Effects of passive immunization of growing guinea-pigs with an insulin-like growth factor-I monoclonal antibody.

The physiological importance of circulating as opposed to locally produced insulin-like growth factor-I (IGF-I) has not been determined. By using a passive immunoneutralization technique, our objectives were to evaluate the role of circulating IGF-I in the regulation of animal growth and pituitary GH content. A monoclonal antibody (MAb) to IGF-I, generated in our laboratory, has an affinity (Ka) of 0.13 litres/pmol for recombinant human IGF-I (rhIGF-I). Cross-reactivities of recombinant des-tripeptide IGF-I and recombinant bovine IGF-II were approximately 40 and 8% respectively. This MAb inhibited binding of purified hIGF-I to human placental membranes. In a radioimmunoassay based on displacement of 125I-labelled rhIGF-I from the MAb, displacement curves generated with dilutions of acid-gel chromatography extracts of guinea-pig serum and rhIGF-I standards were parallel. Twenty-four, 3-week-old male guinea-pigs were treated with the IGF-I MAb, a bovine herpes virus-I (BHV-I) MAb (control MAb) or vehicle (phosphate-buffered saline) (n = 8 per group). Treatments were administered i.p. every 3 days for 24 days at a dose of 20 mg/kg body weight. Blood was obtained on day 23 (48 h after treatment) and on day 25 (24 h after treatment). In a liquid-phase assay, serum from the IGF-I MAb-treated group bound 38 +/- 8% (mean +/- S.E.M.) (day 23) and 56 +/- 7% (day 25) of an 125I-labelled rhIGF-I trace at a final dilution of 1:10,000. Because of the development of an anti-mouse immune response in the guinea-pigs, these parameters would probably have been much greater during the first 2 weeks of the trial. Of the total IGF-I in serum, 50 +/- 5% and 61 +/- 4% could be immunoprecipitated with an excess of rabbit anti-mouse immunoglobulin in samples from days 23 and 25 respectively. Comparisons between the groups treated with IGF-I MAb and BHV-I MAb revealed no significant differences in whole animal growth rate, growth of individual tissues, or pituitary GH content. Mean serum concentrations of IGF-I were 69 and 99% greater in the IGF-I MAb-treated group than in the BHV-I MAb-treated group on days 23 and 25 respectively. These differences probably resulted from an extension of the half-life of IGF-I in serum of animals treated with the IGF-I MAb.(ABSTRACT TRUNCATED AT 400 WORDS)

Stimulation effect of prostaglandin F2ALPHA on the plasma concentration of testosterone in bulls.

The concentrations of peripheral plasma testosterone were measured by radioimmunoassay in samples collected from five bulls, each given i.m. injections of 0, 5. 15, 30 and 60 mg prostaglandin F2alpha (PGF2alpha). Synchronized peaks in testosterone concentration occurred with maximum values 1.2h after treatment. These increases of testosterone persisted significantly longer than those observed to occur as natural episodic peaks during two 24 h periods in the same bulls. The mean peak testosterone concentration after PGF2alpha injection was related to the dose of PGF2alpha, values after 60 and 30 mg doses being significantly greater than after 15 mg. The response produced by a 5 mg dose was not significant. The results indicate that intramuscular injection of PGF2alpha, acutely stimulates synthesis and release of testosterone in fulls.

Effects of progesterone and weaning on LH and FSH responses to naloxone in postpartum beef cows.

