Endocrinology of number of follicular waves per estrous cycle and contralateral or ipsilateral relationship between corpus luteum and preovulatory follicle in heifers

A 3-d extension of the luteal phase occurs in interovulatory intervals (IOIs) with a contralateral relationship between the corpus luteum (CL) and preovulatory follicle with 3 follicular waves (Contra-3W group). Concentrations of FSH, progesterone, LH, and estradiol-17β for the ipsilateral versus contralateral CL and/or follicle relationship and 2 versus 3 waves per IOI were studied in 14 heifers. Follicular waves and FSH surges were designated 1, 2, or 3, according to order of occurrence in the IOI. The day (day 0 = ovulation) of the FSH peak in surge 2 occurred earlier (P < 0.02) in 3-wave IOIs (day 6.3 ± 0.5) than in 2-wave IOIs (day 8.5 ± 0.5). Mean FSH was higher in 3-wave than in 2-wave IOI on 82% of the days in the IOI. Repeatability or individuality in FSH concentration was indicated by a correlation (r = 0.54, P < 0.04) in FSH concentrations between ovulations at the beginning and at the end of the IOI. Concentrations of LH and estradiol increased (P < 0.05) near the beginning of the luteolytic period in 2-wave IOI regardless of the CL and/or follicle relationship. In the Contra-3W group, LH and estradiol remained at basal concentrations concurrently with FSH surge 3 and extension of the luteal phase. The hypotheses were supported that FSH surge 2 occurs earlier in 3-wave IOIs than in 2-wave IOIs and that the development of 3-wave IOIs occurs in individuals with greater FSH concentrations. Extension of the luteal phase in the Contra-3W group was temporally associated with lower concentrations of LH and estradiol.     

Concentrations of ovarian and pituitary hormones following prostaglandin F2 alpha-induced luteal regression in ewes varies with day of the estrous cycle at treatment

Prostaglandin F2 alpha (PGF2 alpha) was injected on d 5, 8 or 11 postestrus in ewes to determine how stage of the estrous cycle would affect PGF2 alpha-induced changes in concentrations of ovarian and pituitary hormones and intervals to the onset of estrus and the preovulatory surge of luteinizing hormone (LH). Initial concentrations of progesterone and average values during the 12 h after PGF2 alpha were related positively to the day of cycle on which PGF2 alpha was administered. Patterns of decline in progesterone after injection of PGF2 alpha were similar among the 3 d. Concentrations of LH in plasma increased in a similar manner from 0 to 12 h in all ewes. After 12 h LH continued to increase, plateaued or declined in ewes treated on d 5, 8 or 11, respectively. Initial concentrations of follicle stimulating hormone (FSH) in plasma were related positively to day of treatment. After treatment with PGF2 alpha, FSH increased within 2 h on d 5 but declined by that time on d 8 or 11. Concentrations of estradiol following treatment did not vary with day. The onset of estrus and the preovulatory surge of LH occurred at 36 and 35, 40 and 45, and 48 and greater than 48 h in ewes treated on d 5, 8 or 11, respectively. It is concluded that: 1) the initial increase in LH is dependent on a decrease in plasma progesterone and 2) differences in patterns of secretion of gonadotropins before the preovulatory surge of LH might be caused by differences in progesterone or progesterone:-estradiol ratio when luteal regression is induced on different days of the estrous cycle.     

Effects of charcoal-extracted, bovine follicular fluid on gonadotropin concentrations, the onset of estrus and luteal function in heifers

A study was conducted to determine the effect of charcoal-extracted, bovine follicular fluid (CFF) on plasma follicle stimulating hormone (FSH) and luteinizing hormone (LH) concentrations, the interval from luteolysis to estrus, and subsequent luteal function in heifers. Fifteen Angus, Simmental and Hereford heifers were allotted by age, weight and breed to a control (C, n = 8) or a CFF (n = 7) group. Heifers received injections of saline or CFF (iv, 8 ml/injection) every 12 h from d 1 (d 0 estrus) through d 5 of the estrous cycle. On d 6, each heifer was injected (im) with 25 mg of prostaglandin F2 alpha (PGF2 alpha). Blood samples were collected every 12 h by venipuncture starting just before the first saline or CFF injection and continuing until estrus. Thereafter, blood samples were collected every other day during the subsequent estrous cycle and assayed for FSH, LH, estradiol-17 beta and progesterone by radioimmunoassay. Injections of CFF had no effect (P greater than .05) on circulating FSH or LH concentrations from d 1 to 5 relative to the C group; however, there was a transient rise (P less than .05) in FSH concentrations 24 h following cessation of CFF injections. This transient rise in FSH was not immediately followed by an increase in plasma estradiol-17 beta concentrations. Although CFF injections did not interfere with PGF2 alpha-induced luteolysis, the interval from PGF2 alpha injection to estrus was delayed (P less than .05) by 5 d in the CFF group compared with the C group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of the dominant follicle aspiration before or after luteinizing hormone surge on the corpus luteum formation in the cow

