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Sympathoadrenal stimulation may perturb results of endocrine tests performed on fractious horses. Sedation may be beneficial; however, perturbation of results may preclude useful information. Four experiments were designed to 1) determine the effects of epinephrine on insulin response to glucose (IR2G), 2) assess the effects of detomidine (DET), alone or combined with butorphanol (DET/BUT), on IR2G and glucose response to insulin (GR2I), and 3) assess the effects of BUT alone on IR2G. In Experiment 1, mares were administered saline or epinephrine (5 μg/kg BW) immediately before infusion of glucose (100 mg/kg BW). Glucose stimulated (P < .05) insulin release in controls at 5 minutes that persisted through 30 minutes; insulin was suppressed (P < .05) by epinephrine from 5 to 15 minutes, rising gradually through 30 minutes. Experiments 2 (IR2G) and 3 (GR2I) were conducted as triplicated 3 × 3 Latin squares with the following treatments: saline (SAL), DET, and DET/BUT (all administered at .01 mg/kg BW). Glucose stimulated (P < .05) insulin release that persisted through 30 minutes in SAL mares; DET and DET/BUT severely suppressed (P < .0001) the IR2G. Sedation did not affect resting glucose and had inconsistent effects on the GR2I when mares were treated with 50 mIU/kg BW recombinant human insulin. Butorphanol had no effect on IR2G. In conclusion, adrenergic agonists severely suppress the IR2G and cannot be used for sedation for this test. The use of DET did not alter the GR2I, and therefore may be useful for conducting this test in fractious horses., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Five experiments were performed to test the hypothesis that α-melanocyte-stimulating hormone (MSH) is secreted in response to various stressors in horses similar to prolactin, growth hormone, and adrenocorticotropin (ACTH). There was considerable variation in resting concentrations of MSH and in the degree of stimulation in responders; thus all data sets were tested for heterogeneity of variance and corrected for as needed before analysis. In experiment 1, 12 mares were used in a switchback design to test the effect of a 2-minute exercise bout on MSH secretion. Plasma MSH concentrations were constant when mares were not exercised but increased (P < .05) immediately (2 minutes) after exercise and were still elevated 5 minutes later. In experiment 2, six mares were twitched for 2 minutes and six mares were not twitched. Twitching stimulated (P < .05) both MSH and ACTH relative to controls. Experiments 3, 4, and 5 tested the acute effects of intravenous injection of epinephrine at 5 μg/kg of body weight, intravenous injection of 100 μg substance P, and intramuscular injection of 10 mg prostaglandin-F 2α in mares compared to controls (6, 5, and 6 mares per treatment group, respectively). Concentrations of MSH increased (P ≤ .05) after treatment in all three experiments. Plasma concentrations of ACTH also increased (P < .01) after administration of epinephrine and prostaglandin-F 2α in experiments 3 and 5; plasma ACTH was not measured in experiment 1 or 4 because we have previously reported that exercise and substance P stimulate plasma ACTH concentrations. As hypothesized, MSH is secreted in response to various stimuli similar to that observed previously for prolactin, growth hormone, and ACTH., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The objectives of this study were to determine the concentrations of reproductive and metabolic hormones during the peripubertal period and to assess their relationship with testicular development and body fat deposition. Blood samples were collected from 23 healthy Standardbred colts every four weeks for twelve months. Colts were weighed monthly, and percent of body fat and testicular volume estimated by ultrasound. Onset of puberty was determined as the month when testosterone was two standard deviations above the previous mean. Plasma FSH, LH, leptin, estradiol-17β, androstenedione, IGF-1, insulin, inhibin-A, and inhibin-B were analyzed for a seven month peripubertal period. Spring born Standardbred colts underwent puberty at 13 months of age; onset of puberty coincided with exponential testicular growth but did not coincide with an increase in cutaneous body fat deposition or leptin (p > 0.05). Plasma inhibin-B concentrations were significantly increased in the postpubertal period (p < 0.05), but no increase was seen in inhibin-A, androstenedione, FSH, LH, or estradiol-17β. In conclusion, the rise in testosterone and subsequent onset of puberty coincides with rapid testicular growth but is not correlated with an increase in gonadotropins, IGF-1, cutaneous body fat or leptin in the horse., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Four experiments assessed factors affecting prolactin responses to sulpiride administration in horses. Experiment 1 compared the efficacy of the (-) enantiomer of sulpiride to that of the commonly used (+/-) racemic mixture. Mares were used in an 8 × 8 Latin square to compare the prolactin responses to four doses of levosulpiride to four corresponding doses of the racemic mixture at twice the dose. Responses at each dose indicated equal and similar (P > .1) responses. Experiment 2 compared the efficacy of 1 gram of orally administered racemic sulpiride to 100 mg of intramuscularly injected sulpiride in oil in mares primed with 50 mg of estradiol cypionate (ECP). Prolactin responses in groups receiving sulpiride were robust but similar in magnitude with minor differences in timing. In experiment 3, ECP-primed geldings received subcutaneous injections of 1.8 grams racemic sulpiride in vegetable shortening in one of three sites: the neck, the back below the withers, or the lower girth region; control geldings received no sulpiride. Prolactin responses to sulpiride lasted a minimum of 96 hours. In experiment 4, prolactin responses to 3 g of racemic sulpiride in vegetable shortening were compared to similar injections (3 g) in 5 mL of sucrose acetate isobutyrate (SAIB; SucroMate) or just SAIB (control) in ECP-primed geldings. Controls had no prolactin response to SucroMate, whereas both treatment groups had extended prolactin responses lasting at least 10 days. It is concluded that prolactin responses to sulpiride in horses can be greatly extended by using hydrophobic vehicles like vegetable shortening or SAIB., (Copyright © 2018 Elsevier Inc. All rights reserved.)

