Thyroid stimulating hormone and prolactin secretion after thyrotropin releasing hormone administration to mares: Dose response during anestrus in winter and during estrus in summer

The response of thyroid stimulating hormone (TSH) and prolactin (PRL) concentrations to administration of thyrotropin releasing hormone (TRH) was determined in light-horse mares during the anestrous season (winter) and during estrus (standing heat) in the summer. Within each season, mares (4/group) were treated with either saline (controls) or one of four doses of TRH (80, 400, 2,000 or 10,000 ug) intravenously. Samples of blood were drawn at −15, −.5, 15, 30, 45, 60, 90, 120, 180 and 240 min relative to TRH injection. Concentrations of TSH and PRL in pre-TRH samples were greater (P<.05) in anestrous mares during winter than in estrous mares during summer. Concentrations of TSH increased (P<.05) within 30 min after administration of TRH and remained elevated during the 4-hr sampling period. The maximal net change in TSH concentrations and the area under the response curve were greatest for 2,000 ug of TRH; 80 ug did not produce a significant TSH response. There was no interaction (P >.10) between reproductive state and TRH dose for TSH concentrations. Concentrations of PRL were not significantly affected by any TRH dose during either season. It appears that mares differ from many mammalian species in that they do not respond to an injection of TRH with increases in both TSH and PRL.     

Serum concentrations of prolactin, thyroxine and triiodothyronine relative to season and the estrous cycle in the mare

Studies were conducted to characterize circulating concentrations of prolactin, thyroxine (T4) and triiodothyronine (T3) relative to season and the estrous cycle, and to evaluate the ability of thyrotropin-releasing hormone (TRH) to induce prolactin secretion in the horse mare. The increase in serum prolactin following the iv injection of 0, 5, 50 and 500 micrograms TRH was dose-related, while all three doses of TRH induced a significant and comparable increase in serum T4. Seasonal variations in serum prolactin were directly correlated (P less than .001) with changes in photoperiod (r = .80) and temperature (r = .61). By comparison, concentrations of T3 were inversely related to temperature (r = -.49; P less than .001), while changes in T4 were less closely associated with photoperiod (r = -.29; P = .04). There were no significant fluctuations in prolactin, T3 or T4 at any stage of the estrous cycle. These results demonstrate that concentrations of prolactin in the mare are highest during the summer and lowest during the winter, but it is unknown whether this annual rhythm is controlled by photoperiod, temperature and(or) other environmental factors.     

Thyroxine concentrations are elevated in mares which continue to exhibit estrous cycles during the nonbreeding season

In two successive years, the pattern of secretion of thyroxine was compared in seasonally anestrous mares and a subpopulation of mares that exhibited estrous cycles during the nonbreeding season. Blood samples were collected two or three times per week between October and April from a total of 15 mares that exhibited estrous cycles during the nonbreeding season and 21 anestrous mares. Monthly mean concentrations of thyroxine during the period from October through April were greater (p<0.05) in cycling versus anestrous mares. However, it remains to be determined whether continuation of estrous cycles during the nonbreeding season and elevated concentrations of thyroxine are causally related.     

Concentrations of prolactin, luteinizing hormone and follicle stimulating hormone in pituitary and serum of horses: effect of sex, season and reproductive state

Pituitary and serum from 86 male or female horses of various reproductive states were collected in the normal breeding season (summer) and in the nonbreeding season (winter) at a commercial slaughterhouse. Concentrations of prolactin (PRL), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay. Concentrations of pregnant mare serum gonadotropin and reproductive steroids in serum and gross appearance of the reproductive tract and gonads were used to catagorize reproductive state. Concentrations of PRL were higher (P less than .01) in summer than in winter in pituitary and serum of mares, stallions and geldings. In summer, mares had higher (P less than .01) concentrations of PRL in serum than stallions. In mares, concentrations of LH in pituitary were higher (P less than .05) in summer than in winter. Concentrations of LH in serum were higher (P less than .01) in summer than in winter in mares and geldings, higher (P less than .01) in mares than in stallions in summer, higher (P less than .01) in geldings than in stallions in summer and higher (P less than .01) in mares with low serum progesterone (P) concentrations than in mares with high P concentrations in summer. Concentrations of FSH in pituitary and serum did not differ between summer and winter for any type of horse.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estradiol interactions with dopamine antagonists in mares: Prolactin secretion and reproductive traits

