Media conditioned by equine luteal tissue from different stages of the luteal phase were able to stimulate mitogenesis of bovine aortic endothelial cells (BAEC), suggesting the presence of angiogenic activity. No difference was observed among luteal structures on their mitogenic capacity, for any treatment used. Nevertheless, Late-CL conditioned-media with PGF₂ showed a significant decrease in BAEC proliferation (p < 0.05) and LH + PGF₂ a tendency to reduce mitogenesis. Thus, prostaglandin F₂ may play a role on vascular regression of the CL during the late luteal phase in the mare. These data suggest that luteal angiogenesis and vascular regression in the mare are coordinated with the development of non-vascular tissue and might be regulated by many different factors.     

VEGF system expression in different stages of estrous cycle in the corpus luteum of non-treated and superovulated water buffalo

Water buffaloes are easily adaptable animals, whose raising and economical exploitation have been growing in the last three decades all over the world. Hyperstimulation of ovarian function in this species is a common technique aiming to improve reproductive performance. Superovulatory treatment affects corpus luteum (CL) function, which is highly correlated to angiogenic process. The aim of this study was therefore to assess the temporal protein and mRNA expression of VEGF and its receptors in the CL of non-treated and superovulated buffaloes. For that purpose blood samples and CL from 36 healthy (30 untreated, groups 1–5, and 6 superovulated, group 6) non-pregnant buffaloes were collected and the samples were divided into 6 groups according to the age of CL. Plasma samples were submitted to RIA to measure progesterone concentration and CL were subjected to immunohistochemistry and real time PCR for VEGF (vascular endothelial growth factor), Flt-1 (fms-like tyrosine kinase receptor 1) and KDR (kinase insert domain containing region). The VEGF system protein and mRNA expression during CL life span of untreated animals showed a specific time-dependent profile, although protein did not always reflect mRNA concentrations. VEGF expression in luteal cells was high correlated to plasma progesterone levels. Superovulated CL showed a significant increase of the VEGF-system protein and a significant decrease of mRNA expression compared to untreated animals in the same stage of the oestrous cycle. We conclude that VEGF, Flt-1 and KDR protein and mRNA expression in buffalo CL is dependent of estrous cycle stage and superovulatory treatment is able to increase the translation rate of this system.     

Biological activity of LH during the peripartum period and the estrous cycle of the ewe

Biological activity of luteinizing hormone (LH) is related to the degree of glycosylation of the glycoprotein hormone. The objectives of this study were to determine changes in biologically (BLH) and immunologically (ILH) active LH concentrations in plasma (in vitro bioassay and radioimmunoassay, respectively) and in the ratio of BLH to ILH (B:I) during the peripartum period and during the estrous cycle of the ewe. Blood samples were collected daily 4 days before through 4 days after parturition and during one estrous cycle. Also, samples were collected at 15-min intervals for 6 hr on Days 3 and 12 of the estrous cycle to quantify the influence of an elevated plasma concentration of progesterone (P) on the episodic secretion profiles of BLH. Progesterone concentration was determined on the 4th days pre- and post-partum, on each day of the estrous cycle and at hourly intervals on Days 3 and 12 of the cycle to investigate the hypothesis of an inverse relationship between P and BLH. The BLH and ILH concentrations were low during the peripartum period, and the B:I ratio did not increase by the 4th day postpartum. Mean ILH concentration was greater (P less than .05) in the postpartum than during the prepartum period. During the estrous cycle, mean daily B:I ratio was consistently above unity except for the day of estrus. The pre-ovulatory LH surge (BLH and ILH) was associated with a decrease (P less than .05) in the mean B:I ratio to 0.0065. Mean concentrations of BLH and ILH in plasma samples collected every 15 min on Day 12 were similar to Day 3 of the cycle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Synchronization of estrus in dairy goats given norgestomet and estradiol valerate at various stages of the estrous cycle.

