Pregnancy rates to fixed-time AI in Bos indicus-influenced beef cows using PGF2α with (Bee Synch I) or without (Bee Synch II) GnRH at the onset of the 5-day CO-Synch + CIDR protocol.

Objectives were to 1) characterize fixed-time AI (FTAI) pregnancy rates using the 5-Day CO-Synch + CIDR protocol in mature, suckled Bos indicus-influenced beef cows, 2) compare FTAI pregnancy rates in the latter to a modified version (5-Day Bee Synch + CIDR; Bee Synch I) that included treatment with prostaglandin F2α (PGF2α) at CIDR insertion on Day 0, and 3) test the hypothesis that elimination of both GnRH-1 at the onset of synchronization and the double dose of PGF on Day 5 (Bee Synch II) would not reduce FTAI pregnancy rates compared to Bee Synch I. For Experiment 1-trial 1, Brahman x Hereford (F-1) cows (n = 168) at least 40 d postpartum (PP; r = 40-92 d) at the time of CIDR insertion were administered the 5-Day CO-Synch + CIDR protocol with FTAI at 72 h after CIDR removal. Pregnancy rates to FTAI averaged 34.9 ± 1.9%. In Experiment 1-trial 2, fall- and spring-breeding Brahman x Hereford (F-1) beef cows (n = 269) were stratified by days PP and assigned randomly to receive either the 5-Day CO-Synch + CIDR (n = 136) or Bee Synch I (n = 133) protocol, with FTAI at 66 h after CIDR removal. Pregnancy rate to FTAI was greater (P < 0.05) in Bee Synch I (52.6 ± 0.9%) than in the 5-Day CO-Synch + CIDR procedure (40.4 ± 5.7%). For Experiment 2, 422 mature Braford, Brangus, Nelore x Brahman, and Brahman crossbred cows (Bos indicus proportion unknown) at 4 locations were treated with Bee Synch I, with FTAI at 66 h. Overall FTAI pregnancy rate averaged 51.7 ±â€¯2.1%. Finally, from 2013 through spring 2018, we used a switchback design using fall- and spring-breeding herds to compare Bee Synch I (402 observations) to Bee Synch II (393 observations). Overall frequency of detected estrus at 66 h using ESTROTECT™ breeding indicator patches was 57.2 ±â€¯2.4%, conception rates of those detected in estrus was 64.4 ±â€¯3.5%, and FTAI pregnancy rates averaged 52.3 ±â€¯2.4%, none of which differed between treatments. Moreover, pregnancy rates to FTAI in both treatments did not differ in cows synchronized between 40 and 80 d PP but increased after 80 d PP (P < 0.05). Bee Synch II, which eliminates GnRH-1 and the double dose of PGF2α on Day 5, results in FTAI pregnancy rates essentially identical to Bee Synch I but reduces synchronization costs and avoids the need for off-label (double dose PGF2α) drug use.

Ovarian follicular and luteal characteristics in Bos indicus-influenced beef cows using prostaglandin F2α with or without GnRH at the onset of the 5-day CO-Synch + controlled internal drug release (CIDR) protocol.

A modification of the standard 5-day CO-Synch + CIDR procedure (5-day Bee Synch + CIDR; Bee Synch), developed for use in Bos indicus-influenced cows, utilizes the addition of prostaglandin F2α (PGF) on Day 0 of the protocol to eliminate mature corpora lutea (CL) and fixed-time AI (FTAI) at 66 h. Objectives were to test the hypothesis that elimination of GnRH on Day 0 (GnRH-1) does not impact significantly the synchronized development of a dominant follicle for presumptive FTAI. Seventy-one estrous cycling Brangus and Brahman x Hereford suckled cows were used in two replicates (35-36/replicate). Following stratification, cows were assigned randomly to a 2 × 3 factorial arrangement of treatments involving two truncated (no FTAI or GnRH-2) versions of Bee Synch (Bee Synch It and IIt), each begun 3, 7, and 10 days post-ovulation. Cows in Bee Synch It received 100 µg GnRH (GnRH-1), 25 mg PGF, and a CIDR on Day 0, whereas cows assigned to Bee Synch IIt received the same treatment but without GnRH-1. All cows received 50 mg PGF on Day 5 at CIDR removal. Synchronized new follicular wave emergence (NFWE; days 1-4) was observed in 68.6 and 38.9% of Bee Synch It and IIt, respectively (P = 0.01). This increased to 93.3% and 72.2%, respectively, if days 0-4 were considered. Inclusion of GnRH at CIDR insertion improved synchronized NFWE but size of the largest follicle at 66 h, the normal time of FTAI, did not differ due to treatment or day of the estrous cycle.

Effects of acute fasting and age on leptin and peroxisome proliferator-activated receptor gamma production relative to fat depot in immature and mature pigs.