This experiment assessed the hormonal production, secretory aspects, and changes in luteinizing hormone (LH) receptor gene expression of early induced ovulatory-sized follicles relative to the first ovulatory-sized follicles occurring naturally in the spring. Anovulatory mares were treated on January 21 with (1) 50 mg of estradiol cypionate (ECP, n = 8) alone or (2) with ECP followed by two 3-g sulpiride injections (n = 8), 5 and 12 days later. Half of each group also received complete follicle ablation via transvaginal aspiration before ECP treatment. Ovaries were scanned regularly until detection of a 32-35 mm follicle; follicular fluid was recovered via aspiration for analysis of hormonal concentrations. Blood was collected regularly to characterize plasma prolactin, LH, follicle stimulating hormone, progesterone, and estradiol concentrations. Mean date to first 35-mm follicle was earlier (P < .05) in sulpiride-treated mares: five of eight (63%) responded within 28 days of the first sulpiride injection. Ablation did not affect ovarian response. Plasma prolactin was stimulated (P < .0001) in ECP-sulpiride-treated mares for 16 days but did not dictate ovarian response. Estradiol stimulated plasma LH (P < .05), which was higher (P < .05) in treated mares that responded. There was no effect of treatment or ablation on follicular fluid concentrations of estradiol, progesterone, leptin, or insulin-like growth factor 1 or on LH receptor gene expression. These latter similarities indicate that ECP-sulpiride early induced follicles have apparently reached a degree of maturity equivalent to naturally occurring ovulatory-sized follicles later in the spring., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The aim of the current project was to characterize the luteal vascularity and the plasma concentrations of progesterone (P4), prolactin (PRL) and 13,14-dihydro-15-keto-PGF 2α (PGFM) in mares with luteal disturbances during early and mid-diestrus. In Experiment 1, twenty-one mares were treated with 2 mL of 0.9% NaCl, or 1 mg Dinoprost, or 10 mg Dinoprost on day two after ovulation (Control-D2, 1/10PGF-D2 and PGF-D2 groups, respectively; n = 7 mares/group). In Experiment 2, similar treatments were performed eight days post-ovulation using a different cohort of 21 mares (Control-D8, 1/10PGF-D8 and PGF-D8 groups, respectively; n = 7 mares/group). Blood samples were collected hourly and power-Doppler examinations of the corpus luteum (CL) were performed every 6 h from H0 (moment immediately before treatment) to H48. Data collection was also done once a day from D0 (day of ovulation) to D20. In Experiment 1, the PGF-D2 and 1/10PGF-D2 groups had lower increase of plasma concentration of P4 until H48 and reduced maximum P4 concentrations on D8-D11 than mares from the Control-D2 group. However, no differences among groups were detected for luteal vascularity during early and mid-diestrus. In Experiment 2, complete and partial luteolysis were detected in mares from the PGF-D8 and 1/10PGF-D8 groups, respectively. Luteal vascularity and plasma P4 concentrations differed among Control-D8, PGF-D8 and 1/10PGF-D8 groups on H48. Partially regressed CLs (1/10PGF-D8 group) generated more Doppler signals than completed regressed CLs (PGF-D8 group) between D10 and D13. In both experiments, a transient increase in PRL activity was observed in parallel to the PGFM pulse in mares receiving 1 or 10 mg Dinoprost. The use of prostaglandin on D2 at conventional or 1/10 of the dose impaired the luteal development in mares. Moreover, the low dose of prostaglandin lead to partial regression of mature CLs. The blood supply was reduced in partially regressed CLs, but not in CLs undergoing impaired luteogenesis., (Published by Elsevier Inc.)

The objective of the current study was to evaluate the effects of deslorelin and hCG, two ovulation-inducing therapies, on LH surge and follicle vascularity in mares. Thirty mares were either treated with 1.5 mg IM of deslorelin, 2,500 IU IV of hCG or 2 mL IM of NaCl 0.9% (GnRH, hCG and Saline groups, respectively). Power-flow Doppler examination and blood collection were performed every hour during the first 12 hours after treatment (H0) and every six hours between hours 12 (H12) and 30 (H30) after treatment. Moreover, endpoints were evaluated every hour through the last six hours before ovulation (OV-6 to OV-1). In GnRH group, plasma LH concentration progressively increased (P < 0.001) during the first 6 hours after treatment and remained high (P > 0.1) until OV-1. A significant increase in LH concentrations was first detected (P < 0.05) at 24 hours after treatment in hCG group, while no changes (P > 0.1) on LH levels were found during H0-H30 and between OV-6 and OV-1 in the Saline group. Independent of the treatment, significant variations on the percentage of the follicle wall with Doppler signals were not observed (P > 0.1) throughout the entire experiment. A weak correlation between the preovulatory follicle vascularity and the plasma LH concentration was found in GnRH, hCG and Saline groups (r = +0.29, +0.29 and -0.23, respectively; P ˂ 0.0001). These results described for the first time the immediate and continuous pituitary response to ovulation-inducing therapy with injectable deslorelin. Moreover, spontaneous and induced ovulations were not preceded by an increased follicle vascularity, which differs from previous reports in large animals., (Published by Elsevier Inc.)