Luteinizing hormone (LH) surge and follicle rupture act as trigger to start corpus luteum (CL) formation. Thus, we aimed to investigate whether a dominant follicle that has not been exposed to an LH surge can become a functional CL. For this purpose, follicular fluid from the dominant follicles (DF) of cows was aspirated before or after a GnRH-induced LH surge, and subsequent CL formation was observed. Holstein cows were divided into four groups as follows: Luteal phase, a DF was aspirated 7 days after GnRH injection; Pre-LH surge, a DF was aspirated 42 h after PGF(2alpha) injection during the mid luteal phase; Post-LH surge, a DF was aspirated 24 h after GnRH injection following PGF(2alpha); and Intact follicle, ovulation was induced by GnRH injection after PGF(2alpha). Observation of morphological changes in the aspirated follicle using color Doppler ultrasonography and blood sampling was performed on Days 0, 3, 6, and 9 (Day 0 = follicle aspiration). CL formation following DF aspiration was observed only in the Post-LH surge group. In both the Luteal phase and Pre-LH surge groups, however, none of the cows showed local blood flow at the aspirated site or CL formation. Luteal blood flow area, CL volume, and plasma progesterone concentration in the Post-LH surge group were no different from those in the Intact follicle group. The present results clearly demonstrate that rather than follicle rupture, it is the LH surge that is essential for CL formation in cows.     

Effects of ACTH injections during estrus on concentrations and patterns of progesterone, estradiol, LH, and inhibin alpha and time of ovulation in the sow

This study investigated whether injections of ACTH for 48 h, from the onset of the second standing estrus after weaning, had any impact on time of ovulation and patterns of progesterone, estradiol, luteinizing hormone (LH), and inhibin alpha. The studied sows (n=15) were fitted with jugular vein catheters and randomly divided into a control (C group) and an ACTH group. From the onset of standing estrus, the sows were injected (NaCl or synthetic ACTH, 5 microg/kg) every 4h; blood samples were collected immediately before and 45 min after each injection. Ovulation was monitored using ultrasonography. The ACTH-group sows stopped displaying signs of standing estrus sooner after ovulation in their second estrus, but no impact was found on time of ovulation. There were no significant differences in the intervals between LH peak, estradiol peak, and the onset of standing estrus between the C and ACTH groups. The cortisol and progesterone concentrations were significantly elevated (p<0.001) in samples taken 45 min after ACTH injection. There were minor differences in estradiol and LH concentrations between the groups. Overall inhibin alpha concentrations were significantly higher during the treatment period in the ACTH than in the C group, but there were no significant differences between samples taken either 45 min or 4h after injection. In conclusion, injections of synthetic ACTH during estrus in the sow apparently disturb the duration of signs of standing estrus and the hormonal pattern of progesterone, and possibly of inhibin alpha, estradiol and LH.     