Leptin and peroxisome proliferator-activated receptor gamma (PPARγ) are adipogenic proteins that are actively involved in metabolic homeostasis of fat. Recently, it was reported that fat tissue in humans and rodents differs in metabolic activity relative to anatomical location of the fat tissue (i.e. depots) and animal age. Hence, we hypothesized that leptin and PPARγ production in various fat depots in female pigs differs in response to acute fasting, and that these responses vary with physiological maturity of the animal. Sixteen intact crossbred immature female pigs [prepubertal (PP); 132.2 ± 4.1 days] and 16 sexually mature female pigs (M; 224 ± 7.4 days) housed in an open-air, concrete slab, sheltered barn were randomly assigned to either Control or Fasted treatments. Control pigs (PP, n = 8; M, n = 8) had ad libitum access to feed, while Fasted pigs (PP, n = 8; M, n = 8) were denied access to feed from the onset of the study (0 h) to euthanasia at 72 h. Immediately post-mortem, fat samples were collected from the subcutaneous, pelvic, kidney, and heart (M pigs only) fat depots and analysed for leptin and PPARγ mRNA and protein content. Acute fasting decreased mean leptin mRNA tissue content in a depot specific manner in M pigs (p < 0.01), while mean leptin protein concentrations in fat tissues did not differ with fat depot or age of the pig. Furthermore, acute fasting did not affect mean PPARγ mRNA tissue content in a fat depot or age dependent manner. Mean concentrations of PPARγ protein in fat depots tended to be greater in M vs. PP pigs (p = 0.07). We suggest that these data provide evidence that acute fasting has a greater effect on leptin than PPARγ production in a fat depot dependent manner in M pigs, which may be indicative of changing physiological demands as an animal matures.

Ovarian, hormonal, and reproductive events associated with synchronization of ovulation and timed appointment breeding of Bos indicus-influenced cattle using intravaginal progesterone, gonadotropin-releasing hormone, and prostaglandin F2alpha.

The objectives of this study were to 1) compare cumulative pregnancy rates in a traditional management (TM) scheme with those using a synchronization of ovulation protocol (CO-Synch + CIDR) for timed AI (TAI) in Bos indicus-influenced cattle; 2) evaluate ovarian and hormonal events associated with CO-Synch + CIDR and CO-Synch without CIDR; and 3) determine estrual and ovulatory distributions in cattle synchronized with Select-Synch + CIDR. The CO-Synch + CIDR regimen included insertion of a controlled internal drug-releasing device (CIDR) and an injection of GnRH (GnRH-1) on d 0, removal of the CIDR and injection of PGF2alpha (PGF) on d 7, and injection of GnRH (GnRH-2) and TAI 48 h later. For Exp. 1, predominantly Brahman x Hereford (F1) and Brangus females (n = 335) were stratified by BCS, parity, and day postpartum (parous females) before random assignment to CO-Synch + CIDR or TM. To maximize the number of observations related to TAI conception rate (n = 266), an additional 96 females in which TM controls were not available for comparison also received CO-Synch + CIDR. Conception rates to TAI averaged 39 +/- 3% and were not affected by location, year, parity, AI sire, or AI technician. Cumulative pregnancy rates were greater (P < 0.05) at 30 and 60 d of the breeding season in CO-Synch + CIDR (74.1 and 95.9%) compared with TM (61.8 and 89.7%). In Exp. 2, postpartum Brahman x Hereford (F1) cows (n = 100) were stratified as in Exp. 1 and divided into 4 replicates of 25. Within each replicate, approximately one-half (12 to 13) received CO-Synch + CIDR, and the other half received CO-Synch only (no CIDR). No differences were observed between treatments, and the data were pooled. Percentages of cows ovulating to GnRH-1, developing a synchronized follicular wave, exhibiting luteal regression to PGF, and ovulating to GnRH-2 were 40 +/- 5, 60 +/- 5, 93 +/- 2, and 72 +/- 4%, respectively. In Exp. 3, primiparous Brahman x Hereford, (F1) heifers (n = 32) and pluriparous cows (n = 18) received the Select Synch + CIDR synchronization regimen (no GnRH-2 or TAI). Mean intervals from CIDR removal to estrus and ovulation, and from estrus to ovulation were 70 +/- 2.9, 99 +/- 2.8, and 29 +/- 2.2 h, respectively. These results indicate that the relatively low TAI conception rate observed with CO-Synch + CIDR in these studies was attributable primarily to failure of 40% of the cattle to develop a synchronized follicular wave after GnRH-1 and also to inappropriate timing of TAI/GnRH-2.

Evidence that lamprey GnRH-III does not release FSH selectively in cattle.

Experiments were performed to test the hypothesis that lamprey GnRH-III (lGnRH-III) selectively releases FSH. Primary cultures of bovine adenohypophyseal cells were treated with mammalian GnRH (mGnRH) and lGnRH-III (10(-9), 10(-8), 10(-7) and 10(-6) M) or control media in Experiment 1. All doses of mGnRH and the two highest doses of lGnRH-III stimulated (P < 0.001) a non-selective release of LH and FSH. In Experiments 2-4, Latin Square designs were utilized in vivo to examine whether physiological and hormonal milieu regulate putative selective effects of lGnRH-III. In Experiments 2 and 3, ovariectomized cows with basal levels of estradiol only (Experiment 2) or in combination with luteal phase levels of progester-one (Experiment 3) were injected with mGnRH and lGnRH-III (0.055, 0.11, 0.165 and 1.1 microg/kg body weight (BW) and saline. All doses of mGnRH released (P < 0.001) LH and FSH, but only the highest dose of lGnRH-III stimulated (P < 0.001) a non-selective release of both LH and FSH (Experiment 3). For Experiments 4A and 4B, intact, mid-luteal phase cows were injected with mGnRH and lGnRH-III (1.1 microg/kg BW; Experiment 4A), lGnRH-III (1.1 and 4.4 microg/kg BW; Experiment 4B) and saline. As before, mGnRH released (P < 0.001) both LH and FSH at all doses. In contrast, lGnRH-III at the highest dose released (P < 0.001) LH but not FSH. These findings suggest that lGnRH-III may act as a weak competitor for the mGnRH receptor and do not support the hypothesis that it selectively releases FSH in cattle.