The present experiment was divided into two studies to investigate the effect of age and endometrial degeneration on uterine blood flow of mares throughout the immediate post-breeding period. In study 1, uterine blood flow was characterized in mares (n = 7 mares/group) with minimal, moderate or severe endometrial degenerative changes (GI, GII and GIII, respectively). In study 2, the effect of age was investigated using young (≤ 6 years) and old (≥ 15 years) mares (n = 7 mares/group). Uterine vascular perfusion and mesometrial pulsatility index (PI) were evaluated every hour from H0 (moment immediately before AI) to H12. In study 1, a pronounced and transitory increase on uterine vascular perfusion was detected (P < 0.001) between H1 and H3 in the three endometrial groups. In addition, GIII mares had greater (P < 0.05) mesometrial PI than GI mares during the post-breeding period, denoting reduced uterine blood flow. In study 2, a transitory increase on uterine vascular perfusion was also observed in both age groups during the first hour after mating. However, mesometrial PI of young and old mares was similar (P > 0.05) and constant (P > 0.7) through the first 12h after AI. Results demonstrated, for the first time, the immediate changes on uterine vascular perfusion and mesometrial PI in response to semen infusion. Moreover, reduced blood flow of the uterus during the post-breeding period was strongly associated with endometrial degenerative changes in mares, regardless of age., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Six insulin-sensitive and 6 insulin-insensitive mares were used in a replicated 3 by 3 Latin square design to determine the pituitary hormonal responses (compared with vehicle) to sulpiride and thyrotropin-releasing hormone (TRH), 2 compounds commonly used to diagnose pituitary pars intermedia dysfunction (PPID) in horses. Mares were classified as insulin sensitive or insensitive by their previous glucose responses to direct injection of human recombinant insulin. Treatment days were February 25, 2012, and March 10 and 24, 2012. Treatments were sulpiride (racemic mixture, 0.01 mg/kg BW), TRH (0.002 mg/kg BW), and vehicle (saline, 0.01 mL/kg BW) administered intravenously. Blood samples were collected via jugular catheters at -10, 0, 5, 10, 20, 30, 45, 60, 90, and 120 min relative to treatment injection. Plasma ACTH concentrations were variable and were not affected by treatment or insulin sensitivity category. Plasma melanocyte-stimulating hormone (MSH) concentrations responded (P < 0.01) to both sulpiride and TRH injection and were greater (P < 0.05) in insulin-insensitive mares than in sensitive mares. Plasma prolactin concentrations responded (P < 0.01) to both sulpiride and TRH injection, and the response was greater (P < 0.05) for sulpiride; no effect of insulin sensitivity was observed. Plasma thyroid-stimulating hormone (TSH) concentrations responded (P < 0.01) to TRH injection only and were higher (P < 0.05) in insulin-sensitive mares in almost all time periods. Plasma LH and FSH concentrations varied with time (P < 0.05), particularly in the first week of the experiment, but were not affected by treatment or insulin sensitivity category. Plasma GH concentrations were affected (P < 0.05) only by day of treatment. The greater MSH responses to sulpiride and TRH in insulin-insensitive mares were similar to, but not as exaggerated as, those observed by others for PPID horses. In addition, the reduced TSH concentrations in insulin-insensitive mares are consistent with our previous observation of elevated plasma triiodothyronine concentrations in hyperleptinemic horses (later shown to be insulin insensitive as well)., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Four experiments were conducted 1) to assess the use of glucose responses to insulin injections as a means of estimating insulin sensitivity in horses and 2) to compare the insulin sensitivities of normal horses vs. those displaying hyperleptinemia (HL). In Exp. 1, HL mares and geldings (n = 4 each) and 4 mares and geldings with normal leptin concentrations (NL) were injected intravenously with 20 and 100 mU/kg of BW of bovine insulin on 2 separate occasions in December 2008. In Exp. 2, the experimental protocol was repeated in late April 2009. In Exp. 1, the 20 mU/kg of BW dose of insulin caused a greater (P < 0.05) decline in glucose concentrations in NL mares and geldings compared with HL horses. The response of HL mares to the 100 mU/kg of BW dose was less (P < 0.05) than for the other groups. In Exp. 2, responses of all groups to the 20 mU/kg of BW dose were small and similar among groups (P > 0.1), whereas the greater dose revealed differences (P < 0.05) in sensitivity among groups consistent with those observed with the smaller dose in Exp. 1. Experiment 3 was conducted in June and July of 2009 to further examine the dose-response relationship in mares of potentially different insulin sensitivities in an attempt to standardize the approach for studying a wide range of sensitivities. Recombinant human insulin was used at doses of 8, 20, 50, and 125 mU/kg of BW, as needed, to estimate (by linear regression) the dose of insulin causing a 50% decline in glucose concentrations (ED50). Five mares each of reduced leptin concentrations (LL) and small BCS (3 to 5), LL and larger BCS (6 to 7.5), and increased leptin concentrations and increased BCS were studied. The ED50 was similar (P > 0.1) for LL mares, regardless of BCS, and was less (P < 0.01) than for mares with increased leptin concentrations. It was concluded that a dose of 50 mU/kg of BW of recombinant human insulin could be used safely to start the dose-response curve; smaller or larger doses could then be applied as appropriate to get sufficient data for estimation of ED50. Experiment 4, conducted in October of 2009, assessed the repeatability of the estimates for ED50 obtained in Exp. 3. Six mares with LL vs. increased leptin concentrations received the 50 mU/kg dose of insulin; appropriate larger or smaller doses were used to obtain estimates of ED50. Estimates obtained were highly correlated (r = 0.91) with those obtained in Exp. 3, with an average within-mare CV of 8.9%; this is equal to or better than the repetabilities of the currently used methods of assessing insulin sensitivity in horses. It was concluded that hyperleptinemic horses, which are also hyperinsulinemic and have exaggerated insulin responses to glucose injection, are indeed less sensitive to insulin than normal horses with reduced leptin concentrations of the same body condition.