Two experiments studied the effects of pretreatment with estradiol benzoate before treatment with a dopamine antagonist on prolactin secretion and reproductive traits in mares during (1) the seasonal anovulatory period and (2) the normal breeding season. Experiment 1 was performed in winter with 17 mares selected for low follicular activity. Nine mares received estradiol benzoate injections every other day for a total of 10 injections; 8 mares received similar injections of vehicle. Ten days after onset of injections, all mares were placed on daily injections of sulpiride (250 mg) for 35 days or until ovulation. Plasma prolactin concentrations were higher (P < .001) in mares receiving estradiol than in controls for all assessments from days 12 through 36. Plasma luteinizing hormone (LH) concentrations were also increased (P < .05) by estradiol treatment from days 14 to 23. Mean day of first ovulation was 73.6 for control mares and 29.0 for estradiol-treated mares (P = .016). Estradiol treatment greatly enhanced prolactin secretion in response to sulpiride and increased LH secretion in seasonally anovulatory mares, which together hastened the date of first ovulation by an average of 45 days. Experiment 2 was designed to assess the efficacy of a long-acting, single-injection microparticle preparation of another dopamine antagonist, domperidone, for increasing prolactin secretion in cyclic mares in the summer. The experimental design and procedures used in experiment 1 were repeated, except that a single 3-g domperidone-microparticle injection was administered on day 11 rather than 45 days of sulpiride injections. Day 0 was the first day of estrus for each mare. Prolactin concentrations were higher (P < .05) in mares receiving estradiol than in control mares from days 12 through 25 and after a thyrotropin-releasing hormone injection on d 21. Estrous cycle traits (time to ovulation and time of luteal regression) were not affected (P > .1) by treatment. Estradiol enhanced the prolactin response to a single injection of 3 g domperidone in cyclic mares in the summer in a manner similar to the estradiol enhancement of prolactin secretion in response to daily sulpiride injections in anovulatory mares in winter. Thus, the single injection of domperidone could possibly replace the daily sulpiride injections used in experiment 1 to induce ovulation in seasonally anovulatory mares; this needs to be tested in future experiments.     

Growth hormone in mares and stallions: pulsatile secretion, response to growth hormone-releasing hormone, and effects of exercise, sexual stimulation, and pharmacological agents.

Short-term patterns of growth hormone (GH) secretion and factors affecting it were studied in mares and stallions. In Exp. 1, hourly blood samples were collected from three mares and three stallions in summer and winter. Although GH concentrations varied in a pulsatile manner in all horses, there was no effect of sex or season (P greater than .1) on plasma GH concentrations and no indication of a diurnal pattern of GH secretion. In Exp. 2, 10-min blood samples were drawn for 8 h from 12 mares; after 6 h, porcine GH-releasing hormone (GHRH) was administered i.v. at 0, 45, 90, or 180 micrograms/mare (three mares per dose). Pulsatile secretion of GH occurred in all mares and averaged 2.4 +/- .3 peaks/6 h; amplitudes were variable and ranged from 2.6 to 74.4 ng/mL. Eight of nine mares responded within 20 min to GHRH injection, but there was no difference (P greater than .1) among the three doses tested. In Exp. 3, plasma GH concentrations in stallions increased (P less than .05) 8- to 10-fold after 5 min of acute physical exercise or exposure to an estrual mare. Restraint via a twitch (5 min) and epinephrine administration (3 mg i.v.) also increased (P less than .05) plasma GH concentrations by approximately fourfold. In Exp. 4 and 5, administration of either .4, 2, or 10 mg of thyrotropin-releasing hormone (TRH) or 100 or 500 mg of sulpiride (a dopamine receptor antagonist) increased (P less than .01) plasma prolactin concentrations but had no effect (P greater than .1) on GH concentrations during the same period of time.(ABSTRACT TRUNCATED AT 250 WORDS)     

Secretion of luteinizing hormone and follicle stimulating hormone in intact and ovariectomized mares in summer and winter

