Induction of pulsatile LH release, FSH release and ovulation in post-partum acyclic beef cows by repeated small doses of Gn-RH

Five acyclic spring-calving beef cows (20-40 days post partum) were bled every 15 min for 60 h and thereafter every 6 h for 5 days. Gn-RH (5 micrograms) was injected every 2 h for 48 h, starting 12 h after sampling began. Pulsatile patterns of LH release occurred synchronously in response to injection and 4 of the 5 treated animals subsequently ovulated and completed at least one full ovarian cycle. Four of 6 similar control cows were bled every 10 min for 8-h periods at equivalent times post partum. Pulses of LH were seen after approximately Day 25 post partum with a mean pulse frequency of 0.5 per h. There was little evidence of a discrete pulsatile mode of FSH release in any of the treated or control cows. The time to the first significant progesterone rise in the 4 treated and ovulating cows (34.5 +/- 5.6 days post partum) was significantly shorter (P less than 0.05) than in the 6 control animals (66.3 +/- 11.4 days).     

Response of cyclic and post-partum suckled cows to injections of synthetic LH-RH.

Doses of 125, 250 or 500 micrograms LH-RH were injected i.m. into suckled beef cows on approximately Day 11 of an oestrous cycle synchronized by prostaglandin treatment. There was a positive linear relationship between dose of LH-RH and the area under the measured LH peak. Administration of 500 micrograms LH-RH as a single injection to suckled cows 13-32 days post partum resulted in LH release but failed to induce normal ovarian activity. A small transient rise in plasma progesterone for 6--9 days occurred at the expected time after injection in 50% of animals. Administration of 500 micrograms LH-RH to suckled beef cows approximately 20--30 days post partum and a second injection approximately 10 days later at the time when the resulting transient rise in plasma progesterone had returned to basal values induced normal cyclic activity (as shown by progesterone concentrations and observed oestrus) at 35 days compared with 70 days for untreated controls. Pituitary responsiveness to LH-RH, as assessed by LH levels, was found to increase up to 20 days post partum.     

Plasma progesterone and gonadotrophin concentrations and ovarian activity in post-partum dairy cows

Plasma progesterone concentrations in jugular vein blood samples collected every other day after calving from 13 Friesian dairy cows indicated that ovarian cyclic activity was initiated by 16.6 +/- 1.1 (s.e.m.) days post partum, except for 1 cow which did not resume cyclic activity until Day 98 post partum. Rectal palpation of the ovaries indicated that a developing follicle was recognizable at a mean time of 15.7 +/- 2.0 days after calving. During the first oestrous cycle after parturition there was a significantly shorter period when plasma progesterone levels were elevated than during the next 2 cycles. Concentrations of progesterone, LH, FSH and prolactin were determined for 4 cows, in blood samples taken every 6 h from 2 to 36 days post partum. Tonic LH release was lower during the first 10 days than subsequently, but the lack of change in pattern for FSH suggests dissimilar control mechanisms for these hormones during this time. Three cows showed evidence of a resumption of ovarian cyclicity during the sampling period: in 2 there was an initial LH surge of a magnitude which would normally give rise to ovulation, followed 4 days later by an increase in plasma progesterone lasting only 5 and 9 days. This progesterone was considered to be of follicular origin. A second LH surge was followed by the presence of a corpus luteum.     

Hormone responses to exogenous GnRH pulses in post-partum dairy cows

Nine Friesian dairy cows were treated with 2.5 micrograms GnRH i.v. at 2-h intervals for 48 h commencing between Days 3 and 8 post partum. Hormone concentrations were measured in jugular venous plasma. An episodic pattern of LH release was induced in all animals and there was no significant change in amplitude during treatment. However, cows treated between Days 7 and 8 ('late') showed higher LH episode peaks than did those treated between Days 3 and 6 ('early'). Plasma FSH concentrations showed a less clear episodic pattern in response to GnRH injection. The mean height of FSH responses to GnRH tended to be higher in the 'early' group than in the 'late' group, as did mean FSH concentrations during the pretreatment sampling period. Although clear episodic changes were not observed, GnRH treatment induced a rapid sustained rise in plasma oestradiol-17 beta concentrations, indicating the responsiveness of ovarian follicles to gonadotrophin stimulation early in the post-partum period. There was no difference in oestradiol-17 beta concentrations between the 'early' and 'late' groups during the treatment period. Only one cow exhibited preovulatory-type LH, FSH and oestradiol-17 beta surges during the 96-h post-treatment sampling period. It is concluded that: (1) responsiveness to GnRH pulses increases significantly and FSH responsiveness tends to decrease with time post partum, (2) ovarian follicles are able to secrete oestradiol-17 beta in response to GnRH-induced LH and FSH release during the early post-partum period and there is no time-dependent change in responsiveness; and (3) the lack of preovulatory surges, except in one cow, may reflect a temporary defect in the positive-feedback mechanism by which high concentrations of oestradiol-17 beta induce preovulatory gonadotrophin release.     