Mares and geldings in good body condition selected for hyperleptinemia vs. normal leptin concentrations were studied to determine whether the hyperleptinemic condition affected various characteristics of the hematologic and hormonal systems after a challenge with lipopolysaccharide endotoxin. Four mares and 4 geldings that were determined to be hyperleptinemic (mean plasma leptin concentrations of 10.0 to 15.5 ng/mL) and 4 mares and 4 geldings with mean plasma leptin concentrations between 2.4 and 5.5 ng/mL were administered Escherichia coli O55:B5 endotoxin (35 ng/kg of BW in 500 mL of saline over a 30-min infusion), or saline only, in pairs in a single-switchback design, with horses and treatments randomly assigned for the first infusion. Physiological variables and blood components were monitored for 24 h after the onset of infusions. Treatments were switched and the second infusions were administered 8 d later. Relative to vehicle infusion, endotoxin infusion increased (P < 0.01) the rectal temperature, heart rate, respiration rate, plasma total protein concentration, and blood packed cell volume; there was an interaction of leptin status, endotoxin treatment, and time for heart rate (P = 0.039), respiration rate (P = 0.018), and plasma total protein concentration (P = 0.054). Blood concentrations of leukocytes, lymphocytes, and neutrophils all decreased (P < 0.001) after endotoxin infusion; there was an interaction (P = 0.0057) between sex and leptin status for blood platelet concentration. Plasma leptin concentrations increased (P = 0.013) after endotoxin infusion in both hyperleptinemic horses and those with reduced leptin concentrations. There were interactions (P < 0.037) of sex with endotoxin treatment and time for plasma concentrations of cortisol and prolactin, whereas plasma GH concentrations were affected (increased; P < 0.001) only by time after infusion. Given that the effects of hyperleptinemia were generally minor, it was concluded that the hyperleptinemic condition, and its associated type-2 diabetic symptoms, has a minimal impact on the components of the hematologic and hormonal systems studied.

Testicular function is under the control of expression and repression of several genes and gene products, and many of these works through Sertoli cells. The capability of Sertoli cells to regulate spermatogenesis is dependent on Sertoli cell functions and Sertoli cell number. Sertoli cell number has long been thought to be stable in adults with no proliferation of Sertoli cells once adult numbers have been reached. However, adult horses do not have stable Sertoli cell numbers, and new studies indicate that adult Sertoli cells can be made to re-enter mitotic phase under certain experimental conditions. This review discusses roles of Sertoli cells in regulation of spermatogenesis and methods for estimating the number of Sertoli cells, in a testis, that overcome the problems (assumptions) associated with the indented, pear-shaped of Sertoli cell nuclei which make it difficult to estimate the volume of individual nuclei. Using several approaches to overcome the problems associated with any one method, the horse is identified as a species in which Sertoli cell number is not fixed, but it fluctuates with season. In addition to Sertoli cell numbers, the functions of Sertoli cells that are very important in signaling and controlling spermatogenesis are discussed. Recent studies have shown that "post-mitotic terminally differentiated Sertoli cells" from adult animals could, under certain conditions, re-enter the cell division cycle. Can seasonal influences be a natural set of conditions to induce the Sertoli cells of the horse testis to seasonally re-enter the cell division cycle and explain the seasonal differences in Sertoli cell number as summarized in this review? Alternatively, can seasonal differences in Sertoli cell number reflect, in the horse to a greater extent, but in adults of most species, the presence of some mitotic-capable Sertoli cells in adults? In any case, both Sertoli cell number and function are important in regulation of spermatogenesis.

We previously reported that a rise in plasma leptin concentrations followed the rise in insulin and glucose in meal-fed horses, whereas horses maintained on pasture had little fluctuations in hormonal patterns. We have also described a hyperleptinemic-hyperinsulinemic condition that occurs in about 30% of our light horse mares of high body condition maintained on pasture. The present experiment was designed to 1) study the effect of 3 common feeding-housing regimens on leptin and other metabolic hormones in mares and 2) determine whether the hyperleptinemic condition interacted with these regimens. Six light horse mares with high body condition (average score = 7) were assigned to 2 simultaneous 3 x 3 Latin squares, 1 with normal mares (leptin = 0.1 to 6 ng/mL) and 1 with mares displaying hyperleptinemia (>10 ng/mL). Three feeding-housing regimens were compared: ad libitum pasture, ad libitum native grass hay in an outdoor paddock, and single morning feedings of a pelleted concentrate and hay at 0700 in a barn. Five days of acclimation to the feeding regimens were followed by a 36-h period of hourly blood collection to characterize the hormonal characteristics. Leptin concentrations were elevated (P < 0.001) in mares predetermined to be hyperleptinemic compared with normal mares, regardless of the feeding regimen. Leptin was greatest (P < 0.01) in mares on pasture and least in mares fed hay. Variations over time (P < 0.01) were present for all hormones and metabolites studied. Glucose and insulin concentrations were greatest (P < 0.01) in mares on pasture, with meal-fed mares exhibiting an immediate rise in plasma concentrations of both after feeding. Mares on hay had low and constant concentrations of glucose, insulin, and leptin, with no apparent fluctuations. Cortisol, prolactin, and IGF-I did not differ with leptin status, whereas GH differed due to feeding-housing regimen (P < 0.02); there was also an interaction of leptin status and feeding-housing regimen for GH concentrations (P = 0.094). It was concluded that 1) estimates of hormonal secretion in horses based on frequent sampling, depending upon the hormone in question, can be profoundly affected by the feeding-housing regimens, and 2) the hyperleptinemic condition persists under differing conditions of feeding-housing.