Leptin acts at the bovine adenohypophysis to enhance basal and gonadotropin-releasing hormone-mediated release of luteinizing hormone: differential effects are dependent upon nutritional history.

Recombinant ovine leptin (oleptin) stimulates an acute increase in the secretion of LH in fasted, but not in normal-fed, cows through an augmentation of the magnitude of individual pulses of LH. Herein, we tested the hypothesis that this effect could be accounted for by functional changes at the adenohypophyseal (AP) level. Eleven ovariectomized, estradiol-implanted cows were assigned to one of two dietary groups: normal-fed (n = 6) and fasted (fasted for 72 h; n = 5). After the animals were killed, the adenohypophyses were collected and AP explants were perifused with Krebs-Ringer bicarbonate buffer (KRB) for a total of 6.5 h, including a 2-h treatment at 2.5 h with KRB or increasing doses of oleptin and a challenge at 4.5 h with 50 ng of GnRH. To test for effects of leptin at the hypothalamic level, explants encompassing the medial basal hypothalamus-infundibular complex (HYP) were incubated in KRB alone (control) or in KRB containing 1000 ng of oleptin. Basal release of LH from AP explants treated with leptin was greater (P < 0.02) than that from control-treated explants in fasted, but not in normal-fed, cows. To the contrary, leptin-treated explants from normal-fed, but not from fasted, cows released more (P < 0.001) LH in response to GnRH than control-treated tissues. Neither fasting nor leptin affected (P > 0.1) the secretion of GnRH from HYP explants. These observations support the hypothesis that leptin modulates the secretion of LH in mature cows, to a large extent, by its direct actions at the AP. Differential manifestations of these effects are dependent upon nutritional history.

Serum leptin and its adipose gene expression during pubertal development, the estrous cycle, and different seasons in cattle.

Circulating concentrations of leptin and IGF-I, leptin gene expression, and serum binding of [126I]ovine leptin in cattle during pubertal development, as well as leptin gene expression and circulating concentrations of leptin during the estrous cycle and different calendar seasons, were investigated. Multivariate regression analysis was utilized to evaluate temporal changes in BW, leptin mRNA, and serum concentrations of IGF-I and leptin normalized to the week of puberty (Exp. 1). Body weight accounted for most of the variation associated with the onset of puberty in the full regression model (R2 = 0.99; P < 0.01). However, serum leptin was closely related to changes in BW (r = 0.85; P < 0.02) and in the absence of BW was most predictive of pubertal onset (r2 = 0.73; P < 0.01). Mean concentrations of leptin increased (P < 0.0001) linearly from 16 wk before until the wk of pubertal ovulation in yearling heifers reaching sexual maturation from early spring to midsummer. Leptin mRNA transformed to a percent of the value at puberty increased (P < 0.02) as puberty approached, but serum leptin and leptin mRNA values were not well correlated. We found no evidence of leptin-binding proteins in serum of developing heifers. Combined mean serum concentrations of IGF-I (ng/mL) during periods III and IV (-9 wk to wk of puberty; 216.6 +/- 9) were 21% higher (P < 0.0001) than combined mean concentrations of IGF-I during periods I and II (-19 to wk of puberty; 193 +/- 10). In mature heifers and cows (Exp. 2), serum leptin tended to decrease (P = 0.10) during the late luteal/early follicular phase of the estrous cycle, which corresponded to a reduction (P < 0.03) in adipocyte leptin gene expression. In mature ovariectomized cows, serum concentrations of leptin increased (P < 0.001) by 34% from early winter to the summer solstice and remained unchanged throughout the remainder of the year (Exp. 3). Results from these studies indicate that marked increases in both circulating leptin and leptin gene expression occur in developing heifers during pubertal development and are associated with increases in serum IGF-I and BW. Seasonal effects on circulating leptin observed in mature cows from winter to summer could also plausibly account for a portion of the prepubertal rise in serum leptin observed in heifers.

Leptin and its role in the central regulation of reproduction in cattle.