黄体期注射PGF2α对母羊血清生殖激素及相关细胞因子的影响

旨在研究黄体期不同阶段注射前列腺激素(PGF2α)对育成母羊生殖激素和生殖相关细胞因子的影响。本研究选择健康、体况良好、体重相近、发情周期正常的湖羊育成母羊60只,用“孕酮栓(MAP)+PMSG”法进行发情周期同步化处理后,选择发情正常的48只母羊随机均分为6组。发情当天记为第0天,黄体前期试验组、中期试验组和末期试验组母羊分别在第6(黄体前期)、11(黄体中期)、16天(黄体末期)注射1 mL PGF2α(0.1 mg),黄体前期对照组、中期对照组和末期对照组母羊分别在第6、11、16天注射1 mL生理盐水,每次注射后0.5、1、2、3 h采血,用于血液指标检测。结果表明,所有试验组和对照组母羊于注射后的0.5~3 h间血清中FSH、LH、PRL、P₄、E₂水平以及TNF-α、IL-1β、IL-6、IFN-β无显著变化(P＞0.05);母羊在黄体期不同阶段注射PGF2α对0.5、1、2、3 h血清中FSH、LH、PRL及IL-1β、IFN-β无显著影响(P＞0.05),前期试验组注射后3 h P₄水平显著低于前期对照组,E₂和IL-6水平显著高于前期对照组(P＜0.05),前期试验组注射后2和3 h TNF-α水平显著高于前期对照组(P＜0.05);中期试验组注射后1 h P₄水平显著低于中期对照组(P＜0.05)。黄体前期注射PGF2α后,前期试验组0.5~ 3 h内FSH、E₂、TNF-α、IL-6整体水平显著高于前期对照组(P＜0.05),P₄整体水平显著低于前期对照组(P＜0.05),对LH、PRL、IL-1β和IFN-β无显著影响(P＞0.05);黄体中期注射PGF2α后,中期试验组0.5~3 h内E₂整体水平显著高于中期对照组(P＜0.05),P₄整体水平显著低于中期对照组(P＜0.05),对FSH、LH、PRL、TNF-α、IL-1β、IL-6和IFN-β无显著影响(P＞0.05);黄体末期注射PGF2α对生殖激素与相关细胞因子没有显著性影响(P＞0.05)。本研究结果表明,PGF2α对黄体的溶解作用存在阶段性差异,母羊在黄体前期对PGF2α的短期应答反应强于黄体中、末期,且黄体前期时,卵巢可以响应PGF2α为卵泡发育营造更佳的发育环境。     

Response of the bovine corpus luteum to increased secretion of luteinizing hormone induced by exogenous gonadotropin releasing hormone

Two experiments were conducted to investigate the response of the bovine corpus luteum to surges of luteinizing hormone (LH) induced by natural gonadotropin-releasing hormone (GnRH) administered twice during the same estrous cycle. In experiment 1, eight mature beef cows, each cow serving as her own control, were injected intravenously (iv) with saline on days 2 and 8 of the cycle (day of estrus = day 0 of the cycle), then with 100 micrograms GnRH on days 2 and 8 of the subsequent cycle. Jugular blood samples were taken immediately prior to an injection and at 15, 30, 45, 60, 120 and 240 min postinjection, to quantitate changes in serum luteinizing hormone. Blood was also collected on alternate days after an injection until day 16 of the cycle, to characterize changes in serum progesterone concentrations. Although exogenous GnRH caused release of LH on days 2 and 8 of the cycle, the quantity of LH released was greater on day 8 (P less than .025). Serum levels of progesterone after treatment with GnRH on day 8 of the cycle did not differ significantly from those observed during the control cycles of the heifers. Because exposure of the bovine corpus luteum to excess LH, induced by GnRH early during the estrous cycle, causes attenuated progesterone secretion during the same cycle, these data suggest that a second surge of endogenous LH may ameliorate the suppressive effect of the initial release of LH on luteal function. Duration of the estrous cycle was not altered by treatment (control, 20.4 +/- .5 vs. treated, 20.4 +/- .4 days).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of estradiol on gonadotrophin release, estrus and ovulation in CIDR-treated beef cattle