The effect of exogenous progesterone on plasma LH and milk progesterone concentrations in multiple-suckling post-partum cows

Fourteen Friesian cows each suckling four calves were treated for a 7-day period (a) between days 20–40 post partum with progesterone-releasing intravaginal devices (PRID) containing 2% progesterone (Group 1; n = 5), (b) between days 51–264 post partum with PRIDS containing 2% progesterone (Group 2; n = 6) and (c) between days 29–214 days post partum with PRIDS containing 0% progesterone (Group 3; n = 3). Mean plasma LH concentrations decreased during PRID treatment in Group 2 cows only and pre-ovulatory LH surges were observed in $\text{5}\text{6}$ of these cows between 38 and 84 h after coil removal. All Group 2 cows underwent at least one ovarian cycle following PRID removal. No pre-ovulatory LH surges were observed in either Group 1 or Group 3 cows and only one cow (Group 3) underwent an ovarian cycle after treatment. It is suggested that there is an increase in pituitary responsiveness to the feedback effects of progesterone during the post-partum period.     

Plasma concentrations of FSH and LH in entire and castrated prepubertal bull calves treated with Gn-RH.

In bulls there was no increase in plasma FSH and only a small increase in LH over the first 14 weeks of age. In steers (castrated) plasma LH and FSH were unchanged for the first 3 weeks but increased significantly at 7 and 14 weeks. After 100 micrograms Gn-RH, LH release in bulls was minimal until 7 and 14 weeks and there was no comparable rise for FSH. LH and FSH responded to Gn-RH throughout the trial in the steers. The neonatal calf testes selectively inhibited the release of FSH from the pituitary even when challenged with Gn-RH.     

Effect of endogenous and exogenous progesterone on the oestradiol-induced LH surge in dairy cows

Four cows released an LH surge after 1.0 mg oestradiol benzoate administered i.m. during the post-partum anoestrous period with continuing low plasma progesterone. A similar response occurred in the early follicular phase when plasma progesterone concentration at the time of injection was less than 0.5 ng/ml. Cows treated with a progesterone-releasing intravaginal device (PRID) for 8 days were injected with cloprostenol on the 5th day to remove any endogenous source of progesterone. Oestradiol was injected on the 7th day when the plasma progesterone concentration from the PRID was between 0.7 and 1.5 ng/ml. No LH surge occurred. Similarly, oestradiol benzoate injected in the luteal phase of 3 cows (0.9-2.1 ng progesterone/ml plasma) did not provoke an LH surge. An oestradiol challenge given to 3 cows 6 days after ovariectomy induced a normal LH surge in each cow. However, when oestradiol treatment was repeated on the 7th day of PRID treatment, none released LH. It is concluded that ovaries are not necessary for progesterone to inhibit the release of LH, and cows with plasma progesterone concentrations greater than 0.5 ng/ml, whether endogenous or exogenous, did not release LH in response to oestradiol.     

Modulation of functional capacity of small ovarian follicles in the post-partum cow by prolactin