Sequential samples of blood were drawn via jugular catheters every 15 min for 24 h from four mares in each of five reproductive states: intact anestrous mares in winter, intact diestrous mares in summer, intact estrous mares in summer, ovariectomized mares in winter and ovariectomized mares in summer. Estrous mares were sampled on d 4 or 5 of estrus and diestrous mares on d 10 or 11 of diestrus. Each sample of plasma was assessed for concentrations of luteinizing hormone (LH) and follicle stimulating hormone (FSH) in two independent radioimmunoassays. A computer program was developed that determined peak hormone concentrations based on assay sensitivity, assay variability and repeatability of peaks in both independent assays. Peaks in LH and FSH were observed for mares in all five reproductive states, except for FSH concentrations in estrous mares. High frequency peaks of short duration were observed only in ovariectomized mares. Low frequency peaks of relatively long duration were observed in both intact and ovariectomized mares in both seasons. With the exception of estrous mares, there was variation among mares in the patterns of LH and(or) FSH within any one group; all estrous mares exhibited high, variable LH concentrations and low, constant FSH concentrations. In general, peaks in both gonadotropins occurred simultaneously. Ovariectomized mares exhibited more (P less than .05) peaks/24 h than intact mares for both gonadotropins. Ovariectomized mares also exhibited more (P less than .05) FSH peaks/24 h in summer than in winter.     

Evidence for a seasonal variation in the ability of exogenous melatonin to suppress prolactin secretion in the mare

In seasonally breeding species photoperiodic information is thought to be conveyed to the reproductive and prolactin axis via changes in circulating concentrations of melatonin. For some species, a constant melatonin stimulus is perceived as a short day, whereas in others no photoperiodic information is provided. In the mare, a preliminary study demonstrated that constant administration of melatonin did not modify prolactin secretion, suggesting that this treatment regimen failed to provide photoperiodic information. To further investigate this proposal and to investigate an alternative explanation, namely a seasonal variation in response to melatonin, 4 experiments were performed. In experiments 1–3, the effects of constant administration of melatonin on prolactin secretion were investigated. In each study the time of treatment initiation varied beginning before the summer solstice, (May 9; Exp. 1), at the autumnal equinox (Sept. 21; Exp. 2) or the winter solstice (Dec. 21; Exp. 3). In Experiment 4, melatonin was administered as a timed daily injection (5 PM) for 6 months, beginning at the summer solstice (June 21). Constantly elevated physiological concentrations of melatonin (expts. 1–3) and an extended nighttime elevation of melatonin (exp. 4) suppressed prolactin concentrations only during the spring and early summer months (April–August). At other times during the year prolactin concentrations were similar to untreated mares. In the presence of a continuous melatonin implant the circannual rhythm of prolactin secretion was not disturbed. The results suggest that the prolactin axis of the mare is sensitive to an inhibitory melatonin signal during a restricted period of time and that at other times is refractory to this signal.     

Plasma concentrations of cortisol, prolactin, luteinizing hormone, and follicle-stimulating hormone in stallions after physical exercise and injection of secretagogue before and after sulpiride treatment in winter

Ten lighthorse stallions were used to determine 1) whether prolactin (PRL) and cortisol responses previously observed after acute exercise in summer would occur in winter when PRL secretion is normally low, 2) whether subsequent treatment with a dopamine receptor antagonist, sulpiride, for 14 d would increase PRL secretion and response to thyrotropin-releasing hormone (TRH) and exercise, and 3) whether secretion of LH, FSH, and cortisol would be affected by sulpiride treatment. On January 11, blood samples were drawn from all stallions before and after a 5-min period of strenuous running. On January 12, blood samples were drawn before and after an i.v. injection of GnRH plus TRH. From January 13 through 26, five stallions were injected s.c. daily with 500 mg of sulpiride; the remaining five stallions received vehicle. The exercise and secretagogue regimens were repeated on January 27 and 28, respectively. Before sulpiride injection, concentrations of both cortisol and PRL increased (P less than .05) 40 to 80% in response to exercise; concentrations of LH and FSH also increased (P less than .05) approximately 5 to 10%. Sulpiride treatment resulted in (P less than .05) a six- to eightfold increase in daily PRL secretion. The PRL response to TRH increased (P less than .05) fourfold in stallions treated with sulpiride but was unchanged in control stallions. Sulpiride treatment did not affect (P greater than .05) the LH or FSH response to exogenous GnRH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of physiologic and pharmacologic agents on serum prolactin concentrations in the nonpregnant mare