Two experiments were conducted to evaluate the effects of naloxone, an endogenous opioid receptor antagonist, on LH and FSH secretion in postpartum beef cows. In Experiment 1, 24 cows were divided into three equal groups. On day 15 postpartum, all cows were bled for 8 hr at 10 min intervals to evaluate LH secretory parameters. On day 18 postpartum, three treatments were administered: (a) saline at 0730 and 1130 hr; (b) 275 mg naloxone at 0730 and 1130 hr; (c) naloxone as in (b) above, plus this group was also treated with 50 mg progesterone (P4) twice daily from day 16 to day 19. In each treatment, jugular vein samples were collected at 10 min intervals from 0800 to 1600 hr. On day 19 the same treatments were administered at the same times, however, all cows were given 25 micrograms GnRH at 1200 hr to evaluate the LH secretory response. Naloxone increased mean LH concentration (P less than .05) and tended to increase pulse amplitude and frequency compared to controls. However, the most dramatic difference was due to P4 treatment which suppressed mean LH, pulse amplitude and frequency. Treatments had no effect on LH secretion in response to a 25 micrograms dose of GnRH. In Experiment 2, the effects of suckling on the naloxone response were examined in 16 postpartum cows. On day 21 postpartum, blood was collected at 10 min intervals for 8 hr and then calves were removed from half the cows. After 3 days of calf removal, all cows were sampled at 10 min intervals for 4 hr; then naloxone was injected after each 10 min sample at a dose rate of 200 mg/hr (33 mg per injection). Naloxone treatment and sampling continued for an additional 8 hr. Calf removal alone had very little effect on LH pulsatility. However, naloxone resulted in increased pulse frequency and mean LH compared to the control period. We conclude that LH release in the early postpartum cow is partially regulated by endogenous opioid peptides. We were unable to detect any effects on FSH secretion nor on pituitary sensitivity to exogenous GnRH.

FSH receptors in the bovine corpus luteum.

Binding of follicle stimulating hormone (FSH) to a crude membrane fraction of bovine corpus luteum (CL) has been detected. This binding meets the usual criteria for a receptor based on specificity, time course of reaction and association constant (Ka = 8.5 x 10(10)M(-1)). Physiological studies with CL removed from heifers at specific times after estrus indicate that day-6 CL had the highest FSH binding. However, a correlation with physiological function was not obvious since some functional mid-cycle CL were high in progesterone and luteinizing hormone (LH) receptor but had nondetectable FSH receptor. Conversely, some late-cycle CL had low progesterone and LH receptor but significant quantities of FSH receptor.

Influence of GnRH infusion on endocrine parameters and duration of postpartum anestrus in beef cows.

The effect of an intravenous infusion of gonadotrophin releasing hormone (GnRH) on the duration of postpartum anestrus in suckled beef cows was studied. Twenty-eight, mature, suckled beef cows were assigned in equal numbers to one of four treatment groups which were based on infusion with saline or GnRH (15ug/hour for 12 hours) and stage postpartum (pp) (20 or 35 days). Serum LH and progesterone were determined by radioimmunoassay for the period which began 5 days pre-infusion and ended at 55 days postpartum (ie: 35 or 20 days post-infusion). Serum LH remained below 5ng/ml during infusion in all control cows. Peak serum LH values, times of LH peaks, and duration of LH responses (means +/- SE) during infusion were 49 +/- 12 ng/ml, 162 +/- 42 minutes and 7.8 +/- 1.3 hours for the 20 day group and 44 +/- ng/ml, 144 +/- 6 minutes, and 8.2 +/- 1.1 hours for the 35 day group respectively. Serum progesterone levels indicated that the proportion of cows showing the onset of estrous cycles within 10 days of infusion was greater in the 20 day pp GnRH group (4/7) than the 20 day pp saline group (0/7) (p < .05) but was not significantly different between the 35 day pp GnRH (4/7) and 35 day pp saline (2/6) groups. The incidence of estrus was not affected by GnRH treatment and was 37% in all cows prior to 55 days pp. It was concluded that infusions of GnRH for 12 hours at a rate of 15 ug/hour could induce estrous cycles in suckled beef cows treated at 20 days postpartum.

Follicular phase gonadotropin secretion in cyclic postpartum beef cows with phlorizin-induced hypoglycemia.