This study was undertaken to examine the possibility that the prolonged anovulatory period frequently experienced by the post-partum cow is due to a disruption of function at the ovarian level promoted by the high, suckling-induced, blood prolactin concentrations. Fifteen cows, less than 35 days post partum, were allocated to three groups (1, 3 and 5) and given no hormonal treatment, prostaglandin plus pregnant mare serum gonadotrophin (PMSG) treatment or injected with 2-bromo-alpha-ergocryptine to reduce circulating prolactin levels. Ten synchronized cyclic cows were allocated to two groups (2 and 4) and given prostaglandin or prostaglandin plus PMGS treatment. All cows were ovariectomized 1 or 2 days after treatment of Graafian follicles less than 9 mm in diameter were selected after dissection from the ovaries. The follicles were cultured for 18 h with or without prolactin (1 microgram/ml) and steroid accumulation in the culture medium estimated. The follicles were then separated into theca and granulosa which were incubated for 40 min with LH (1 microgram/ml) or FSH (5 micrograms/ml). Cyclic AMP concentrations were estimated as an indication of tissue responsiveness to gonadotrophins. The secretion of oestradiol-17 beta, progesterone, testosterone or androstenedione during 18 h culture did not differ between follicles isolated from post-partum or cyclic cows. The presence of prolactin in the culture medium had no overall effect on steroid secretion although some specific effects within each group were noticed. Incubation with LH increased cyclic AMP levels in the theca but the granulosa did not respond. Likewise FSH increased cyclic AMP levels in granulosa preparations but not in theca. There were no differences in response between post-partum and cyclic cows, but exposure of the follicles to prolactin in vitro did significantly reduce the LH-induced increase in cyclic AMP levels in isolated theca. We have concluded that endogenous prolactin may modify but does not inhibit the resumption of ovarian function following parturition in the beef cow.     

Assessment of the sexual and antler potential of the male white-tailed deer (Odocoileus virginianus) by Gn-RH stimulation test

Twelve mature white-tailed bucks were injected with gonadotropin regulating hormone (Gn-RH, 100 micrograms/deer) during the rut (November) and during the spring (April). In the rut, superior bucks (with actual or potential large body weight, trophy antlers and a high social rank) responded to Gn-RH with a small increase of LH (below 20 micrograms/ml) and a profound rise in testosterone (T) (30-50 ng/ml). The inferior animals exhibited high increase of LH (30-40 ng/ml) but a low rise in T (below 10 ng/ml). FSH levels increased only slightly after Gn-RH and the concentrations were not related to reproductive performance. During the spring, increase in LH levels after Gn-RH administration greatly exceeded the rise of T, but no relationship was found between hormonal levels and the reproductive potential. FSH levels increased remarkably after Gn-RH administration. Gn-RH (administered during the rut) might be used for assessment of the potential for reproductive and antler performance.     

Alterations in hypophysial responsiveness to synthetic GnRH at different postpartum intervals in Murrah buffalo (Bubalus bubalis )

The objective of this study was to investigate the hypophysial responsiveness to GnRH at different intervals post partum in Murrah buffalo. Plasma LH and FSH levels were measured at 1 h before and upto 6 h subsequent to the administration of GnRH (1 ug/kg body weight) or saline on Days 2, 20 and 35 post partum in 2 groups of buffalo (n=4 each). Plasma progesterone levels were measured in samples collected once daily from Day 3 to Day 46 post partum. Pretreatment basal LH levels exhibited a progressive increase from Day 2 through Day 35 post partum, while the basal FSH levels increased only until Day 20 post partum. Following a highly subdued LH response to GnRH on Day 2 post partum, a 408% increase (P < 0.01) was observed in the total LH released in response to GnRH on Day 20 post partum, followed by a 20% reduction (non-significant) over Days 20 to 35 post partum. The interval from parturition was highly correlated with total LH released (r = 0.711, P < 0.01). Unlike LH, a substantial amount of FSH was released following GnRH treatment on Day 2 post partum, which was not significantly different from the FSH response on Days 20 and 35 post partum. The LH and FSH response to GnRH was not significantly different between animals in which luteal activity resumed and in those which showed no luteal activity post partum. While pointing to a dramatic enhancement in the hypophysial responsiveness to GnRH between Days 2 and 20 post partum, these results suggest that pituitary responsiveness to GnRH does not appear to be the limiting factor for resumption of estrous cycles by Day 35 post partum in Murrah buffalo.     