Leptin, a 16kDa product of the adipose obese (ob) gene, has been shown to contribute to the regulation of energy metabolism, feeding behavior, and reproduction in several monogastric species, including humans. Recent reports have provided evidence that the leptin gene is functionally relevant in cattle and sheep, and may contribute to an array of important reproductive events, including puberty. Leptin gene expression and circulating leptin increase markedly during sexual maturation in heifers reaching puberty during late spring or early summer. In addition, serum leptin concentrations increased by over 30% from early winter to the summer solstice in mature cows, and also increased with significant changes in adiposity. However, only limited changes in circulating leptin have been observed during the estrous cycle. Short-term fasting of growing peripubertal heifers causes marked reductions in leptin gene expression and circulating leptin, concomitant with declines in LH pulse frequency, and serum concentrations of insulin and IGF-1. Although short-term fasting of mature cows in excellent body condition is without effects on LH pulse frequency, it has remarkably similar metabolic effects to those observed in heifers. Moreover, ICV administration of recombinant oleptin resulted in a marked hypersecretion of LH in fasted cows, and in vitro studies using both hypothalamic and anterior pituitary explants have provided evidence that this effect is at the pituitary level. Paradoxically, ICV administration of oleptin normalized circulating insulin in fasted cows but hleptin was without effect on insulin in estradiol-implanted wethers. Collectively, work in cattle and sheep indicates that leptin can modulate both the hypothalamic-pituitary axis and endocrine pancreas under defined nutritional conditions. Additional work to more fully characterize these roles is clearly warranted and could lead to the development of novel strategies for modifying reproductive potential in food-producing species.

Comparison of three approaches for synchronization of ovulation for timed artificial insemination in Bos indicus-influenced cattle managed on the Texas gulf coast.

Our objectives were to compare the relative efficacies of three protocols designed to synchronize ovulation for timed artificial insemination (AI) of predominantly Brahman-influenced cows and heifers. In Exp. 1, 273 Brahman x Hereford (F1) cows at three locations were stratified by BW, body condition score (BCS), age, and days postpartum and assigned randomly to three treatments: 1) Syncro-Mate-B (SMB), 2) norgestomet-prostaglandin (NP), and 3) Ovsynch. The management goal required that cows have a minimum BCS of 5 and be at least 36 d postpartum (PP) at treatment onset. However, final results included 23 cows (8.4%) whose BCS fell below 5. In Exp. 2, 286 pubertal beef heifers were stratified by BW and BCS and allocated randomly to the three treatments. Heifers were predominantly Brahman crossbred (n = 265; Brahman x Hereford, F1; Santa Cruz) or purebred Brahman-influenced (Santa Gertrudis) with a smaller number (n = 21) of Hereford heifers also included. For both experiments, SMB treatment consisted of a 9-d norgestomet ear implant plus an estradiol valerate/norgestomet injection on d 0. Norgestomet-prostaglandin-treated females were implanted with a SMB implant without the estradiol valerate/norgestomet injection at the time of implant insertion and received 25 mg prostaglandin F2alpha (PGF) i.m. 2 d before implant removal. Ovsynch consisted of 100 microg GnRH i.m. on d 1, 25 mg PGF i.m. on d 8, and a second GnRH injection on d 10. Beginning on d 9, calves were removed for 48 h in Exp. 1. Cattle in SMB and NP groups in both experiments were timed-inseminated 48 to 54 h after implant removal and at 12 to 24 h after the second GnRH injection (Ovsynch). Timed AI conception rates did not differ between the SMB (45.1%) and Ovsynch (42.4%) groups; however, conception rate in the NP group tended (P < 0.12) to be lower overall than in the other groups due to a reduced (P < 0.05) conception rate in cows that were < 60 d PP at treatment onset. Conversely, timed-AI conception was greatest (P < 0.056) in NP (54.7%) compared with SMB (40.4%) and Ovsynch (39.1%) for heifers in Exp. 2. We conclude that in mature, suckled beef cows with Brahman genetic influence, SMB and Ovsynch perform similarly when cow eligibility relies primarily on BCS and minimum days PP. The NP treatment results in lower conception in cows < 60 d PP compared with SMB and Ovsynch. However, in nulliparous Brahman-influenced heifers that are confirmed to be pubertal, NP may be superior to the other two treatments for timed AI.

Central infusion of recombinant ovine leptin normalizes plasma insulin and stimulates a novel hypersecretion of luteinizing hormone after short-term fasting in mature beef cows.

The present studies tested the hypotheses that short-term fasting would reduce leptin gene expression and circulating concentrations of leptin and insulin in mature, ovariectomized, estradiol-implanted cows and that intracerebroventricular infusions of recombinant ovine leptin (oleptin) would attenuate reductions in insulin concentration and stimulate LH secretion. Ovariectomized cows were assigned to either control (normal fed; n = 6) or fasted (60 h of fasting; n = 7) groups and infused with 200 microg recombinant oleptin three times at hourly intervals on Day 2 (n = 6 per group). Fasting decreased plasma concentrations of insulin (P < 0.01) and leptin (P < 0.04) but, as expected, did not reduce plasma concentrations of glucose or any LH secretion variable. Central infusion of leptin on Day 2 increased (P < 0.01) plasma concentrations of leptin in both control and fasted groups. Concomitantly, leptin treatment increased plasma insulin (P < 0.01) and LH (P < 0.03) concentrations in fasted but not in control cows. Increases in overall mean and baseline concentrations of LH after leptin treatment were the result of an augmentation of the size of LH pulses. The effects of fasting on leptin gene expression and the potential diurnal effects on circulating leptin were examined in a group of cows (n = 12) not treated with leptin. Fasting for 60 h reduced (P < 0.001) leptin gene expression by 30%, and no diurnal effects on circulating leptin were observed. These results indicate that although short-term fasting does not reduce the frequency or amplitude of LH pulses or the concentration of LH in mature cows, this nutritional perturbation clearly sensitizes both the hypothalamic-pituitary axis and endocrine pancreas to exogenous leptin, which in these experiments resulted in heightened secretion of both LH and insulin.