Previous results from our laboratory indicated that a majority of mares with high body condition scores (BCS) displayed estrous cycles or had considerable follicular activity during the winter. Among these high BCS mares, about 35% of them exhibited a persistent hyperleptinemia and hyperinsulinemia. The current experiment was designed to compare the reproductive characteristics of high BCS mares with hyperleptinemia to those with normal (low) plasma concentrations of leptin during the winter and the first estrous cycle (or the first full cycle encountered for those already cycling). Light horse mares with high BCS (6-8.5) were assigned to groups based on leptin concentrations (8/group): low (<5 ng/mL) and high (>10 ng/mL). Beginning 7 January, mares were assessed every 3d for follicular activity and then daily once a follicle >25 mm was detected. Mares were subsequently monitored through their first and second ovulations. Leptin concentrations remained higher (P<0.001) in mares in the high leptin group over the duration of the experiment. Also, high leptin mares had greater (P<0.0001) insulin response to glucose infusion and a faster (P<0.05) rate of glucose clearance. One mare with high leptin and three mares with low leptin had progesterone concentrations indicative of the presence of a corpus luteum at the onset of the experiment. Plasma concentrations of LH, FSH, and progesterone did not differ between groups (P>0.1) during the first estrous cycle occurring after 7 January. Date of first ovulation after 7 January and interovulatory interval were similar (P>0.1) for the two groups, as were estimates of follicular numbers on the ovaries (small, medium, and large; P>0.1). It is concluded that the perturbations in leptin and insulin secretion observed in some high BCS mares are not associated with alterations in ovarian activity or the estrous cycle during winter and into the period of vernal transition.

Five experiments were performed to evaluate the effects of dexamethasone (DEX), gender, and testosterone on plasma leptin concentrations in horses. In experiment 1, plasma leptin, insulin, glucose, and IGF-1 concentrations were increased (P < 0.01) in stallions following five daily injections of DEX (125 microg/kg BW). In experiment 2, leptin concentrations increased (P < 0.01) in mares, geldings, and stallions following a single injection of DEX, and the response was greater (P < 0.01) in mares and geldings than in stallions. The gender effect was confounded by differences in body condition scores and diet; however, based on stepwise regression analysis, both BCS and gender were significant sources of variation in the best fit model for pre-DEX leptin concentrations (R(2) = 0.65) and for maximum leptin response to DEX (R(2) = 0.75). In experiment 3, in which mares and stallions were pair-matched based on age and body condition and fed similar diets, mares again had higher (P < 0.01) leptin concentrations than stallions after DEX treatment as used in experiment 2. In experiment 4, there was no difference (P > 0.1) in plasma leptin response in mares following four single-injection doses of DEX from 15.6 to 125 microg/kg BW. In experiment 5, treatment of mares with testosterone propionate every other day for 5 days did not alter (P > 0.1) plasma leptin concentrations or the leptin response to DEX. In conclusion, multiple injections of DEX increase leptin concentrations in stallions, as does a single injection in mares (as low as 15.6 microg/kg BW), geldings and stallions. The greater leptin levels observed in mares and geldings relative to stallions were due partially to their greater body condition and partially to the presence of hyperleptinemic individuals; however, even after accounting for body condition and diet, mares still had greater leptin concentrations than stallions after DEX administration. Elevation of testosterone levels in mares for approximately 10 days did not alter leptin concentrations in mares.

Three experiments tested the hypotheses that daily cortisol rhythm, feeding time, and/or insulin infusion affect(s) leptin secretion in stallions. Ten mature stallions received ad libitum hay and water and were fed a grain concentrate once daily at 0700. In Exp. 1, stallions received either a single injection of dexamethasone (125 microg/kg BW i.m.; n = 5) or vehicle (controls; n = 5) at 0700 on d -1. Starting 24 h later, blood samples were collected every 2 h for 36 h via jugular venipuncture. Cortisol in control stallions varied (P < 0.01) with time, with a morning peak and evening nadir; dexamethasone suppressed (P < 0.01) cortisol concentrations. Leptin and insulin were greater (P < 0.01) in the treated stallions, as was the insulin response to feeding (P < 0.01). Leptin in control stallions varied (P < 0.01) in a diurnal pattern, peaking approximately 10 h after onset of eating. This pattern of leptin secretion was similar, although of greater magnitude (P < 0.01), in treated stallions. In Exp. 2, five stallions were fed the concentrate portion of their diet daily at 0700 and five were switched to feeding at 1900. After 14 d on these regimens, blood samples were collected every 4 h for 48 h and then twice daily for 5 d. Cortisol varied diurnally (P = 0.02) and was not altered (P = 0.21) by feeding time. Insulin and leptin increased (P < 0.01) after feeding, and the peaks in insulin and leptin were shifted 12 h by feeding at 1900. In Exp. 3, six stallions were used in two 3 x 3 Latin square experiments. Treatments were 1) normal daily meal at 0700; 2) no feed for 24 h; and 3) no feed and a bolus injection of insulin (0.4 mIU/kg BW i.v.) followed by infusion of insulin (1.2 mIU.kg BW(-1).min(-1)) for 180 min, which was gradually decreased to 0 by 240 min; sufficient glucose was infused to maintain euglycemia. Plasma insulin increased (P < 0.01) in stallions when they were meal-fed (to approximately 150 microIU/mL) or infused with insulin and glucose (to approximately 75 microIU/mL), but insulin remained low (10 microIU/mL or less) when they were not fed. The increases in insulin were paralleled by gradual increases (P < 0.01) in leptin concentrations 3 to 4 h later in stallions fed or infused with insulin and glucose. When stallions were not fed, leptin concentrations remained low. These results demonstrate that feeding time, and more specifically the insulin increase associated with a meal, not cortisol rhythm, drives the postprandial increase in plasma leptin concentrations in horses.