Four studies were conducted to evaluate the effects of several physiologic and pharmacologic agents on serum prolactin concentrations in the nonpregnant mare. An increase in prolactin measured in response to administration of thyrotropin-releasing hormone (TRH; 50 micrograms, iv) was found not to vary (P = .20) in mares in estrus compared with mares in diestrus (5 to 10 d post-ovulation). Administration in the dopamine receptor blocker, metoclopramide (25 or 100 mg, im), rapidly increased serum prolactin, and the response was dependent on dose administered (total prolactin measured for 420 min was 3,362.7 +/- 182.1 ng for 25 mg, and 4,485.7 +/- 212.6 ng for 100 mg administered im; P less than .05), but not on route of injection (3,026.3 +/- 492.3 ng prolactin with 25 mg, iv; P less than .05). Similarly, sulpiride, a D-2 dopamine receptor blocker, induced an increase in serum prolactin, which appeared to be maximal at a dose of 25 mg (6,556.3 +/- 636.9 ng prolactin/420 min compared with 6,594.5 +/- 169.3 ng prolactin/420 min with 100 mg sulpiride; P less than .10). Finally, bromocriptine, a dopamine agonist, decreased serum prolactin compared with vehicle-injected controls, but the inhibitory effect was found only when basal levels of serum prolactin were highest (in May). These data suggest that mechanisms controlling prolactin secretion in the mare are similar to those described in other mammalian species, and that the seasonal decline in serum prolactin is not the result of increased sensitivity to the proposed prolactin-inhibiting factor, dopamine.     

Thyrotropin releasing hormone interactions with growth hormone secretion in horses

Light horse mares, stallions, and geldings were used to 1) extend our observations on the thyrotropin releasing hormone (TRH) inhibition of GH secretion in response to physiologic stimuli and 2) test the hypothesis that stimulation of endogenous TRH would decrease the normal rate of GH secretion. In Exp. 1 and 2, pretreatment of mares with TRH (10 microg/kg BW) decreased (P < 0.001) the GH response to exercise and aspartate infusion. Time analysis in Exp. 3 indicated that the TRH inhibition lasted at least 60 min but was absent by 120 min. Administration of a single injection of TRH to stallions in Exp. 4 increased (P < 0.001) prolactin concentrations as expected but had no effect (P > 0.10) on GH concentrations. Similarly, 11 hourly injections of TRH administered to geldings in Exp. 5 did not alter (P > 0.10) GH concentrations either during the injections or for the next 14 h. In Exp. 5, it was noted that the prolactin and thyroid-stimulating hormone responses to TRH were great (P < 0.001) for the first injection, but subsequent injections had little to no stimulatory effect. Thus, Exp. 6 was designed to determine whether the inhibitory effect of TRH also waned after multiple injections. Geldings pretreated with five hourly injections of TRH had an exercise-induced GH response identical to that of control geldings, indicating that the inhibitory effect was absent after five TRH injections. Retrospective analysis of pooled, selected data from Exp. 4, 5, and 6 indicated that endogenous GH concentrations were in fact lower (P < 0.01) from 45 to 75 min after TRH injection but not thereafter. In Exp. 7, 6-n-propyl-2-thiouracil was fed to stallions to reduce thyroid activity and hence thyroid hormone feedback, potentially increasing endogenous TRH secretion. Treated stallions had decreased (P < 0.01) concentrations of thyroxine and elevated (P < 0.01) concentrations of thyroid-stimulating hormone by d 52 of feeding, but plasma concentrations of GH and prolactin were unaffected (P > 0.10). In contrast, the GH response to aspartate and the prolactin response to sulpiride were greater (P < 0.05) in treated stallions than in controls. In summary, TRH inhibited exercise- and aspartate-induced GH secretion. The duration of the inhibition was at least 1 h but less than 2 h, and it waned with multiple injections. There is likely a TRH inhibition of endogenous GH episodes as well. Reduced thyroid feedback on the hypothalamic-pituitary axis did not alter basal GH and prolactin secretion.     