The effects of estradiol-17beta (E-17beta) or estradiol benzoate (EB) on gonadotrophin release, estrus and ovulation in beef cattle were evaluated in two experiments. In experiment 1, 16 ovariectomized cows received a previously used CIDR insert from days 0 to 7 and 1mg of EB on day 8; they also received 5mg of E-17beta on days 0 or 1, or 5mg of E-17beta+100mg of progesterone on day 0. There was only an effect of time (P<0.0001) on plasma concentrations of progesterone, estradiol, FSH, and LH. Following treatment with E-17beta, plasma FSH concentrations were suppressed for approximately 36 h, whereas plasma LH concentrations were reduced (P<0.05) for 6 h, but surged within 24 h. Injecting 1mg of EB 24 h after CIDR removal decreased (P<0.02) plasma LH concentrations for 6h, followed by an LH surge at 18 h. In experiment 2, ovary-intact heifers (n=40) received a used CIDR and 5mg of E-17beta+100mg of progesterone on day 0. On day 7, CIDR were removed, PGF given, and heifers received nothing (control) or 1mg of EB 12, 24, or 36 h later. In these groups, plasma LH peaked (mean+/-SEM) 78.0+/-23.0, 37.8+/-8.5, 44.4+/-10.3, and 51.0+/-5.1 h after CIDR removal (means, P<0.001; variances, P<0.001) and intervals from CIDR removal to ovulation were 102.0+/-6.7, 63.6+/-3.6, 81.6+/-3.5, and 78.0+/-4.1h (P<0.05). The interval from CIDR removal to ovulation was shorter and less variable in EB-treated groups; the interval from EB to ovulation was shortest (P<0.05) in the 12-h group. In summary, E-17beta or EB decreased both FSH and LH, but LH increased after 6h (despite elevated progesterone concentrations). Following CIDR removal, 1mg of EB effectively synchronized LH release, and ovulation (in intact cattle), but the interval from CIDR removal to EB treatment affected the time of ovulation.     

Effect of intraluteal injection of endothelin type A receptor antagonist on PGF2alpha-induced luteolysis in the cow

Endothelin-1 (ET-1) is a luteolytic mediator in the bovine corpus luteum (CL), and its action appears to be via endothelin type A receptor (ETR-A). Thus, the aim of the present study was to determine the effect of ETR-A antagonist on PGF2alpha-induced luteolysis in the cow. Cows on days 10-12 of the estrous cycle were subjected to five intraluteal injections of the ETR-A antagonist LU 135252 in saline or only saline at -0.5, 2, 4, 6, and 8 h after PGF2alpha administration (=0 h). Serial luteal biopsies were conducted to determine the expression of mRNA in the luteal tissue. There were no significant differences in the decrease in plasma progesterone (P) concentrations and the mRNA expressions of steroidogenic acute regulatory protein and 3beta-hydroxysteroid dehydrogenase/Delta5, Delta4-isomerase between the ETR-A antagonist-treated group and the control group. However, the start of the decline in CL volume and blood flow area surrounding the CL was delayed for almost two days in the ETR-A antagonist-treated group compared to the control group. The mRNA expression of preproET-1 and endothelin type B receptor increased in both groups, while the ETR-A mRNA remained unchanged. In addition, caspase-3 mRNA expression increased significantly at 24 h in the control group only and its level was higher than that of the ETR-A antagonist-treated group. Thus, the present study suggests that ET-1 regulates structural luteolysis via ETR-A by controlling blood vessel contraction in the CL of the cow.     

A study on ghrelin and LH secretion after short fasting and on ghrelin levels at perioestrual period in dairy cattle

In two experiments, we studied (a) the changes of LH secretion in heifers under different feeding schedules and (b) total ghrelin concentration at oestrus in cows and heifers. In experiment one, synchronized heifers were allocated in three groups (R, regularly fed controls; F, fasted; and F‐F fasted‐fed). One day after the completion of the oestrous induction protocol, group F and F‐F animals stayed without feed for 24 hr; thereafter, feed was provided to R and F‐F cattle; 2 hr later, GnRH was administered to all animals. Blood samples were collected for ghrelin, progesterone, LH and cortisol concentrations. Fasting caused increased ghrelin concentrations in groups F and F‐F, while in response to GnRH, LH surge was significantly attenuated in groups F and F‐F compared to R. In experiment 2, lactating cows and heifers were used. On day 9 of a synchronized cycle, PGF2α was administered, and blood samples were collected twice daily until the third day after oestrus and analysed for progesterone, estradiol, ghrelin, glucose and BHBA concentrations. No difference was recorded between groups in steroids and BHBA concentrations. In comparison to mid‐luteal values, ghrelin concentrations significantly increased at perioestrual period in cows, but not in heifers. This study provides evidence that starving‐induced elevated ghrelin concentrations can have suppressing effect on LH secretion, even after ghrelin's restoration to basal values and that during oestrus, ghrelin secretion is differently regulated in cows and heifers, likely being independent from oestradiol concentrations. Further research is required to identify the determining factors that drive the different regulation of ghrelin secretion in cows and heifers.     