Pattern of luteinizing hormone (LH) and follicle stimulating hormone (FSH) in bovine blood plasma after injection of a synthetic gonadotropin-releasing hormone (Gn-RH)

A synthetic gonadotropin-releasing hormone (Gn-RH) was administered to female and male adult bovines in order to study the release of luteinizing and follicle-stimulating hormones into blood by the pituitary gland. Plasma LH and FSH were determined by means of a radioimmunological method. In females as well as in males, increasing doses of Gn-RH (range 50 to 1500 μg) administered i.v. or i.m. caused a linear increase in plasma LH. The release of FSH evidently was curvilinear over the same dosage range.After 2 or 3 injections of Gn-RH every 3 hours, or every 24 hours or more, smaller amounts of LH were released; repeated treatment did not result in reduction of FSH. Thus pituitary depletion of LH occurred more readily than FSH. The effect of Gn-RH on plasma levels of LH and FSH at various stages of the estrous cycle shows a tendency for an increasing release of both gonadotropins on Days 17 – 18 in comparison to Days 4 – 5 or Days 11 – 12.The results suggest that, within the limits allowed by the heterogenous FSH assay and the method of administration used in these experiments, synthetic Gn-RH does not evoke completely normal physiological responses. Therefore, further work is needed to determine its role in improving reproductive function.     

Role of the corpus luteum of pregnancy in controlling pituitary gonadotrophin secretion during the early post-partum period in the ewe

No difference was found between 5 intact ewes and 5 ewes from which the CL had been excised at Day 70 of pregnancy in the plasma concentration of progesterone at Day 140, and concentrations of progesterone remained below 0.2 ng/ml during the first 20 days post partum. Plasma concentrations of LH, frequency and amplitude of LH pulses were low at Day 140 and increased considerably, particularly in the CL-excised ewes, as early as Day 5 post partum. No significant differences were found between the two groups of ewes in the mean plasma concentrations of FSH for any of the 5 stages examined. Taken together, these results suggest that some factor, other than progesterone, associated with the CL of pregnancy is involved in the inhibition of pulsatile LH secretion during the early post-partum period.     

Effect of changes in FSH induced by bovine follicular fluid and FSH infusion in the preovulatory phase on subsequent ovulation rate and corpus luteum function in the ewe

Treatment of Damline ewes with i.v. injections of various doses (2, 5 or 10 ml) of bovine follicular fluid for 72 h after prostaglandin-induced luteal regression resulted in a significant decrease in plasma concentrations of FSH after a 1.5-2 h delay but did not affect LH. The half life of this decrease in plasma FSH levels (156 min) after injection of follicular fluid was similar to that for clearance (159 min) of ovine FSH after infusion. A significant rebound increase in plasma FSH levels occurred by 13 h after all follicular fluid injections, and the magnitude of this rebound was inversely related to the dose of follicular fluid injected. A significant delay in the onset of oestrus occurred only with 5 and 10 ml bovine follicular fluid. There was no significant effect on ovulation rate or subsequent corpus luteum function as measured by plasma concentrations of progesterone. Infusion of ovine FSH (50 micrograms/h for 48 h) during the period of follicular fluid treatment prevented the delay in onset of oestrus and resulted in a substantial (2-10-fold) increase in ovulation rate. Corpus luteum function in terms of progesterone secretion was also enhanced. These results show that (1) intermittent suppression of FSH during the preovulatory period in the ewe does not affect subsequent ovulation rate or corpus luteum function and (2) the delay in the onset of oestrus induced by bovine follicular fluid can be prevented by exogenous FSH.     

The effect of epostane (Win32739), an inhibitor of 3β-hydroxy steroid dehydrogenase,on luteal function and gonadotrophin secretion in cows

Six cows were injected i.m. with either 4 × 125 mg or 4 × 250 mg of the 3β-hydroxy steroid dehydrogenase inhibitor epostane (Win32729) at 12-h intervals during the luteal phase of the oestrous cycle. Four more cows received 1 × 1 g epostane i.m. In all cows there was a transient decrease in plasma progesterone concentrations beginning within 8 h of the first injection, the decrease being more rapid and greater in the group receiving 1 × 1 g epostane. However, progesterone concentrations did not reach basal values and no preovulatory LH or FSH surges occurred. Progesterone concentrations invariably returned to pre-injection values within a few days and the length of the oestrous cycle was not affected. During the treatment period there were significant negative correlations between mean plasma LH and progesterone concentrations.     

Validation of the mechanisms proposed for the stimulatory and inhibitory effects of progesterone on gonadotropin secretion in the estrogen-primed rat: a possible role for adrenal steroids.