Prolactin and Gonadotropin Responses in Geldings to Injections of Estradiol Benzoate in Oil, Estradiol Benzoate in Biodegradable Microspheres, and Estradiol Cypionate

We previously reported success in inducing early ovulation in seasonally anovulatory mares with a combination of estradiol pretreatment followed by daily administration of a dopamine antagonist (sulpiride). Although every-other-day injections of estradiol benzoate (EB) were effective in that experiment, practical application of this technology would require simplification of the treatment regimen. The current experiment was designed to compare, in a gelding model, the biologic responses of two alternative, one-injection regimens for estradiol delivery to the established EB treatment used previously. Fifteen long-term geldings were sampled via jugular venipuncture from November 5 to 7, 2006, and were then administered intramuscular injections of vegetable oil (n = 4); EB, 11 mg in oil (n = 4; controls); EB in biodegradable microspheres (300 mg; n = 3); or estradiol cypionate, 100 mg in oil (n = 4). Injections of EB in oil were repeated every other day for a total of 10 injections, as was done in our previous experiment. Jugular blood samples were drawn from all geldings at 3, 6, 12, 24, 36, and 48 hours relative to injections, and then on the mornings of days 3, 4, 6, 8, 10 to 18, 22, 26, and 30. On days 10 through 13, all geldings received subcutaneous injections of 125 mg sulpiride, a dopamine receptor antagonist, to stimulate prolactin secretion. On day 12, each gelding received an intravenous injection of 30 μg gonadotropin-releasing hormone (GnRH) analog and 3 mg thyrotropin-releasing hormone (TRH); frequent blood samples were drawn to characterize the luteinizing hormone (LH), follicle-stimulating hormone (FSH), and prolactin responses. Relative to geldings receiving oil, all geldings receiving estradiol injections had a rise (P < .05) in estradiol concentrations lasting at least 12 days. Daily LH concentrations increased (P < 0.01) in all treated groups, but the response was delayed approximately 14 days in the geldings receiving EB in microspheres. Daily FSH concentrations decreased (P < .01) in all treated groups, with the greatest response in the geldings receiving EB in microspheres. Prolactin in daily samples increased (P < .01) similarly in all estradiol-treated groups after injection of sulpiride. The LH response to GnRH analog was greatest (P < .05) in geldings receiving EB in oil and estradiol cypionate; the FSH response was not altered by treatment. The prolactin response to TRH was greater (P < .01) in estradiol-treated geldings relative to controls, but did not differ among groups. Compared with the responses to every-other-day EB injections in oil, as we used previously, a single injection of 100 mg estradiol cypionate gave the most similar and consistent responses. Because of these similar responses in this gelding model, it is likely that a single injection of 100 mg estradiol cypionate can be used in lieu of every-other-day injections of EB in oil in the treatment regimen we reported previously for stimulating ovarian activity in seasonally anovulatory mares.     

Estradiol Effects on Secretagogue-Induced Prolactin Release: Disparity in Responses to Sulpiride,Exercise, Epinephrine,Prostaglandin-F2α, and Thyrotropin-Releasing Hormone

Three experiments were conducted (1) to assess the effects of estradiol pretreatment on the prolactin response to various secretagogues, and (2) to determine whether elevated plasma thyroxine concentrations altered the prolactin responses to those secretagogues. Geldings were available and were used because their prolactin and luteinizing hormone responses to estradiol and dopamine antagonists are known to be similar to those in seasonally anovulatory mares. In the first experiment, performed in summer, estradiol cypionate (ECP; 100 mg) treatment of geldings increased (P = .07) plasma prolactin concentrations before the onset of exercise, and repeated exercise bouts stimulated (P < .001) plasma prolactin concentrations after each bout; there was no interaction with estradiol pretreatment. Epinephrine injection (5 μg/kg of body weight) did not alter prolactin concentrations. Prostaglandin-F_2α administration (10 mg Lutalyse) stimulated (P < .001) prolactin concentrations, but there was no interaction with ECP pretreatment. Sulpiride administration (0.1 mg/kg of body weight) stimulated (P < .001) prolactin concentrations, and there was a greater (P = .038) response in ECP-treated geldings relative to controls. In the second experiment, performed in winter, ECP (50 mg) pretreatment of geldings before 21 days of daily thyrotropin-releasing hormone (TRH; 1.5 mg) injections did not alter prolactin secretion (P > .1); TRH stimulated prolactin secretion only after the very first injection. In the third experiment (performed in July), pretreatment of geldings with 50 mg of thyroxine in biodegradable particles (day 0) raised (P < .001) plasma thyroxine concentrations in plasma for the duration of the experiment, but had no effect on the prolactin responses to two exercise bouts on day 5, to an injection of prostaglandin-F_2α on day 9, or to an injection of sulpiride on day 13. The previously reported stimulation of plasma prolactin concentrations by estradiol pretreatment and subsequent sulpiride administration in mares, as evidenced herein in geldings, does not occur when prolactin is stimulated by exercise, prostaglandin-F_2α, or TRH. The practical impact of these data is that stimulation of prolactin concentrations after ECP treatment in winter, in an effort to stimulate ovarian activity in seasonally anovulatory mares, is likely limited to dopamine antagonists. Results of the third experiment indicate that TRH is not likely the mediator in the prolactin response to exercise or prostaglandin-F_2α injection.     