Objective: To determine the effects of levothyroxine sodium (L-T4) on serum concentrations of thyroid gland hormones and responses to injections of thyrotropin-releasing hormone (TRH) in euthyroid horses., Animals: 12 healthy adult mares., Procedure: 8 horses received an incrementally increasing dosage of L-T4 (24, 48, 72, or 96 mg of L-T4/d) for weeks 1 to 8. Each dose was provided for 2 weeks. Four additional horses remained untreated. Serum concentrations of total triiodothyronine (tT3), total thyroxine (tT4), free T3 (fT3), free T4 (fT4), and thyroid-stimulating hormone (TSH) were measured in samples obtained at weeks 0, 2, 4, 6, and 8; 1.2 mg of TRH was then administered i.v., and serum concentrations of thyroid gland hormones were measured 2 and 4 hours after injection. Serum reverseT3 (rT3) concentration was also measured in the samples collected at weeks 0 and 8., Results: Treated horses lost a significant amount of weight (median, 19 kg). Significant treatment-by-time effects were detected for serum tT3, tT4, fT3, fT4, and TSH concentrations, and serum tT4 concentrations were positively correlated (r, 0.95) with time (and therefore dosage) in treated horses. Mean +/- SD serum rT3 concentration significantly increased in treated horses (3.06 +/- 0.51 nmol/L for week 8 vs 0.74 +/- 0.22 nmol/L for week 0). Serum tT3, tT4, fT3, and TSH concentrations in response to TRH injections differed significantly between treated and untreated horses., Conclusions and Clinical Relevance: Administration of levothyroxine sodium increased serum tT4 concentrations and blunted responses toTRH injection in healthy euthyroid horses.

In the present study, follicular fluids of estrous mares treated with saline solution (Control) or nitric oxide synthase (NOS) inhibitors were analyzed for nitric oxide (NO), estradiol-17beta (E2) and progesterone (P4) concentrations before and 36h after administration of human chorionic gonadotropin (hCG). Follicular fluids obtained before (0h) hCG administration from control mares had lower concentrations of NO than those obtained 36h after administration of hCG (58.3+/-17.8 micromol versus 340.4+/-57.7 micromol; P<0.05). A similar pattern was also noted for intrafollicular P4 in control mares, which had lower concentrations of intrafollicular P4 before hCG than 36h post-hCG administration (P<0.05). As expected, E2 concentrations of control follicles sampled before hCG administration were higher than those sampled 36h post-hCG administration (P<0.05). However, the E2 concentrations in follicles of mares treated with the NOS inhibitors N(omega)-nitro-L-arginine methyl ester (L-NAME) or aminoguanidine (AG) did not decrease after hCG administration, unlike those in control mares (P>0.10). In addition, mares treated with NOS inhibitors had lower intrafollicular concentrations of NO and P4 than control mares, both before and after hCG administration (P<0.05). Increased intrafollicular concentrations of NO in control, hCG-stimulated mares provide evidence for the presence of an NO-generating system in the equine preovulatory follicle that is likely upregulated following administration of hCG.

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.

Previous observations from this laboratory indicated that horses with high BCS could have resting plasma leptin concentrations ranging from low (1 to 5 ng/mL) to very high (10 to 50 ng/mL). To study the possible interactions of leptin secretion with other endocrine systems, BCS and plasma leptin concentrations were measured on 36 mares and 18 geldings. From mares and geldings that had a mean BCS of at least 7.5, five with the lowest (low leptin) and five with the highest (high leptin) leptin concentrations were selected. Jugular blood samples were collected twice daily for 3 d from the 20 selected horses to determine average resting hormone concentrations. Over the next 12 d, glucose infusion, injection of thyrotropin-releasing hormone (TRH), exercise, and dexamethasone treatment were used to perturb various hormonal systems. By design, horses selected for high leptin had greater (P < 0.0001) leptin concentrations than horses selected for low leptin (14.1 vs. 2.8 +/- 0.92 ng/mL, respectively). In addition, mares had greater (P = 0.008) leptin concentrations than geldings. Horses selected for high leptin had lower (P = 0.027) concentrations of GH but higher (P = 0.0005) concentrations of insulin and thriiodothyronine (T3) than those selected for low leptin. Mares had greater (P = 0.0006) concentrations of cortisol than geldings. There was no difference (P > 0.10) in concentrations of IGF-1, prolactin, or thyroid-stimulating hormone (TSH). Horses selected for high leptin had a greater (P = 0.0365) insulin response to i.v. glucose infusion than horses selected for low leptin. Mares had a greater (P = 0.0006) TSH response and tended (P = 0.088) to have a greater prolactin response to TRH than geldings; the T3 response was greater (P = 0.047) in horses selected for high leptin. The leptin (P = 0.0057), insulin (P < 0.0001), and glucose (P = 0.0063) responses to dexamethasone were greater in horses selected for high leptin than in those selected for low leptin. In addition, mares had a greater (P < 0.0001) glucose response to dexamethasone than geldings. Cortisol concentrations were decreased (P = 0.029) by dexamethasone equally in all groups. In conclusion, differences in insulin, T3, and GH associated with high vs. low leptin concentrations indicate a likely interaction of these systems with leptin secretion in horses and serve as a starting point for future study of the cause-and-effect nature of the interactions.