Effect of prostaglandin F2 alpha on the adenylate cyclase and phosphodiesterase activity of ovine corpora lutea

The objective of the present study was to determine the temporal relationships among luteal adenylate cyclase activity, luteal phosphodiesterase activity, luteal progesterone concentration and plasma progesterone concentration during prostaglandin F2 alpha (PGF2 alpha)-induced luteolysis in ewes. Corpora lutea (CL) were removed from cycling ewes on d 9 (d 0 = first day of estrus) at 0, 2, 4, 6, 12 and 24 h (seven to eight ewes/group) after PGF2 alpha administration (im). Jugular blood samples were collected at the time of enucleation of CL and analyzed for progesterone. Plasma and luteal progesterone concentrations were decreased (P less than .05) by 4 and 12 h after PGF2 alpha injection, respectively. Basal adenylate cyclase, luteinizing hormone (LH)-activated adenylate cyclase, guanylylimidodiphosphate [Gpp(NH)p]-activated adenylate cyclase and LH plus Gpp(NH)p-activated adenylate cyclase activities were decreased (P less than .05) by 2 h after PGF2 alpha injection. The decrease in adenylate cyclase activity paralleled the decrease in plasma progesterone concentration over time. Luteinizing hormone stimulated (P less than .05) adenylate cyclase activity relative to basal activity at 0, 2, 12 and 24 h post-PGF2 alpha; whereas, Gpp(NH)p stimulated (P less than .01) adenylate cyclase activity relative to basal activity at each time point. In contrast to the decrease in adenylate cyclase activity, phosphodiesterase activity was increased (P less than .05) at 2 and 4 h post-PGF2 alpha. These results suggest that a decrease in adenylate cyclase activity coupled with an increase in phosphodiesterase activity may decrease the intracellular adenosine 3',5' cyclic monophosphate (cAMP) concentration.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characteristics of developing prolonged dominant follicles in cattle

In cattle, sub-luteal circulating progesterone induces an increase in the frequency of LH pulses, prolonged growth of the dominant follicle, increased peripheral estradiol and reduced fertility. The objective of this study was to examine the earliest stages of development of prolonged dominant follicles, to gain insight into the etiology of this aberrant condition. Heifers were treated with an intravaginal progesterone-releasing device (CIDR) from Day 4-8 post-estrus and PGF2alpha was injected on Day 6 and again 12h later (early prolonged dominant group). Follicular phase (CIDR: Day 4-6, with PGF2alpha) and luteal phase (CIDR: Day 4-8, without PGF2alpha) groups served as controls. As expected, peripheral progesterone in heifers of the early prolonged dominant group was intermediate between luteal and follicular phase groups after luteal regression (P<0.05). On Day 7, the frequency of LH pulses was higher in heifers of the follicular phase and early prolonged dominant groups than the luteal phase group (P<0.05). Dominant follicles (n = 4 per group) were collected by ovariectomy on Day 8 and were similar in size among groups (P>0.05). Estradiol and androstenedione concentrations in the follicular fluid at ovariectomy were higher in the follicular phase and early prolonged dominant groups versus the luteal phase group (P<0.01), whereas progesterone did not differ among groups (P>0.05). Granulosa cells and theca interna isolated from dominant follicles were incubated for 3h with or without gonadotropins or frozen for later analysis of mRNA for steroidogenic enzymes. Luteinizing doses (128 ng/ml) of LH and FSH increased secretion of progesterone (P<0.05) but did not affect secretion of estradiol by granulosa cells in all groups. Low (2 or 4 ng/ml) and luteinizing doses of LH increased secretion of androstenedione by theca interna to a similar extent among groups. Expression of mRNA for P450 side chain cleavage (P450scc), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), P450 aromatase (aromatase) and Steroidogenic Acute Regulatory (StAR) protein by granulosa cells did not differ among groups (P>0.05). Levels of mRNA for P450scc, 3beta-HSD, 17alpha-hydroxylase (17alpha-OH) and StAR protein in theca interna were similar in the follicular phase and early prolonged dominant groups (P>0.05), but lower in the luteal phase group (P<0.05-0.1). In summary, the premature follicular luteinization observed in previous studies after prolonged periods of sub-luteal progesterone was absent in early prolonged dominant follicles, exposed to sub-luteal progesterone for 36 h, and their characteristics resembled those of control follicles during the follicular phase.     