Physiological regulation of maternal behavior in heifers: roles of genital stimulation, intracerebral oxytocin release, and ovarian steroids.

We tested the hypotheses that 1) epidural anesthesia at parturition would block both peripheral and central release of oxytocin and eliminate the development of maternal behavior in primiparous heifers and 2) estradiol priming, genital stimulation, and appropriate neonatal stimuli would induce maternal behavior in nulliparous heifers. In experiment 1, primiparous crossbred heifers (n = 13) with cannulas in the third cerebroventricle (IIIV) were assigned randomly to receive epidural treatments of saline (SAL; n = 6) or lidocaine HCl (EPI; n = 7) at the onset of labor induced between Days 270 and 280 of gestation. Epidural anesthesia blocked (P < 0.001) both central and peripheral release of oxytocin and markedly reduced (P < 0.05) or eliminated licking behaviors during a 3-h period following parturition as compared with SAL. Following approximately 1 wk of controlled daily suckling, during which calves were permitted access only to the inguinal region of their dams (three times daily for 10 min each time), a second maternal behavior test was performed. Although licking behavior remained markedly reduced (P < 0.001) in the EPI compared with the SAL groups, all heifers accepted their calf at the udder. In experiments 2-4, neither estradiol priming in ovariectomized heifers nor estradiol plus progesterone in intact heifers resulted in an induction of maternal behaviors following genital stimulation and presentation of a neonate wetted with amniotic fluid. Pelvic sensory deficits apparently block oxytocin release and disturb both short-latency and long-term maternal behaviors but do not result ultimately in rejection of the calf. Combinations of hormonal, sensory, olfactory, and visual cues observed previously to induce maternal behavior in nulliparous ewes do not appear adequate for induction of maternal behavior in nulliparous heifers.

Leptin gene expression, circulating leptin, and luteinizing hormone pulsatility are acutely responsive to short-term fasting in prepubertal heifers: relationships to circulating insulin and insulin-like growth factor I(1).

In the present study, we tested the hypothesis that short-term fasting would reduce leptin gene expression, circulating leptin, and LH pulsatility in prepubertal heifers in association with a decrease in circulating concentrations of insulin and insulin-like growth factor I (IGF-I). Twelve prepubertal crossbred heifers (mean +/- SD = 315 +/- 5 kg body weight) were assigned randomly to one of two treatments in two replicates: 1) control; normal feed consumption (n = 6) and 2) fasted; 48 h of total feed restriction (n = 6). Blood samples were collected at 15-min intervals for 8 h on Days 0 and 2 of the experiment and twice on Day 1. Subcutaneous fat samples were collected before treatment onset (Day -1) and at the end of the intensive blood sampling on Day 2. Acute feed restriction markedly reduced leptin mRNA in adipose tissue (P < 0.01) and circulating concentrations of leptin (P < 0.05), IGF-I (P < 0.01), and insulin (P = 0.05) as compared with controls on Day 2. Moreover, the treatment x day interaction (P < 0.076) and within-day contrasts (expressed as a percentage of Day 0 values) revealed that the mean frequency of LH pulses in the fasted group was lower (P < 0.06) than in controls on Day 2. Neither mean concentrations of growth hormone (GH) nor GH secretory dynamics were affected by acute feed restriction. Fasting-mediated decreases in leptin gene expression and circulating leptin, in association with reductions in secretion of IGF-I, insulin, and LH, provide a basis for investigating leptin as a hormone signaling energy status to the central reproductive axis in cattle.

Distribution and repeatability of anterior pituitary responses to GnRH and relationship of response classification to the postpartum anovulatory interval of beef cows.

Our objectives were to investigate the phenotypic variation in anterior pituitary responsiveness to GnRH (100 microg, i.v.) of beef cows between d 5 and 8 postpartum, estimate repeatability, and determine the relationship between response classification and duration of the postpartum anovulatory interval (PPI). Brahman x Hereford (F1) cows (n = 137) and primiparous heifers (n = 58) were evaluated. Response classifications (Class) included peak LH (Low, Intermediate, or High; Class I) and time to peak LH (Early, 10 to 30 min or Late, 60 to 120 min; Class II). The independent effects of Class I and II on PPI were determined in 145 of 195 cows through twice-weekly serum samples analyzed for progesterone. For Class I, pituitary responses to GnRH approximated a normal distribution and, by definition, differed (P < .001) in magnitudes of peak LH and area under the curve (AUC). For Class II, 111 and 84 cows exhibited early and late peaks, respectively; mean AUC was greater (P < .05) in cows exhibiting late compared with early peaks. Pretreatment LH (P < .01) and estradiol-17beta (P < .004) influenced responses in one or both response classes. Pluriparous cows had shorter (P < .035) PPI than primiparous cows. Class I did not influence the duration of the PPI; however, in Class II, cows with late peaks exhibited an average PPI that was 8 d shorter (P < .025) than in those with an early peak. To estimate repeatability of pituitary responses, 18 classified cows were subsequently rechallenged with GnRH at d 170 of gestation and at the next postpartum period. Although means for each of these challenges differed (P < .05) throughout in both Classes I and II, the small sample size used to make the estimate failed to yield significant (P > .10) interclass correlations. Nevertheless, overall results provide evidence that variability in individual pituitary responses to GnRH could be targeted as a selection marker to improve reproduction.