The relationship between body condition,leptin, and reproductive and hormonal characteristics of mares during the seasonal anovulatory period

An experiment was conducted to determine the effects of high vs low body condition scores (BCS) produced by restricted feeding on reproductive characteristics, hormonal secretion, and leptin concentrations in mares during the autumnal transition and winter anovulatory period. Mares with BCS of 6.5 to 8.0 were maintained on pasture and/or grass hay, and starting in September, were full fed or restricted to produce BCS of 7.5 to 8.5 (high) or 3.0 to 3.5 (low) by December. All but one mare with high BCS continued to ovulate or have follicular activity during the winter, whereas mares with low BCS went reproductively quiescent. Plasma leptin concentrations varied widely before the onset of restriction, even though all mares were in good body condition. During the experiment, leptin concentrations gradually decreased (P < 0.0001) over time in both groups, but were higher (P < 0.009) in mares with high vs low BCS after 6 wk of restriction, regardless of initial concentration. No differences (P > 0.1) between groups were detected for plasma concentrations of LH, FSH, TSH, GH, glucose, or insulin in samples collected weekly; in contrast, plasma prolactin concentrations were higher (P < 0.02) in mares with high BCS, but also decreased over time (P < 0.008). Plasma IGF-I concentrations tended (P = 0.1) to be greater in mares with high vs low BCS. The prolactin response to sulpiride injection on January 7 did not differ (P > 0.1) between groups. During 12 h of frequent blood sampling on January 12, LH concentrations were higher (P < 0.0001), whereas GH concentrations (P < 0.0001) and response to secretagogue (EP51389; P < 0.03) were lower in mares with high BCS. On January 19, the LH response to GnRH was higher (P < 0.02) in mares with high BCS; the prolactin response to TRH also was higher (P < 0.01) in mares with high BCS. In conclusion, nutrient restriction resulting in low BCS in mares resulted in a profound seasonal anovulatory period that was accompanied by lower leptin, IGF-I, and prolactin concentrations. All but one mare with high BCS continued to cycle throughout the winter or had significant follicular activity on the ovaries. Although leptin concentrations on average are very low in mares with low BCS and higher in well-fed mares, there is a wide variation in concentrations among well-fed mares, indicating that some other factor(s) may determine leptin concentrations under conditions of high BCS.     

Effects of age, season and active immunization against estrogen on serum prolactin concentrations in stallions

Prolactin concentrations in stallion serum were measured by radioimmunoassay based on antiserum generated against equine prolactin and radioiodinated canine prolactin. Prolactin concentrations in serum collected from 152 stallions at a slaughterhouse were higher (P less than .01) in summer than in winter (5.7 +/- .15 vs 2.0 +/- .17 ng/ml). Moreover, there was an effect of age (P less than .02) in the analysis of variance; there was no interaction between age and season. In general, prolactin concentrations increased with age up to 3 to 5 years. Samples of serum collected from five control and five estrogen-immunized stallions from 6 to 26 months of age also were assessed for prolactin concentration. Active immunization against estrogen had no effect (P greater than .10) on prolactin concentrations. In contrast, whether analyzed by age or by month of the year, prolactin concentrations varied with time (P less than .01) in these stallions. Averaged over both groups, concentrations were high during summer and low during winter and were also gradually increasing with time as indicated by a linear trend (P less than .01) in the data. It is concluded that serum prolactin concentrations in stallions 1) are seasonal, as in many other seasonally breeding species, 2) in general are highest in older stallions and lowest in young stallions and 3) are not affected by active immunization against estrogens.     