Three experiments were performed to test the following hypotheses: 1) stallions and/or progesterone-estradiol-treated geldings could serve as models for the effects of a single implant of the GnRH analog, deslorelin acetate, on LH and FSH secretion by mares; and 2) multiple implants of deslorelin acetate could be used as a means of inducing ovarian atrophy in mares for future study of the mechanisms involved in the atrophy observed in some mares after a single implant. In Exp. 1, nine light horse stallions received either a single deslorelin implant (n = 5) or a sham injection (n = 4) on d 0. In Exp. 2, 12 geldings received daily injections of progesterone on d -20 through -4, followed by twice-daily injections of estradiol on d -2 to 0. On the morning of d 0, geldings received either a single deslorelin implant (n = 6) or a sham injection (n = 6). Daily injections of progesterone were resumed on d 2 through 15. In Exp. 1, plasma LH and FSH were elevated (P < 0.05) in the treatment group relative to controls at 4, 8, and 12 h after implant insertion. In the treated stallions, FSH was decreased (P < 0.05) on d 3 to 13, and LH was decreased on d 6 to 13. In Exp. 2, plasma LH and FSH were elevated (P < 0.05) at 4,8, and 12 h after deslorelin implant insertion. Plasma LH was suppressed (P < 0.05) below controls on d 2 to 7, 9, and 11 to 15; plasma FSH was suppressed (P < 0.05) on d 4 to 15. In Exp. 3, 21 mares were used to determine whether multiple doses of deslorelin would cause ovarian atrophy. Mares received one of three treatments: 1) sham injections; 2) three implants on the first day; or 3) one implant per day for 3 d (n = 7 per group). Treatment with multiple implants increased (P < 0.05) the interovulatory interval by 14.8 d and suppressed (P < 0.01) LH and FSH concentrations for approximately 25 d; no mare exhibited ovarian atrophy. In conclusion, after an initial short-term increase in LH and FSH secretion, deslorelin implants caused long-term suppression of both gonadotropins in stallions as well as in geldings treated with progesterone and estradiol to mimic the estrous cycle. It is likely that either of these models may be useful for further study of this suppression in horses. Although multiple implants in mares suppressed gonadotropin secretion longer than a single implant, the lack of ovarian atrophy indicates that the atrophy observed after a single implant in previous experiments was likely due to the susceptibility of individual mares.

Reasons for Performing Study: There exists a need for better diagnostic tests to characterise thyroid disease in horses. Currently available diagnostic tests fail to differentiate between thyroid gland disorders and thyroid abnormalities resulting from pituitary or hypothalamic problems., Objectives: To evaluate the effects of treatment with propylthiouracil (PTU) and bromocryptine (BROM) on serum concentrations of triiodothyronine (T3), thyroxine (T4), reverse T3 (rT3) and equine thyroid-stimulating hormone (e-TSH, thyrotrophin) in mature horses., Methods: Healthy mature horses were treated using either PTU or BROM for 28 days. The effect of treatment on the thyroid axis was assessed by measuring T3, T4, rT3 and e-TSH before and at +14 and +28 days. The effect of PTU and BROM on the response of T3, T4, rT3 and e-TSH to thyrotrophin-release hormone (TRH) administration was also assessed before and at +14 and +28 days of treatment., Results: Treatment with PTU led to a significant reduction in serum concentrations of T3, T4 and rT3 on Day 28 and increase of e-TSH on Day 28 (P < 0.05). Treatment with BROM did not cause any measurable effect on serum concentrations of T3, T4, rT3 or e-TSH. The percentage increment by which serum concentration of T4, T3 and e-TSH increased following stimulation with TRH was decreased by treatment with PTU for 28 days (P < 0.05) but were not affected by treatment with BROM for 28 days., Conclusions: These results suggest that 1) treatment with PTU may be used in horses as a model of primary hypothyroidism; 2) the use of BROM as a model of secondary hypothyroidism in horses is not supported; and 3) e-TSH assay deserves further investigation for the clinical diagnosis of thyroid axis dysfunction in horses., Potential Relevance: Propylthiouracil effectively causes primary hypothyroidism. There is substantial variability between horses with respect to their sensitivity to this substance when administered orally. Further studies pertaining to the characterisation of equine thyroid disorders are warranted and the use of both PTU for the experimental induction of primary hypothyroidism and e-TSH for the diagnostic characterisation of thyroid disorders in horses should be considered.