Injection of neuropeptide Y into the third cerebroventricle differentially influences pituitary secretion of luteinizing hormone and growth hormone in ovariectomized cows.

Hypothalamic neurons that control the luteinizing hormone (LH) and growth hormone (GH) axes are localized in regions that also express neuropeptide Y (NPY). Increased hypothalamic expression of NPY due to diet restriction has been associated with suppressed secretion of LH and enhanced secretion of GH in numerous species. However, these physiological relationships have not been described in cattle. Thus, two studies were conducted to characterize these relationships using ovariectomized (Experiment 1) or ovariectomized estrogen-implanted (Experiment 2) cows. In Experiment 1, four well-nourished, ovariectomized cows received third cerebroventricular (TCV) injections of 50 and 500 micrograms of NPY in a split-plot design. Venous blood was collected at 10-min intervals from -4 hr (pre-injection control period) to +4 hr (postinjection treatment period) relative to TCV injection. NPY suppressed (P < or = 0.04) tonic secretion of LH irrespective of dose and tended to stimulate (P < or = 0.10) an increase in tonic secretion of GH. In Experiment 2, six ovariectomized cows that were well nourished and implanted with estradiol received TCV injections of 0, 50, or 500 micrograms of NPY in a replicated 3 x 3 Latin Square. Both doses of NPY suppressed (P < 0.06) mean concentration of LH relative to the 0-microgram dose. The 50-microgram dose of NPY tended (P < 0.10) to increase the amplitude of GH pulses. In conclusion, TCV injection of NPY suppressed pituitary secretion of LH and simultaneously tended to increase pituitary secretion of GH.

Gonadotropin-releasing hormone secretion into third-ventricle cerebrospinal fluid of cattle: correspondence with the tonic and surge release of luteinizing hormone and its tonic inhibition by suckling and neuropeptide Y.

Objectives of the current studies were to characterize the pattern of GnRH secretion in the cerebrospinal fluid of the bovine third ventricle, determine its correspondence with the tonic and surge release of LH in ovariectomized cows, and examine the dynamics of GnRH pulse generator activity in response to known modulators of LH release (suckling; neuropeptide Y [NPY]). In ovariectomized cows, both tonic release patterns and estradiol-induced surges of GnRH and LH were highly correlated (0.95; p < 0.01). Collectively, LH pulses at the baseline began coincident with (84%) or within one sampling point after (100%) the onset of a GnRH pulse, and all estradiol-induced LH surges were accompanied by corresponding surges of GnRH. A 500- microg dose of NPY caused immediate cessation of LH pulses and lowered (p < 0.001) plasma concentrations of LH for at least 4 h. This corresponded with declines (p < 0.05) in both GnRH pulse amplitude and frequency, but GnRH pulses were completely inhibited for only 1.5-3 h. In intact, anestrous cows, GnRH pulse frequency did not differ before and 48-54 h after weaning on Day 18 postpartum, but concentrations of GnRH (p < 0.05) and amplitudes of GnRH pulses (4 of 7 cows) increased in association with weaning and heightened secretion of LH. We conclude that the study of GnRH secretory dynamics in third-ventricle CSF provides a reasonable approach for examining the activity and regulation of the hypothalamic pulse generator in adult cattle. However, data generated using this approach must be interpreted in their broadest context. Although strong neurally mediated inhibitors of LH pulsatility (suckling; NPY) had robust effects on one or more GnRH secretory characteristics in CSF, only high doses of NPY briefly abolished GnRH pulses. This implies that the GnRH signal received at the hypophyseal portal vessels under these conditions may differ quantitatively or qualitatively from those in CSF, and theoretically would be undetectable or below a biologically effective threshold when LH pulses are absent.

Effect of daily replacement therapy with recombinant bovine somatotropin on somatotropin, insulin-like growth factor I, and onset of puberty in beef heifers immunized against growth hormone-releasing factor.