Responses of seasonally anovulatory mares to daily administration of thyrotropin-releasing hormone and(or) gonadotropin-releasing hormone analog

Seventeen seasonally anovulatory light horse mares were treated daily, starting January 5 (d 1), for 28 d with GnRH analog (GnRH-A; 50 ng/kg BW) and(or) thyrotropin-releasing hormone (TRH; 5 microg/kg BW) in a 2 x 2 factorial arrangement of treatments to test the hypothesis that combined treatment may stimulate follicular growth and development. Ovaries were examined via ultrasonography and jugular blood samples were collected every 3 d. Frequent blood samples were collected after treatment injections on d 1, 2, 4, 7, 11, 16, and 22; on d 29, all mares received an i.v. mixture of GnRH, TRH, sulpiride, and EP51389 (a growth hormone secretagogue) to assess pituitary responsiveness. No consistent effects (P > 0.1) of treatment were observed for plasma LH, FSH, prolactin, or thyroxine concentrations in samples collected every 3 d. The only effect on ovarian follicle numbers was a reduction in number of follicles 11 to 19 mm in diameter due to TRH treatment (P = 0.029). No mare ovulated during treatment. On the days of frequent sampling, mean LH (P = 0.0001) and FSH (P = 0.001) concentrations were higher in mares receiving GnRH-A and tended to increase from d 1 through 7. In contrast, mean prolactin (P = 0.001) and thyroid-stimulating hormone (P = 0.0001) concentrations were high in mares receiving TRH on d 1 but rapidly decreased thereafter. When mares were administered the secretagogue mixture on d 29, the LH response was greater (P = 0.0002) in mares that had previously received GnRH-A but the FSH response was not affected (P > 0.1); the prolactin response was greater (P = 0.014) and the TSH response was smaller (P = 0.0005) in mares that had previously received TRH. Surprisingly, an immediate growth hormone response to EP51389 was absent in all mares. In conclusion, daily GnRH-A treatment stimulated plasma LH and FSH concentrations immediately after injection; although no long-term elevation in preinjection concentrations was achieved, the responses gradually increased over time, indicating a stimulation of gonadotropin production and storage. Daily treatment with TRH stimulated plasma TSH and prolactin concentrations, but the response diminished rapidly and was minimal within a few days, indicating a depletion of pituitary stores and little or no stimulation of production. There was no beneficial effect of adding TRH treatment to the daily GnRH-A regimen.     

Dopaminergic-like activity in toxic fescue alters prolactin but not growth hormone or thyroid stimulating hormone in ewes

Studies were conducted to determine the specificity and cause of altered pituitary hormone secretion when ewes ingest endophyte-infected (Acremonium coenophialum) GI-307 tall fescue (toxic fescue). Plasma concentrations of prolactin (PRL) but not growth hormone (GH) or thyroid stimulating hormone (TSH) in ewes grazing toxic fescue were significantly lower (P < .01) than concentrations measured in ewes grazing orchardgrass (OG). Comparing hormone secretory responses of ewes grazing each grasstype, ewes on toxic fescue released less PRL following thyrotropin releasing hormone (TRH) challenge than ewes on OG. TSH responses to TRH were not affected by grasstype. At this dose of TRH, GH secretion was not significantly affected in either group of ewes. In a separate study, dopamine hydrochloride (DA) was infused into control ewes to define the effect of a pure dopamine agonist on basal and TRH-stimulated secretion of PRL, GH and TSH. DA depressed both basal and TRH-stimulated secretion of PRL without affecting the basal concentrations or responses of GH or TSH. Based on the assumption that the active agent in toxic fescue responsible for the observed hypoprolactinemia was a dopaminergic agonist, haloperidol (HAL), a DA receptor blocking drug, was administered to ewes grazing toxic fescue or OG. HAL evoked significant PRL secretion unaccompanied by any GH or TSH effect in both toxic fescue and OG ewes. Administration of HAL resulted in a gradual increase over 4 hr in PRL in toxic fescue ewes and prolonged the duration of the PRL response to TRH. No differences in circulating plasma concentrations of DA, epinephrine or norepinephrine were measured in ewes on troxic fescue or OG.

Alterations in pituitary hormone secretion due to toxic factors in fescue were confined to PRL. Hormone secretory responses to TRH and HAL suggest that the effects on PRL are mediated through dopamine-like activity in toxic fescue.     

Exogenous opioids increase plasma prolactin in Holstein calves primarily via a dopaminergic mechanism.