Dairy goats were given subcutaneous implants with 3 mg of norgestomet (NOR) and IM injections of 0.625 mg of estradiol valerate and 0.375 mg of norgestomet on day 0 of the estrous cycle (estrus; NOR 0, n = 18), on postestrus day 4 (NOR 4, n = 18), or on postestrus day 11 (NOR 11, n = 15). Ear implants were removed after 9 days. Mean (+/- SE) hours from removal of ear implants to onset of estrus and proportion of goats responding were 36 +/- 3.8 and 83%, 33 +/- 4.0 and 61%, and 36 +/- 2.7 and 93% for groups NOR 0, NOR 4, and NOR 11, respectively. There were no significant differences between treatment groups in time to onset of estrus. The percentage of goats in group NOR 11 that had signs of estrus was significantly greater than the percentage of goats in group NOR 4. Of the goats in groups NOR 0, NOR 4, and NOR 11 that had signs of estrus, 53, 55, and 86%, respectively, had onset of behavioral estrus between 24 and 48 hours after implant removal. All goats that had signs of estrus had onset of behavioral estrus between 12 and 72 hours after implant removal. Mean (+/- SE) hours from removal of ear implants to time of peak concentrations of luteinizing hormone (LH) were 49 +/- 4.1, 49 +/- 3.8, and 49 +/- 4.0 for groups NOR 0, NOR 4, NOR 11, respectively (not different). The percentage of goats in group NOR 11 that had LH peaks was significantly greater than the percentage of goats in group NOR 4.(ABSTRACT TRUNCATED AT 250 WORDS)     

Synchronization of estrus in dairy goats treated with prostaglandin F at various stages of the estrous cycle.

Dairy goats were given IM injections of 125 micrograms of cloprostenol sodium on day 6 of the estrous cycle (prostaglandin F [PGF] 6, n = 22) or day 12 of the estrous cycle (PGF 12, n = 26). Mean +/- SE hours from injection to onset of behavioral estrus and proportion of goats responding were 46 +/- 4.2 (range, 12 to 88 hours) and 95% and 48 +/- 2.9 (range, 34 to 68 hours) and 100% for groups PGF 6 and PGF 12, respectively. There was no significant difference between the groups in mean time to onset of estrus, but variances about the means were different. Of the does in groups PGF 6 and PGF 12, 67 and 85%, respectively, had signs of onset of estrus between 36 and 60 hours after administration of PGF. Mean (+/- SE) hours from injection to time of peak concentrations of luteinizing hormone (LH) were 62 +/- 3.1 and 64 +/- 2.1 for groups PGF 6 and PGF 12, respectively. Of the does in groups PGF 6 and PGF 12, 86 and 100%, respectively, had LH peaks. Of the does in groups PGF 6 and PGF 12, 68 and 77%, respectively, had peak concentrations of LH between 48 and 72 hours after administration of PGF. All does in groups PGF 6 and PGF 12 had concentrations of progesterone greater than or equal to 1.0 ng/ml on the day of administration of PGF. Concentrations decreased to less than 1.0 ng/ml by 48 hours after injection in all does except 1 in group PGF 6.(ABSTRACT TRUNCATED AT 250 WORDS)     

Control of the estrous cycle in the mare

All current approaches to manipulating the reproductive biology of the nonpregnant mare are discussed.     

Relationship between endogenous estradiol-17 beta and estrous behavior in heifers

Thirty-five Holstein heifers were used to examine the relationship between endogenous estradiol-17 beta and estrous traits. During a non-superovulation period (NSP), estrous cycles were synchronized and during the periovulatory stage blood samples were collected every 6 h for 120 h for subsequent determination of estradiol-17 beta and progesterone. In addition, continuous observation for estrous behavior was performed for 98 h. A gonadotropin-induced superovulation period (SP) was begun 12 d after estrus was detected during NSP. Heifers were injected with FSH twice daily for 4 d and single injections of prostaglandin were given on d 14 and 15. Beginning at d 14, blood samples were collected every 6 h for 120 h for subsequent determination of estradiol-17 beta and progesterone. Continuous observation for estrous behavior was performed for 98 h. Peak estradiol-17 beta was greater during SP than during NSP (49.0 +/- 3.1 vs 12.9 +/- 3.0 pg/ml serum). Thirty-three and 31 of the 35 heifers were in estrus during NSP and SP, respectively; duration of estrus was 2.3 h longer during SP than during NSP. However, number of behavioral interactions during estrus did not differ between NSP and SP. In conclusion, estrous traits were similar, whereas peak estradiol-17 beta concentrations were markedly different between NSP and SP.     