Two experiments examined whether replacement therapy with recombinantly derived bovine somatotropin (rbST) would induce puberty in heifers that had been actively immunized at 6 mo of age against growth hormone-releasing factor (GRF). Heifers received daily i.m. injections of 25 mg of rbST (Exp. 1, n = 6; Exp. 2, n = 4) or vehicle (VEH; Exp. 1, n = 6; Exp. 2, n = 4) for 56 d. Serum concentrations of somatotropin (ST, nanograms/milliter) were low in all heifers before first injection in Exp. 1 (1.56 +/- .04) and 2 (.95 +/- .03). During treatment, serum ST was greater (P < .01) in rbST than in VEH heifers (75.4 +/- 4.8 vs 2.8 +/- .1 ng/mL, respectively) in both experiments and remained increased through d 57 (32.2 +/- 6.4 vs .90 +/- .01 ng/mL). IN Exp. 1 and 2, concentrations of serum IGF-I were similar in rbST and VEH heifers before treatment, increased (P < .01) 12 h after first rbST, and remained increased (P < .01) through d 57 in rbST heifers. Concentrations of serum insulin (INS) and plasma glucose (GLU) were similar (P > .10) in rbST and VEH heifers before first injection (Exp. 1 and 2). Serum INS (micro-units/milliliter) was greater (P < .01) in rbST (61.7 +/- 3.7 and 36.0 +/- 2.4) than in VEH (12.4 +/- 1.6 and 8.1 +/- 1.0) heifers on d 1 or 2 only, in Exp. 1 and 2, respectively. In Exp. 1, GLU was increased (P < .05) by rbST on d 2 through 57, but only on d 1 in Exp. 2. Proportion of heifers pubertal by d 21 tended to be greater (P < .07) in rbST (3 of 6) than in VEH (0 of 5) heifers in Exp. 1, but not in Exp. 2 (1 of 4 vs 1 of 4, respectively). All heifers in Exp. 1 and 50% of the heifers in Exp. 2 attained puberty by d 56. Daily rbST increased ST, IGF-I, INS, and GLU but did not hasten onset of puberty in heifers immunized against GRF.

Effect of somatotropin and/or equine chorionic gonadotropin on serum and follicular insulin-like growth factor I and insulin-like growth factor binding proteins in cattle.

We investigated the effect of administration of somatotropin (ST) and/or eCG on insulin-like growth factor I (IGF-I) and IGF-binding proteins (IGFBP) in serum and follicular fluid (FFL) of cattle actively immunized against growth hormone-releasing factor (GRF). Cyclic beef cattle, previously immunized against GRF-(1-29)-Gly-Gly-Cys-NH2 conjugated to human serum albumin (synthesized and provided by Hoffmann-LaRoche, Inc., Nutley, NJ; GRFi, n = 31) or to human serum albumin alone (HSAi, n = 26), received (i.m.): 1) 25 mg recombinantly derived methionyl somatotropin (rbST, n = 14; sometribove provided by Monsanto Co., St. Louis, MO); 2) 1100 IU eCG (n = 10); 3) rbST and eCG (rbST-eCG, n = 15); or 4) vehicle (VEH, n = 17) at 0 and 24 h after receiving prostaglandin F2 alpha (PGF2 alpha). Serum samples were collected at 0 and 40 h after PGF2 alpha, and the ovary bearing the largest follicle (DOM) was removed 44.0 +/- 0.5 h after PGF2 alpha; FFL was harvested from DOM and the subordinate follicle (SUB). Before treatment (0 h), GRFi cows had lower serum ST (0.6 +/- 0.2 vs. 2.2 +/- 0.2 ng/ml; p < 0.01) and IGF-I (26 +/- 4 vs. 72 +/- 4 ng/ml; p < 0.01), but greater IGFBP-2 (594 +/- 48 vs. 384 +/- 52 ng/ml; p < 0.01) than HSAi cows. Serum and FFL concentrations of IGF-I or IGFBP-2 were not different between rbST- and rbST-eCG-treated cows or between VEH- and eCG-treated cows at Hour 40 after the initial treatment injection; therefore, data were combined and designated as rbST and VEH, respectively. Serum IGF-I was increased to a greater extent (percentage increase above 0 h) by rbST treatment in GRFi (362 +/- 24) than in HSAi (176 +/- 16) cows (immunization by treatment, p < 0.01). Across GRFi and HSAi, rbST lowered serum IGFBP-2 (342 +/- 31 vs. 541 +/- 27 ng/ml, rbST vs. VEH; p < 0.01). Diameters of DOM or SUB were not affected by immunization or treatment. Concentrations of IGF-I and IGFBP-3 (determined by ligand blot analysis) in FFL from both DOM and SUB were lower (p < 0.05) in GRFi than in HSAi cows. In contrast, IGFBP-2 in FFL was elevated in SUB, but not DOM, in GRFi compared to HSAi cows.(ABSTRACT TRUNCATED AT 400 WORDS)

Changes in serum somatotropin, somatotropin mRNA, and serum and follicular insulin-like growth factor-I in response to feed restriction in cows actively immunized against growth hormone-releasing factor.