Elevated temperature (heat stress) in vitro reduces androstenedione and estradiol and increases progesterone secretion by follicular cells from bovine dominant follicles

Dairy cattle are susceptible to heat stress-induced reductions in fertility; however, direct effects of hyperthermia on specific reproductive functions are difficult to determine in vivo. The objective of this experiment was to examine the effect of elevated temperature in vitro on follicular steroidogenesis, to gain insight into specific follicular responses associated with heat stress. Dominant follicles were obtained from Holstein heifers on day 6 post-estrus (luteal phase; n = 4) or day 8, 36 h after an injection with 25 mg PGF(2alpha) to induce regression of the corpus luteum (follicular phase; n = 4). Pieces of follicle wall were isolated from dominant follicles and cultured for 96 h with 0, 2 or 100 ng/ml LH or FSH at 37, 39 or 41 degrees C. Concentrations of androstenedione, estradiol and progesterone were determined in culture media collected every 24h. During the last 48 h of culture, basal secretion of androstenedione and estradiol by pieces of follicle wall was lower at 41 degrees C than at 37 or 39 degrees C (P < 0.05). In contrast, cumulative secretion of progesterone by pieces of follicle wall in medium alone was higher at 41 degrees C than at 37 or 39 degrees C (P < 0.05). Pieces of follicle wall responded to treatment with both low (2 ng/ml) and high (100 ng/ml) doses of gonadotropins at all temperatures. However, gonadotropin-induced secretion of androstenedione and estradiol was generally lower, whereas gonadotropin-induced secretion of progesterone was higher at 41 degrees C and sometimes at 39 than at 37 degrees C. The changes in basal steroidogenesis and in responses to gonadotropins suggest that follicular cells begin to luteinize at elevated temperatures in vitro. Premature luteinization of follicular cells in vivo has been associated with reduced fertility in cattle with persistent follicles, suggesting that the premature differentiation of follicular cells observed in the current study may be responsible, in part, for the reduced fertility of dairy cattle under heat-stressed conditions.     

Pituitary Hormone Responses to Luteal Regression after Administration of Prostaglandin F2α-Analogue

As an experiment to elucidate the canine luteal function maintenance system, a PGF2alpha-analogue, fenprostalene (PGF-F), was administered 24 and 54 days after ovulation to induce luteal regression, and the responses of luteinizing hormone (LH) and prolactin (PRL), which are considered to support the canine corpus luteum, were investigated. The plasma progesterone (P(4)) level was rapidly decreased, by which the plasma PRL level was increased, but no change was observed in the plasma LH level. The decrease in the plasma P(4) level after PGF-F administration may have induced positive feedback to the superior system, and stimulated secretion of the pituitary hormones. These findings suggested that the canine corpus luteum is maintained by PRL, not by LH.     

Regulation of pulsatile LH secretion by ovarian steroids in the heifer

Two experiments were conducted to evaluate relationships among luteinizing hormone (LH), estradiol-17 beta (E2) and progesterone secretion during the preovulatory period in the heifer after prostaglandin F2 alpha (PGF2 alpha)-induced regression of the corpus luteum. A second objective was to elucidate the effects of E2 in regulating LH secretion. In Exp. 1, LH, E2 and progesterone concentrations were determined in serial samples collected during the preovulatory period after PGF2 alpha-induced luteal regression in five Red Angus X Hereford heifers. Progesterone declined to 1 ng/ml by 12 h after the second injection of PGF2 alpha. Frequency of LH pulses increased linearly (P less than .01), whereas no change in amplitude of LH pulses was detected before the preovulatory LH surge. This resulted in a linear increase (P less than .01) in mean LH concentrations. Estradiol also increased in a linear manner (P less than .01), and the rise in E2 was parallel to the increase in mean LH concentrations. In Exp. 2, 12 Angus X Hereford heifers were ovariectomized and administered either 13.5- or 27-cm silastic implants containing E2 at ovariectomy. Four heifers served as nonimplanted controls. Thirty-one days after ovariectomy all heifers were bled at 12-min intervals for 6 h. Frequency of LH pulses declined linearly (P less than .03) while mean LH (P less than .09) and pulse amplitude (P less than .01) increased linearly as E2 dose increased. These results indicate that a reduction in progesterone increases the frequency of LH pulses during the follicular phase of the estrous cycle in cattle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of the effectiveness of ovulation synchronization protocol in anestrous and cycling beef cows