Ovarian follicular populations at two stages of an estrous cycle in heifers given high energy diets

Sixteen crossbred heifers were assigned at random on d 3 of an estrous cycle to continue to receive the medium quality hay (control diet) or to be flushed with a grain ration (high-energy diet) until slaughtered on d 12 or 16. The ovarian follicles were counted and measured using routine histological techniques. Both the nonatretic and atretic (more than four pyknotic bodies) antral follicles (greater than .14 mm) were separated into six classes according to diameter size. The mean number of nonatretic follicles measuring .29 to .67 mm and .68 to 1.57 mm in heifers on a high-energy diet increased from 30.8 and 8.1 on d 12 to 57.5 and 15.5 on d 16, whereas in heifers on a control diet it decreased from 43.1 and 13.0 to 28.1 and 6.5, respectively (energy in diet X day of estrous cycle interaction, P less than .05). The mean number of nonatretic follicles measuring 3.68 to 8.56 mm decreased from 2.1 to 0 between d 12 and 16 in heifers on a high-energy diet while it remained the same at 0 and .5, respectively, for heifers on a control diet (energy in diet X day of cycle interaction, P less than .05). Of the total number of follicles (mean = 161) 38.2% were atretic. Mean number of atretic follicles measuring 1.58 to 3.67 mm in heifers on a high-energy diet increased from 14.8 at d 12 to 31.0 at d 16 whereas in heifers on a control diet it decreased from 27.4 to 13.3, respectively (energy in diet X day of cycle interaction, P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between the limbic system and gonadal function. Quantifiable morphokinesis of the medial amygdaloid nucleus with reference to the estrous cycle]

Karyovolumetric investigation showed that in rat the neurons of the limbic system which are associated with the Pars caudalis of the nucleus amygdaloideus medialis followed a morphokineses which was quantifiable in relation to the oestral cycle. The cell nuclei differed significantly in volume between cycle phases, which was interpreted as morphological correlation between variations in neuronal activity of metabolism. It was thought to be another point in support of the relationship between the amygdala and central-nervous structures of the cerebro-pituitary-gonadal axis.     

Mechanical and morphometric analysis of the third carpal bone of Thoroughbreds

The third carpal bone (C3) was collected from both forelimbs of 27 Thoroughbreds. On the basis of age, training, and history, specimens were assigned to 1 of 5 groups: yearling, untrained horses (group 1, n = 4); 2- to 3-year-old, untrained horses (group 2, n = 7); trained 2-year-old horses (group 3, n = 6); trained 3-year-old horses (group 4, n = 6); and 3-year-old, trained horses with carpal pathologic features (group 5, n = 4). A transverse section of subchondral bone 5-mm thick was cut in a precise fashion 10 mm below the proximal articular surface of all specimens. After high-detail radiography was done, indentation testing was performed on the proximal surface of the section at points 5 mm apart. The stiffness of the subchondral cancellous bone was determined from the slope of the load vs displacement curve. Topographic plots of stiffness measurements were compared with radiographs of each specimen. Point determinations were averaged to derive measures for the radial and intermediate facets, and for regions 5, 10, 15, and 20 mm from the dorsal margin of C3. Area fraction (1-p; p = porosity) was measured for the radial and intermediate facets, using an automated image analysis system. Significant (P less than 0.05) increases in stiffness and area fraction were found in the C3 from trained horses (groups 3 to 5), compared with untrained horses (groups 1 to 2). Stiffness and area fraction of the radial facet of pathologic C3 were significantly higher than the same variables measured in C3 from any other group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Biological activity of Luteinizing Hormone (LH) during the estrous cycle of mares

Equine luteinizing hormone (LH) contains a high proportion of carbohydrate (27%) and large compliment of sialic acid residues (6%), and anterior pituitary stores exhibit high physiochemical polymorphism. We initiated studies to determine whether polymorphism in the pituitary was reflected in secretory forms of LH as divergence between biopotency and immunoreactivity. Serum samples were collected daily from mares throughout the estrous cycle. In addition, the dynamics of LH secretion was monitored during intensive collection periods during estrus and the mid-luteal phase. Immunoreactive LH (iLH) was evaluated using conventional radioimmunoassay procedures. The biological activity of LH (bLH) was determined using a mouse interstitial cell-testosterone bioassay. The relative biopotency of equine LH (bLH/iLH,B/I) is significantly (P<0.05) greater during the preovulatory (1.69 ± 0.11) and luteal (1.45 ± 0.07) phases of the cycle than during the period of luteal regression (1.11 ± 0.07). These data indicate that the secretory forms of equine LH vary through the estrous cycle and suggest that gonadal steroids may influence the processing of the hormone at the pituitary or at an extra-hypophyseal locus.     