Twenty mature, lactating Hereford-cross cows were used to determine the effect of phlorizin-induced hypoglycemia on gonadotropin secretion following prostaglandin-induced luteolysis. Cows were 43 to 108 d postpartum and had a functional corpus luteum (CL) at the start of infusion treatment (d 1). Infusions consisted of either saline (control) or 3 g/d of phlorizin infused continuously from the time of prostaglandin injection at 1000 on d 1 until 0800 on d 5. Blood samples were collected for determination of plasma concentrations of insulin, glucose and free fatty acids (FFA) and for serum concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and progesterone. Plasma concentrations of insulin (P less than .05) and glucose (P less than .05) were lower, whereas FFA concentrations increased (day X treatment, P less than .05) over the infusion period in phlorizin-treated cows compared with saline-infused controls. Mean serum concentrations of LH (1.17 +/- .10 vs 1.53 +/- .20 ng/ml; P less than .05) and LH pulse amplitude (1.69 +/- .14 vs 2.47 +/- .37 ng/ml; P less than .10) were lower in phlorizin-infused compared with saline-infused cows during the 0 to 24-h period immediately preceding the ovulatory gonadotropin surge. The FSH pulse frequency increased (.33 +/- .11 to .55 +/- .12 pulses/h) in saline-infused cows, but decreased (.61 +/- .10 to .41 +/- .11 pulses/h) in phlorizin-infused cows before the gonadotropin surge. Other characteristics of gonadotropin secretion were similar among phlorizin-infused and saline-infused cows. All but one phlorizin-infused cow ovulated and formed functional CL similar to controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Hypoglycemia alters pulsatile luteinizing hormone secretion in the postpartum beef cow.

This study tested the hypothesis that the increased glucose requirement of lactation had effects that were independent of the suckling-dependent inhibition of postpartum endocrine function in beef cows. Mature Hereford cows were either suckled ad libitum and infused with saline iv (n = 9) from d 2 through 4 (d 0 = jugular catherization on d 32 +/- 3 postpartum); were nonsuckled and infused with saline from d 2 through 4 (n = 10); or were nonsuckled and infused with phlorizin (3 g/d) from d 2 through 4 (n = 10). Nonsuckled cows infused with phlorizin had lower (P less than .05) plasma concentrations of glucose and amino acid nitrogen (AAN) on d 2 compared with pre-infusion levels (d 1), but their metabolic profile returned to levels similar to the suckled cows by d 3 and 4. Nonsuckled cows infused with saline had elevated glucose and insulin and lower AAN and free fatty acids (FFA) on d 3 and 4 compared with pre-weaning (d 1) levels (P less than .05). Nonsuckled cows infused with phlorizin did not show this weaning-induced elevation in glucose and insulin. The number of luteinizing hormone (LH) pulses was not affected by treatment. However, in contrast to the large LH pulses observed in the nonsuckled cows infused with saline, both the suckled cows and the nonsuckled cows treated with phlorizin had more small and fewer large amplitude pulses (P less than .01). Treatment did not affect serum concentrations of follicle stimulating hormone, gonadotropin release in response to gonadotropin releasing hormone (25 micrograms) or the number of cows ovulating by 55 d after calving. We conclude that the increased glucose clearance caused by phlorizin infusion or lactation results in depression of LH pulse amplitude in suckled postpartum beef cows.

Insulin-like growth factor I in follicular development and function in postpartum beef cows.

The objective of this study was to determine whether nutrition affects follicular growth and(or) steroid and insulin-like growth factor I (IGF-I) concentrations in follicular fluid. Beginning 6 d after calving, Hereford-cross cows (n = 28) were fed either 14 (ad libitum) or 7 (restricted) kg.animal1.d-1 of chopped alfalfa-brome hay. Half the cows in each treatment were ovariectomized on d 20 (OVX-20) and the remaining half on d 35 (OVX-35) postpartum. Cow weight and condition score were recorded weekly, and blood was collected thrice weekly for determination of insulin, IGF-I, glucose, and free fatty acid (FFA) concentrations. At ovariectomy, follicular fluid from each follicle greater than or equal to 4 mm in diameter was aspirated for determination of IGF-I, progesterone (P4), and estradiol-17 beta (E2) concentrations. Restricted cows lost more weight after calving than did ad libitum cows (P less than .0001), although all cows lost similar amounts of body condition (time postpartum, P = .008). Concentrations of FFA were elevated (P less than .0001) in restricted cows from wk 2 through 5 after calving but did not change with time in ad libitum cows. Plasma concentrations of glucose were lower in restricted than in ad libitum cows (59.6 +/- .4 vs 61.8 +/- .4 mg/dl; P = .05), but insulin and IGF-I were similar (P greater than .10) between dietary treatments.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in metabolic and reproductive characteristics associated with lactation and glucose infusion in the postpartum ewe.