The stimulatory and inhibitory effects of progesterone on luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion were found to be dependent on the length of estrogen exposure in ovariectomized estrogen-primed rats. Progesterone suppressed LH and FSH secretion when administered 16 hours after a single injection of estradiol to ovariectomized rats. If the estradiol treatment was extended over 40 hours by two injections of estradiol 24 hours apart, progesterone administration led to a highly significant elevation of both serum LH and FSH levels 6 hours later. In addition to the direct stimulatory effect on LH and FSH release, progesterone, when injected 1 hour before, was able to antagonize the suppressive effect of a third injection of estradiol on LH and FSH release. In the immature ovariectomized estrogen-primed rat, 10 IU of ACTH brought about a release of progesterone and corticosterone 15 minutes later and LH and FSH 6 hours later. Progesterone, but not corticosterone, appeared to be responsible for the effect of ACTH on gonadotropin release. The synthetic corticosteroid triamcinolone acetonide brought about LH and FSH release in the afternoon, while cortisol, similar to corticosterone, was unable to do so. Nevertheless, triamcinolone acetonide and cortisol brought about increased secretion of FSH the following morning.     

Differential release of LH and FSH in cyclic mares in response to synthetic Gn-RH.

Mares at different stages of the oestrous cycle were given a single intravenous injection of 0.5 mg synthetic Gn-RH. The mean area of the induced LH peak was significantly less at mid-cycle (Day 10-11) than at any other time. The mean height of the LH peak above preinjection concentration was greater at late oestrus and early cycle (Day 5-6) than at mid-cycle and early oestrus. There were no significant different in mean FSH responses. The LH:FSH ratio for both height and area of induced peaks was significantly less at mid-cycle than at other times of injection. These results suggest that one releasing hormone could cause the release of both FSH and LH in the normal cyclic mare.     

Induction of ovulation in postpartum suckled beef cows: a review

Prolonged postpartum acyclicity in suckled beef cows reduces the calf crop, and causes economic loss to beef cattle producers. Once anterior pituitary LH stores have been replenished between Days 15 and 30 post partum in suckled beef cows, methods to initiate cyclicity include non-hormonal methods such as weaning of calves (either complete, temporary or partial), or exposure to bulls, and hormonal methods such as administration of GnRH (either single injection, intermittent injections, or continuous infusion), gonadotropins (eCG, FSH, hCG), and steroids (estrogens, anti-estrogens, and progestogens). Weaning is costly, reduces growth rate of weaned calves, and short cycles are common after weaning-induced ovulation. Exposure of cows to bulls is not practical and its effect is not predictable. Repeated injections of GnRH, or a single injection of hCG are not always effective; ovulation is always followed by a short cycle, and usually a return to acyclicity. Estrogens and anti-estrogens do not consistently shorten postpartum anestrus. Exogenous progestogens include intravaginal devices, such as controlled-internal drug release (CIDR) or progesterone-releasing intravaginal device (PRID), norgestomet implants, and the feed-additive melengestrol acetate (MGA). Administration of exogenous progestogens is more practical than, and offers more advantages over, other treatments to shorten postpartum acyclicity in suckled beef cows. Mimicking the short cycle after Week 3 post partum, by maintaining circulating progesterone at subluteal concentrations or circulating progestin at intermediate concentrations, extends the life-span and allows terminal maturation of the postpartum dominant follicle as in cyclic cows, by initiating endogenous GnRH and LH pulses. This is followed by an LH surge, ovulation and normal cycles. The benefit from using exogenous progestogens after Week 3 post partum in suckled beef cows is that ovulation is induced, cyclicity is initiated, the resulting CL has a normal life-span and function, and there is no need to change management, such as weaning of calves. We present a model for the induction of ovulation and initiation of cyclicity using exogenous progestogens after Week 3 post partum in suckled beef cows.     