Cyclic cows immunized against growth hormone-releasing factor (GRFi, n = 19), human serum albumin (HSAi, n = 10), or not immunized (CON, n = 18) were used to investigate the effects of feed restriction on serum and pituitary somatotropin (ST), pituitary ST mRNA, and serum and follicular IGF-I. Cows were either fed 2.7 kg/d cottonseed hulls (R) or given ad libitum access to feed (AL) for 15 d. Ovaries bearing the largest follicle and pituitaries were collected on d 14, at 44 to 45 h after injection of prostaglandin F2 alpha. Data from CON and HSAi cows were similar; thus, data were combined (represented as CON). Serum ST (nanograms/milliliter) on d 13 was greater (P < .09) in CON-R (5.3) than in CON-AL (3.9), whereas ST in GRFi-AL (1.1) and GRFi-R (1.1; pooled SE = .4) were similar. Hemipituitary weight (grams) and ST mRNA (arbitrary units) were greater (P < .05) in CON (1.5 +/- .1 and 135 +/- 25) than in GRFi (1.0 +/- .1 and 90 +/- 18) cows. Across immunization, ST mRNA and pituitary ST concentration (mg/100 mg of tissue), respectively, were greater (P < .06) in R (152 +/- 22 and 22.5 +/- 1.9) than in AL (73 +/- 16 and 17.3 +/- 1.8) cows. Immunization and diet decreased (P < .05) serum IGF-I (nanograms/milliliter) on d 13 (CON, 176 +/- 7 vs GRFi, 42 +/- 8; AL, 120 +/- 7 vs R, 98 +/- 8). Concentrations of IGF-I in follicular fluid (FFL) from the largest follicle were lower in GRFi (29 +/- 3) than in CON (102 +/- 6) cows; however, IGF-I in FFL was similar in AL (70 +/- 9) and R (71 +/- 10) cows. In conclusion, GRFi decreased serum ST and IGF-I, and decreased ST mRNA. Feed restriction increased serum ST and ST mRNA, and decreased serum IGF-I. Although feed restriction and GRFi decreased serum IGF-I, concentrations of IGF-I in FFL were decreased only by GRFi.

Endocrine events prior to puberty in heifers: role of somatotropin, insulin-like growth factor-I and insulin-like growth factor binding proteins.

We have utilized active immunization against growth hormone releasing factor (GRF) to investigate relationships among somatotropin (ST), insulin-like growth factor-I (IGF-I), IGF binding proteins (IGFBP) and ovarian function in heifers. Active immunization against GRF (GRFi) has been demonstrated to abolish episodic release of ST and decrease serum concentrations of IGF-I. In initial experiments investigating onset of puberty, breeds of heifers differing in growth rate and reproductive traits (Angus, Charolais and Simmental) were immunized against GRF or served as controls (immunized against carrier protein, human serum albumin, HSAi). GRFi decreased rate of muscle and skeletal growth, but increased deposition of adipose tissue. In Angus and Charolais, but not Simmental heifers, GRFi at 6 mo of age significantly delayed onset of puberty beyond 18 mo of age. Retrospective analyses of serum IGF-I revealed that GRFi heifers reaching puberty at a normal age had greater pre-treatment (6 mo of age) IGF-I than GRFi heifers in which puberty was delayed. Collectively, these results strongly indicate that the bovine hypothalamic-hypophyseal-ovarian axis is particularly sensitive to changes in metabolism at or near 6 mo of age. Another series of experiments tested the hypothesis that lowering serum IGF-I via GRFi initially at 3 mo of age would increase the percentage of Angus and Simmental heifers not reaching puberty. Three mo old Angus and Simmental heifers were assigned to GRFi (n = 18), HSAi (n = 14) or received no treatment (controls, n = 16). HSAi and GRFi heifers were unilaterally ovariectomized (ULO) at 6 mo of age. As anticipated, GRFi at a younger age increased percentage of heifers not reaching puberty; over 75% of control and HSAi heifers reached puberty by 14 mo of age compared to 22% of GRFi heifers. Serum and follicular fluid (FFL; follicles < or = 4 mm) concentrations of IGF-I were suppressed by GRFi. Serum, but not FFL concentrations of IGF binding protein-2 (IGFBP-2) were greater in GRFi than in HSAi heifers. GRFi delayed puberty apparently by suppressing follicular growth because number of follicles < or = 7 mm was significantly lower in GRFi than in HSAi heifers. In conclusion, active immunization against GRF at 3 or 6 months of age delays puberty in beef heifers. Delayed puberty was preceded by suppression of follicular growth, and decreased concentrations of IGF-I in serum and follicular fluid.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of exogenous glucose or colostrum on body temperature, plasma glucose, and serum insulin in cold-stressed, newborn Brahman calves.

The effects of ambient temperature and source of exogenous energy (glucose or colostrum) on the ability of newborn Brahman calves to maintain rectal temperature (RT) were determined. All calves were removed from dams within 30 min of birth, before suckling. Calves were catheterized and placed in either a warm (25 degrees C) or cold (5 degrees C) environment for 150 min and given either colostrum or glucose. This resulted in four groups (warm colostrum, n = 7; cold colostrum, n = 7; warm glucose, n = 6; cold glucose, n = 6). Blood samples and RT were obtained at 15-min intervals during warm or cold through 150 min, when calves were removed from cold, and at 180, 240, and 300 min. After 60 min, each calf was given either 1 L of colostrum (38 degrees C) from its dam or glucose (38 degrees C) infusion of 750 mg/kg BW. Plasma glucose concentrations were determined by enzymatic techniques and serum insulin concentrations by RIA. Calves exposed to cold or warm air temperatures had similar declines in rectal temperature from 0 to 60 min. Colostrum-fed, cold calves had a greater (P less than .07) decrease in RT than did colostrum-fed, warm calves from 75 through 150 min; glucose-infused warm and cold calves had intermediate decreases in RT. Plasma glucose increased (P less than .0001) in glucose-infused compared with colostrum-fed calves at 75 min, but glucose-infused calves had lower (P less than .02) glucose levels from 180 to 300 min. Higher (P less than .05) glucose concentrations