This study examined mechanisms whereby the metabolic environment interacts with basic reproductive function. Ewes lambing during the breeding season were fed to maintain (MAINT, n = 10) or gain (GAIN, n = 11) body weight during the last 4 mo of gestation. From d 7 to 22 postpartum, ewes were infused iv with saline (n = 10) or glucose at a rate calculated to increase normal glucose entry rate by 75% (n = 11). Blood samples were collected daily to determine plasma concentrations of nutritive metabolites and insulin and at frequent intervals on d 14 and 21 to determine serum gonadotropin concentrations. Hypothalami and pituitaries were collected on d 22 to determine hormone content and receptor concentrations. Plasma concentrations of nutritive metabolites and insulin indicated that MAINT ewes mobilized more (P less than .01) body fat and protein reserves during gestation and early lactation than ewes in the GAIN group. Glucose infusion elevated plasma concentrations of glucose (P less than .05) and insulin (P less than .07) and reduced (P less than .05) fat and protein mobilization, even though it depressed feed intake (P less than .001), compared with saline infusion. Hypothalamic gonadotropin-releasing hormone (GnRH), pituitary luteinizing hormone (LH) and follicle-stimulating hormone (FSH) content and pituitary GnRH receptor concentration were similar between treatments.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum concentrations of IGF-I in postpartum beef cows.

Four experiments assessed changes in serum IGF-I under various physiologic conditions in postpartum cows. In Exp. 1, anestrous suckled cows (n = 25) were infused for 6 d with either saline or glucose at two different infusion rates. In Exp. 2, anestrous cows (n = 29) received either a saline (weaned and suckled controls) or 3 g/d phlorizin (weaned phlorizin) infusion for 3 d. Calves from the weaned groups were removed from 15 h before and throughout infusions. In Exp. 3, cycling suckled cows (n = 20) received prostaglandin F2 alpha (PGF2 alpha) when the 5-d saline or phlorizin infusion began. In Exp. 4, suckled cows (n = 20) had ad libitum access to feed or received 50% of control feed consumption from 30 to 40 d postpartum. Increasing glucose availability (Exp. 1) increased (P less than .05) serum IGF-I by 30 to 35%. IGF-I remained stable after weaning (Exp. 2) in phlorizin-infused cows (128.8 +/- 12.7 ng/ml), but increased (P less than .05) by 3 d after calf removal in weaned control cows (152.2 +/- 7.5 ng/ml). IGF-I also remained stable in phlorizin-infused cows following PGF2 alpha injection (Exp. 3), but increased in control cows by 2 d after PGF2 alpha (156.8 +/- 18.3 on d 2 vs. 133.7 +/- 9.8 ng/ml pre-injection; P less than .05) and remained elevated (P less than .05) during the periovulatory period. In cows receiving restricted feed intake (Exp. 4), IGF-I decreased by approximately 50% within 4 d of feed restriction (71.3 +/- 9.4 vs 137.4 +/- 16.6 ng/ml; P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Endocrinology of the postpartum period in the Pelibuey ewe.