Effect of exogenous FSH on estrus, ovulation and endogenous hormone release in dairy cows

Fifteen lactating Holstein cows were randomly allotted to receive either 0 mg (group 0), 32 mg (group 1) or 50 mg (group 2) porcine follicle stimulating hormone (FSH-P) injected in 10 fractions at 12 hr intervals beginning on day 9 of the estrous cycle. All cows received 25 mg prostaglandin (PG) on day 11. Jugular blood samples were collected from cows in all groups at 6 hr intervals beginning on day 7 and continuing through expression of estrus. Mean duration to occurrence of estrus and preovulatory LH surge after PG injection was reduced (P<.05) by injection of FSH-P. Mean number of ovulations increased (P<.05) progressively with increased dose of FSH-P. Mean peripheral progesterone declined more uniformly in FSH-P treated cows after PG and increased earlier (P<.05) after estrus in group 2 cows compared to group 0 and 1 cows. Mean plasma estradiol-17beta elevated (P<.05) after PG injection in both FSH-P-treated groups compared to group 0 cows. Both LH and FSH increased (P<.05) for 36 hr after initiation of FSH-P injection in groups 1 and 2, then declined until after PG injection. Peak LH and FSH occurred more uniformly following PG in treated cows. Results indicate that FSH-P increased endogenous gonadotropin release, estradiol-17beta, ovulation rate and reduced duration to estrus and preovulatory gonadotropin release after PG. Injection of 50 mg FSH-P increased plasma estradiol-17beta and ovulation rate compared to injection of 32 mg FSH-P.     

Effects of a systematic application of human chorionic gonadotropin, gonadotropin-releasing hormone analog and bovine anterior pituitary gonadotropin in cows with cystic ovarian disease

Fourty-one cows with ovarian follicular cysts (cysts) diagnosed by rectal palpation and observation of estrus behavior were injected intramuscularly (i.m.) with 20,000 IU of human chorionic gonadotropin (HCG). Ten days after this treatment all the cows were examined per rectum for changes in the ovaries with special regard to luteinization of cysts. Cows not responding to HCG were administered 500(10)mug of an analog of gonadotropin-releasing hormone (Gn-RH), Des-Gly-LH-RH-ethylamide, i.m.. If the cysts remained unchanged 10 days subsequent to the second treatment, a third treatment, 400 KE of bovine anterior pituitary gonadotropin (APG), was given i.m. HCG was clinically effective in only 8 cows (20 %). All of the 8 cows conceived. Of 33 cows not responding to HCG, 18 cows (55 %) responded to Gn-RH analog and 12 cows (36 %) conceived. Eleven (73 %) of 15 cows failing to respond to Gn-RH analog were successfully treated with APG and 6 cows (40 %) conceived. In 37 cases, treatment effects were also evaluated by determining serum levels of progesterone prior to and 10 days subsequent to each treatment. When effects of treatment were judged by 1.0 ng/ml or more increase of serum progesterone levels 10 days after treatment, HCG was effective in 13 of 37 cows (35 %), retreatment with Gn-RH analog was successful in 8 of 16 cows (50 %) and APG was ineffective in 3 cows not responding to both HCG and Gn-RH analog. It may be concluded that the therapeutic effect of HCG is disappointing and about half of the cases not responding to HCG were successfully treated with Gn-RH analog. If the cows did not respond to both HCG and Gn-RH analog, they may not respond to APG either.     

Serum progesterone concentration in pyometritic and normal postpartum dairy cows

Two studies were conducted to evaluate the concentration of serum progesterone in pyometritic cows and relate it to palpation of ovarian structures per rectum . In Trial 1, serum samples from 34 pyometritic cows were assayed for progesterone. Mean serum progesterone concentrations were 6.8 +/- 0.7 ng/ml. In Trial 2, each of 54 pyometritic cows was paired with a control cow on the basis of days post partum (18-50 days). Mean concentration of progesterone was 9.7 +/- 1.0 ng/ml for the pyometritic cows and 5.7 +/- 0.8 ng/ml in control cows (P<0.005). Progesterone concentration was greater (P<0.005) in both groups of cows with palpable corpora lutea (CLs). Ninety-six percent of the pyometritic cows had palpable CLs compared to 57% of the control group. Comparing serum progesterone only in cows with a palpable CL, the mean concentration was still greater (P<0.005) in the pyometritic group (10.6 +/- 1.0 ng/ml) than the control group (6.6 +/- 1.0 ng/ml). Compatability of rectal palpation findings and concentrations of serum progesterone were 92.6% for the pyometritic group and 72.2% for the control group. Progesterone concentration increased (P<0.05) by increased days post partum in Trial 2 (n=54) but not in Trial 1 (n=23). In both Trials 1 and 2, uterine size due to pyometra increased (P<0.05) with increased days post partum. No other associations were found.