Relationship between estrous behavior in pregnant mares and the presence of a female conceptus

Unsolicited reports of estrous behavior in mares thought to be pregnant were received from owners or caretakers of Arabian mares. Estrous behavior was confirmed and mares were examined for pregnancy. Gender of the conceptus was determined at foaling in 11 mares in which estrous behavior was confirmed while an apparently viable, ultrasonically normal-appearing conceptus was present. In 9 mares in which the day of ovulation was known (Day 0), the estrous behavior occurred on Day 12, 13 or 14 (5 mares), Day 18 or 20 (2 mares), Day 40 (1 mare) and Day 60 (1 mare). In another study, 55 pony mares were observed for estrous behavior every 3 days for 20 minutes during Days 11 to 40. Estrous behavior was observed in 1 mare (2%) on Day 24. Combined for the 2 studies, the incidence of a female conceptus (12/12) was greater (P<0.01) than the incidence of a male conceptus (0/12) in mares that exhibited estrous behavior.     

Follicle stimulating hormone pattern and luteal function in ewes receiving bovine follicular fluid during three stages of the estrous cycle

The objectives of this study were to determine 1) the ability of charcoal-extracted bovine follicular fluid (bFF) to suppress endogenous follicle stimulating hormone (FSH) at various stages of the estrous cycle and 2) the effects of suppression of FSH on luteal function and lengths of the current and subsequent estrous cycles. Twenty-six mature ewes were assigned randomly to receive 5 ml of either bFF or saline, subcutaneously, at 8-h intervals on d 1 through 5 (bFF n = 6; saline n = 3), d 6 through 10 (bFF n = 6; saline n = 3) or d 11 through 15 (bFF n = 6; saline n = 2) of the estrous cycle (d 0 = estrus). Blood was collected daily beginning at estrus and continued until the third estrus (two estrous cycles) or 40 d; more frequent samples were collected 2 h prior to initiation of treatment (0600), hourly for the first 8 h of treatment, then every 4 h until 0800 on the first day after treatment, and finally at 1600 and 2400 on that day. Plasma concentrations of FSH were lower (P less than .001) in bFF-treated than in saline-treated ewes. Treatment with bFF reduced (P less than .05) plasma concentrations of progesterone during the current but not during the subsequent estrous cycle. Treatment with bFF did not affect plasma concentrations of estradiol-17 beta. Administration of bFF on d 11 through 15 of the estrous cycle lengthened the interval from the decline in progesterone to estrus and the inter-estrous interval by approximately 3 and 4 d, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Luteinizing hormone-releasing hormone fusion protein vaccines block estrous cycle activity in beef heifers

Two LHRH fusion proteins, thioredoxin and ovalbumin, each containing seven LHRH inserts were tested for their ability to inhibit estrous cycle activity. The objective was to evaluate immune and biological responses from alternating the two fusion proteins in an immunization schedule. One hundred ten heifers were divided equally into 11 groups. Two control groups consisted of either spayed or intact, untreated heifers. Heifers in the other nine groups were immunized on wk 0, 4, and 9. Treatments were immunizations of the same protein throughout or alternating the proteins in different booster sequences. Blood was collected weekly for 22 wk, and serum was assayed for concentrations of progesterone and titers of anti-LHRH. At slaughter, reproductive tracts were removed from each heifer and weighed. Heifers with >or=1 ng/mL of progesterone were considered to have a functional corpus luteum and thus to have estrous cycle activity. All LHRH-immunized groups of heifers had a smaller (P < 0.05) proportion of heifers showing estrous cycle activity after 6 wk than the intact, untreated control group. There was no difference in number of heifers cycling between the immunized groups and the spayed heifers during wk 9 to 22. Anti-LHRH did not differ among immunized groups during wk 1 to 9. Starting at wk 10 and continuing through the conclusion of the study, there was an overall difference among treatment groups for anti-LHRH (P < 0.05). Uterine weights differed among treatments (P < 0.05), with intact control animals having heavier uteri than all other groups (P < 0.05). Uterine weights were negatively correlated with maximum LHRH antibody binding (r = -0.44). In summary, the LHRH fusion proteins were as effective as surgical spaying in suppression of estrous cycle activity, but alternating the two proteins in an immunization schedule did not enhance the immunological or biological effectiveness of the vaccine.