The postpartum (PP) period in the Pelibuey ewe was studied. Laparotomies were performed on 14 ewes in the first year at d 10, 20 and 30 PP, and at d 10 and 20 PP in the second year on 17 ewes. Progesterone concentrations were determined in serum taken daily, from 4 to 7 d after parturition until estrus. Temporal fluctuation of luteinizing hormone (LH) was determined in samples taken at 30-min intervals for 4 h weekly. The mean interval from lambing to first ovulation was longer (P less than .001) in 1980 (59 +/- 4.9 d) than 1979 (26 +/- 3.1 d), the mean interval from lambing to first estrus was also longer (P less than .001) in 1980 (91 +/- 5.6 d) than 1979 (51 +/- 5.5 d). Follicles were present on the ovaries of the majority of the ewes at d 10. The mean diameter of the largest follicles on each ovary was reduced (P less than .025) in ewes in 1980 (6 mm) compared with 1979 (7.7 mm). Corpora lutea (CL) occurred in 67 and 75% of the ewes by d 20 and 30, respectively in 1979; no CL were found by d 20 in 1980. Progesterone profiles suggested that the PP period was composed of a period of anestrus, and a period of cyclic ovarian activity with one, two or three ovulations without behavioral estrus. In some ewes, the first cycle was of shorter duration, and its CL secreted less progesterone (P less than .05) relative to CL of silent and regular estrous cycles. Luteinizing hormone peaks were recorded as early as 6 d PP. When progesterone concentrations were elevated to luteal phase levels, the frequency, but not magnitude, of LH peaks per 4-h bleeding period was reduced (P less than .05) relative to anestrus. It is concluded that there are periods of anestrus and of silent cycles, which precede the first postpartum estrus in Pelibuey ewes.

Termination of pregnancy with cloprostenol and dexamethasone in intact or ovariectomized cows.

Termination of pregnancy in cows was investigated using sham-operated (SH) or ovariectomized (OV) cows treated with either a saline vehicle (V), cloprostenol (PG), dexamethasone (DEX) or dexamethasone and cloprostenol (DEX+PG). Surgery was done at 210 days of pregnancy and treatment was administered 72 hours later. Days (mean+/-S.E.) from treatment to termination of pregnancy for the treatment groups were: sham-operated +vehicle (SH+V): 61.5+/-11.3; ovariectomized+vehicle (OV+V): 53.4+/-15.7; sham-operated+cloprostenol (SH+PG): 61.8+/-1.7; ovariectomized+cloprostenol (OV+PG): 54.5+/-13.1; shamoperated+dexamethasone (SH+DEX): 74.8+/-4.8; ovariectomized+dexamethasone (OV+DEX): 2.8+/-0.4; shamoperated+dexamethasone+cloprostenol (SH+DEX+PG) 26.0+/-23.0; ovariectomized+dexamethasone+cloprostenol (OV+DEX+PG): 7.2+/-4.9. Pregnancies in the OV+DEX and OV+DEX+PG groups were terminated significantly earlier than in all other groups (P<0.05) except the SH+DEX+PG group. These findings suggest that dexamethasone will terminate pregnancy in cows near seven months of gestation after the ovarian source of progesterone has been removed by either an injection of prostaglandin or by ovariectomy.

Induction of abortion in feedlot heifers with a combination of cloprostenol and dexamethasone.

A field trial was conducted to assess the efficacy of a combined prostaglandin F(2)alpha analogue (cloprostenol) and dexamethasone treatment as an abortifacient in feedlot heifers. Heifers were grouped according to stage of gestation as follows: Group I, one to four months, n = 37: group II, four to six months, n = 40: group III, six to eight months, n = 40: group IV, one to eight months, n = 29. Heifers in groups I, II and III received a simultaneous intramuscular injection of 500 mug cloprostenol and 25 mg dexamethasone at the time of rectal palpation for pregnancy diagnosis. Heifers in group IV were subjected to rectal palpation for pregnancy diagnosis but received no treatments. Heifers were observed daily for two weeks for abortion and rectal palpations were done 50 days after treatment to determine reproductive status. HEIFERS ABORTING AFTER TREATMENT WERE AS FOLLOWS: Group I, 37/37; group II, 37/40, group III, 37/40; group IV, 0/29. In each of groups II and III there was one pregnancy and two cases of fetal mummification. The numbers of abortions in groups I, II and III were significantly different from that in group IV (P