Applicability of ovulation synchronization protocol using GnRH and PGF(2alpha) (PGF) injection to anestrous beef cows remains controversial. We compared the effectiveness of the protocol in the anestrous stage of the beef cow with that in the cycling stage using the same animals. Ovaries of five Japanese Black and three Japanese Shorthorn cows were ultrasonographically examined, and blood samples were collected daily for hormonal analyses. Each animal received the protocol twice (Day -6 to -8: GnRH, Day 0: PGF, Day 2: GnRH). Additional blood samples were taken before and after GnRH injection for LH and FSH measurements to evaluate the pituitary function. For the ovarian status at the onset of the protocol cows were divided into anestrous (n=8) and cycling (n=8) stages. There was no significant difference in size of the dominant follicle at the first and second GnRH injections, and in the magnitude of the pituitary response to GnRH between the two stages. However, the size of the corpus luteum and progesterone concentrations at the PGF injection in the anestrous stage were significantly smaller and lower (P<0.01), respectively, and ovulation synchronization rate in the anestrous stage was significantly lower (P<0.05) than in the cycling stage. In conclusion, ovulation synchronization protocol in anestrous beef cows has limited effectiveness.     

Endocrine responses and ovarian changes in inseminated dairy heifers after an injection of a GnRH agonist 11 to 13 days after estrus.

Our objective in this study was to determine endocrine responses and changes in ovarian structures after a single injection of a GnRH agonist in Holstein dairy heifers (n = 38). Heifers were inseminated and received (i.m.) either saline or 200 micrograms of fertirelin acetate once on d 11, 12, or 13 after estrus (d 0). Blood was collected at 15- to 30-min intervals for 6 h after the injection to determine concentrations of LH, FSH, estradiol (E), and progesterone (P) in serum and once daily for 8 to 12 d after the injection to determine concentrations of E and P. Pregnancy rates were 58% (11 of 19) in both treatment groups. Diameter of the corpus luteum and numbers and appearance of ovarian follicles were determined by real-time ultrasonography on d-1 through 5 after injection. No treatment-induced ovulations or changes in the number of ovarian follicles were observed after the injection of the GnRH agonist. More (P less than .05) of the largest follicles within heifers receiving fertirelin acetate showed changes in their appearance on at least the 1st d after injection (6 of 10 vs 1 of 9 control heifers). Fertirelin acetate induced release of LH and FSH from the pituitary within 15 min of injection; both hormones reached peak concentrations at 120 min and then returned to pretreatment concentrations by 300 to 360 min after injection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of feed restriction on reproductive and metabolic hormones in ewes

The goal of this study was to determine the effects of short-term feed withdrawal on reproductive and metabolic hormones during the luteal phase of the estrous cycle in mature ewes. Mature ewes observed in estrus were assigned randomly to control and fasted groups (n = 10 per group Trials 1 and 2). For Trials 1 and 2, control ewes had ad libitum access to feed, whereas fasted ewes were not fed from d 7 through 11 of their estrous cycle; on d 12, all ewes were treated with 10 mg of PGF2alpha, and fasted ewes were gvien ad libitum access to feed. For Trial 1, blood samples were collected daily through fasting and at 2-h intervals following PGF2alpha for 72 h. Serum concentrations of insulin (P < or = 0.002) and IGF-I (P < or = 0.01), but not GH (P > or = 0.60), were decreased during fasting compared with fed ewes. Serum concentrations of 29 (P = 0.02) and 34 kDa (P = 0.04) IGFBP were greater in fasted ewes at 96 h after initiation of fasting than in control ewes. Two control and four fasted ewes in Trial 1 did not exhibit a preovulatory surge release of LH by 72 h. Therefore, Trial 2 was conducted so that the timing of the LH surge could be predicted following the collection of blood samples at 2-h intervals for 112 h and then at 6-h intervals until 178 h following PGF2alpha administration and realimentation. The magnitude of the preovulatory LH surge in Trial 2 was decreased (P = 0.009) and delayed (P = 0.04), and serum concentrations of estradiol were diminished (P < or = 0.03) 12 h before the LH surge in fasted ewes. Ovulation rates were not influenced (P > or = 0.32) by fasting in Trials 1 and 2. Serum concentrations of progesterone in both Trials 1 and 2 were, however, greater (P < 0.001) in fasted than in control ewes. A third trial with ovariectomized ewes was conducted to determine whether the increased serum concentrations of progesterone observed in fasted ewes during Trials 1 and 2 were ovarian-derived. Ovariectomized ewes were implanted with progesterone-containing intravaginal implants and allotted to control (n = 5) or fasted (n = 5) treatment groups and fed as described for Trials 1 and 2. Similar to intact ewes, serum concentrations of progesterone were approximately twofold greater (P < 0.001) in fasted than in control implanted ovariectomized ewes. In summary, feed withdrawal for 5 d during the luteal phase of the estrous cycle increased serum concentrations of progesterone and evoked endocrine changes that could perturb the subsequent estrous cycle.     