A study was conducted to determine whether exogenous opioids increase prolactin (PRL) secretion in Holstein heifer calves via a dopaminergic mechanism. Twenty-four Holstein heifer calves ranging in age from 5 to 7 mo were assigned to one of four treatment groups (six/treatment): 1) injection of saline (SAL); 2) injection of a synthetic enkephalin (D-Ala2, N-Me-Phe4, Met(O)5-ol enkephalin; DAMME); 3) injection of DAMME after pretreatment with the long-acting dopamine agonist 2-bromo-alpha-ergocryptine; or 4) injection of thyrotropin-releasing hormone (TRH) after pretreatment with 2-bromo-alpha-ergocryptine. Calves were equipped with indwelling jugular cannulas on d 1, and baseline plasma PRL concentrations were established. Animals receiving 2-bromo-alpha-ergocryptine were injected s.c. 3 h after the last baseline sample was drawn on d 1. On d 2, calves assigned to receive SAL, DAMME, or TRH were injected 2 h after the start of sampling, and sampling was continued for an additional 4.5 h. Basal plasma PRL was lower (P less than .01) on d 2 in calves injected with 2-bromo-alpha-ergocryptine than baseline levels on d 1. Plasma PRL was higher (P less than .01) in calves not pretreated with 2-bromo-alpha-ergocryptine after DAMME injection on d 2 but was not different after DAMME injection in calves pretreated with 2-bromo-alpha-ergocryptine. In contrast, plasma PRL increased (P less than .01) after TRH injection on d 2 in calves pretreated with 2-bromo-alpha-ergocryptine.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estrus, ovulation, and serum progesterone, estradiol, and LH concentrations in mares after an increased photoperiod during winter

On December 11, 1974, 15 seasonally anestrous mares were assigned at random to 1 of 3 experimental groups: outdoor-control, indoor-control, or indoor light-treated (a 16-hour photo-period). This experiment was terminated on April 21, 1975. The five mares in the indoor light-treated group ovulated 59.0+/-6.9 days later, which was 74 days earlier (P less than 0.01) than 2 of the 5 outdoor-controls (the other 3 ovulated after April 21 during a subsequent experiment) and 50 days earlier (P less than 0.05) than the indoor-controls. Durations of the 1st estrus for the 3 groups of mares were 13.3+/-3.6, 8.4+/-2.0, and 6.0+/-1.0 days for the indoor light-treated, indoor-control, and outdoor-control groups, respectively. The indoor light-treated mares averaged 4.2 estrous cycles before April 21, the indoor-control mares averaged 1.4 estrous cycles, and 2 of 5 outdoor-control mares ovulated 1 time during the experiment. The peripheral blood luteinizing hormone (LH), estradiol, and progesterone concentrations were minimal during winter anestrous. The hormone changes normally associated with estrous cycle activity in mares--maximal estradiol and luteinizing hormone concentrations near ovulation and maximal progesterone concentration during diestrus--were observed in all mares beginning at the 1st estrus. Hair loss was observed earlier in the light-treated mares, than in either of the other groups. In conclusion, a 16-hour photo-period initiated in early December for anestrous brood mares caused endocrinologically normal estrous cycles to begin within 2 months. This may allow breeding and foaling considerably earlier than normally expected.     

Seasonal and lactational effects on response to thyrotropin releasing hormone injection in Holstein cows

Fifty-four lactating Holstein cows were used to define relationships between stage of lactation and environmental measures on thyrotropin releasing hormone (TRH)-induced secretion of triiodothyronine (T₃, thyroxine (T₄) and prolactin. Mean black globe temperatures were -.3, 20.6, and 25.5C for winter, spring, and summer months, respectively. The circulating hormone responses of T₃, T₄ and prolactin were significantly affected by season, sampling time and there was a season by sampling time interaction. T₄ response was affected by stage of lactation, while prolactin responses differed by stage of lactation across the three seasons. Both T₃ and T₄ responses were lowest in winter and may indicate greater utilization rates to meet metabolic demands. T₃ responses peaked near 3 hr post TRH, while T₄ increased to the end of the 7 hr sampling period. Prolactin responses to TRH increased from winter through summer and were responsive to changes in ambient temperature and photoperiod. T₄ response increased with advancing stage of lactation and could be due to changes in metabolic activity and clearance. Spring TRH-induced prolactin response decreased with advancing lactation whereas, the response increased as lactation progressed during the summer. Monthly milk yield was affected by stage of lactation and lactation number. Early lactation cows produced most milk and milk yield peaked in third lactation. Differential responses to TRH may reflect alterations in milk yield during different stages of lactation.