Follicular turnover and hormonal association in postpartum goats during early and late lactation

To clarify the effect of lactation period on ovarian follicular activity and associated hormonal levels in goats, six goats were monitored daily by ultrasonographic examination with blood sampling during early (Days 5 to 25; Day 0 was the day of kidding) and late (Days 40 to 60) lactation. While the presence of a corpus luteum of pregnancy retarded follicular growth in the ipsilateral ovary until Days 11-13 postpartum, the total follicular number (TFN) and area (TFA) increased during late lactation due to the significant increase in the number of medium- and large-sized follicles and decrease in the number of small follicles. Four goats showed a similar pattern of follicular development during the period studied characterized by the emergence of five and six waves during the early and late lactation, respectively. The largest follicle diameter of the first three waves monitored during early lactation was significantly smaller as compared with the diameter of those existing during late lactation. TFN showed a positive correlation with FSH but showed a negative correlation with immunoreactive (ir-) inhibin and estradiol during the postpartum period. TFA was positively correlated with ir-inhibin, estradiol and PRL and negatively correlated with FSH during the monitored periods. The plasma levels of ir-inhibin and progesterone were significantly higher during late lactation compared with the levels recorded during early lactation. Ir-inhibin levels showed a significant positive correlation with LH and estradiol during early and late lactation but showed a negative correlation with FSH during the whole lactation period. LH was positively correlated with estradiol and PRL during early and late lactation, respectively. These results suggest that the lactation period has a detrimental effect on ovarian activity during the early postpartum period in goats.     

Changes in bovine luteal progesterone metabolism in response to exogenous prostaglandin F(2alpha)

An experiment was conducted to determine the effect of prostaglandin F2alpha (PGF2alpha) on luteal synthesis of progesterone (P4) and related progestins. Sixteen beef heifers were assigned in equal numbers to four groups in a 2x2 factorial arrangement of treatments. The experiment consisted of two levels of PGF2alpha analog (0 and 500 microg) and two levels of time (4 and 24 h after injection) of corpus luteum collection. All heifers were injected intravenously with saline (2 ml) or PGF2alpha (cloprostenol) on day 8 of the estrous cycle (estrus=day 0). Jugular blood was collected at 0, 1, 2, 3, 4 and 20, 21, 22, 23, and 24 h after injection. Resulting sera were analyzed for P4 by use of radioimmunoassay. Luteal tissue was analyzed by gas chromatography/mass spectrometry for P4, 20beta-hydroxyprogesterone, pregnenolone, and allopregnanolone (3beta-hydroxy-5alpha-pregnan-20-one). Treatment with PGF2alpha reduced serum concentrations of P4 as early as 1 h after injection (P<0.005) and steroid levels remained low over 24 h. Similarly, administration of PGF2alpha caused a decline in luteal P4 (P<0.005), 20beta-hydroxyprogesterone (P<0.10), and pregnenolone (P<0.05). In contrast, treatment with PGF(2alpha) caused an increase in luteal allopregnanolone over time (time x treatment interaction; P<0.05). These data are interpreted to suggest that PGF2alpha promotes conversion of P4 to the metabolite allopregnanolone.