In experiment 1, nine light horse geldings (three 3 x 3 Latin squares) received dexamethasone (DEX; 125 microg/kg BW, i.m.), glucose (0.2 g/kg BW, i.v.), or nothing (control) once per day for 4 days. DEX increased (P < 0.001) glucose, insulin, and leptin concentrations and resulted in a delayed increase (P < 0.001) in IGF-I concentrations. In experiment 2, mares were similarly treated with DEX (n = 6) or vehicle (n = 6). DEX again increased (P < 0.01) glucose, insulin, and leptin concentrations; the delayed elevation in IGF-I concentrations occurred on day 10, 12, and 19, relative to the first day of treatment. In experiment 3, six light horse geldings received either 200 IU of adrenocorticotropin (ACTH) i.m. or vehicle twice daily for 4 days. ACTH increased (P < 0.001) cortisol concentrations. Further, ACTH resulted in increases (P < 0.01) glucose, insulin, and leptin concentrations. In experiment 4, plasma samples from four light horse stallions that were fed 6-n-propyl-2-thiouracil (PTU) at 6 mg/kg BW for 60 days to induce hypothyroidism were compared to samples from control stallions. On day 52, stallions receiving PTU had lower concentrations of thyroxine (P < 0.05) and triiodothyronine (P < 0.01) and higher (P < 0.01) concentrations of TSH. Leptin concentrations were higher (P < 0.01) in PTU-fed stallions from day 10 through 52. In conclusion, circulating concentrations of leptin in horses was increased by administering DEX. Treatment with ACTH increased cortisol and resulted in lesser increases in leptin, glucose, and insulin. In addition, PTU feeding results in lesser increases in leptin concentrations., (Copyright 2002 Elsevier Science Inc.)

Mares that had previously been fed to attain body condition scores (BCS) of 7.5 to 8.5 (high) or 3.0 to 3.5 (low) were used to determine the interaction of BCS with the responses to 1) administration of equine somatotropin (eST) daily for 14 d beginning January 20 followed by administration of GnRH analog (GnRHa) daily for 21 d and 2) 4-d treatment with dexamethasone later in the spring when mares in low BCS had begun to ovulate. The majority of mares with high BCS continued to cycle throughout the winter, as evidenced by larger ovaries (P < 0.002), more corpora lutea (P < 0.05), greater progesterone concentrations during eST treatment (P < 0.04), and more (P < 0.05) large- and medium-sized follicles. Treatment with eST alone or in combination with GnRHa had no effect (P > 0.05) on ovarian activity or ovulation. Plasma leptin concentrations were greater (P < 0.002) in mares with high BCS; however, there was no effect (P > 0.10) of eST treatment. Plasma IGF-I concentrations were greater (P < 0.0001) in mares treated with eST compared with mares given vehicle, and mares with high BCS had greater IGF-I (P < 0.02) and LH concentrations (P < 0.02) than mares with low BCS. Plasma leptin concentrations in mares with high BCS were increased (P < 0.001) within 12 h of dexamethasone treatment; the leptin response (P < 0.001) in mares with low BCS was greatly reduced (P < 0.001) and transient. Glucose and insulin concentrations also increased (P < 0.0001) after dexamethasone treatment in both groups, and the magnitude of the response was greater (P < 0.0001) in mares with high BCS than in mares with low BCS. In summary,low BCS in mares was associated with a consistent seasonal anovulatory state that was affected little by eST and GnRHa administration. In contrast, all but one mare with high BCS continued to experience estrous cycles and(or) have abundant follicular activity on their ovaries. The IGF-I response to eST treatment was also reduced in mares with low BCS, as was the basal leptin concentration and leptin response to dexamethasone. Although low BCS and leptin concentrations were associated with inactive ovaries during winter and early spring, mares with low BCS eventually ovulated in April and May while leptin concentrations remained low.

The present experiment characterized the pituitary responsiveness to exogenous GnRH in the first 10 d after ovulation following commercially available deslorelin acetate implantation at the normal dosage for hastening ovulation in mares. Twelve mature, cyclic mares were assessed daily for estrus and three times weekly for ovarian activity starting May 1. Mares achieving a follicle at least 25 mm in diameter or showing signs of estrus were checked daily thereafter for ovarian characteristics. When a follicle >30 mm was detected, mares were administered either a single deslorelin acetate implant or a sham injection and then assessed daily for ovulation. On d 1, 4, 7, and 10 following ovulation, each mare was challenged i.v. with 50 microg GnRH, and blood samples were collected to characterize the LH and FSH responses. The size of the largest follicle on the day of treatment did not differ (P = 0.89) between groups. The number of days from treatment to ovulation was shorter (P < 0.001) by 2.0 d for the treated mares indicating a hastening of ovulation. The size of the largest follicle present on the days of GnRH challenge was larger in the treated mares on d 1 (P = 0.007) but smaller on d 10 (P = 0.02). In addition, the interovulatory interval was longer (P = 0.036) in the treated mares relative to controls by 4.4 d. Concentrations of FSH in plasma of the treated mares were lower (P < 0.05) than control concentrations from d 3 to 12; LH concentrations in the treated mares were lower (P < 0.05) relative to controls on d 0 to 5, d 7, and again on d 20 to 23. Progesterone values were the same (P = 0.99) for both groups from 2 d before ovulation though d 23. There was an interaction of treatment, day, and time of sampling (P < 0.001) for LH and FSH concentrations after injection of GnRH. Both the LH and FSH responses were suppressed (P < 0.009) in the treated mares relative to controls on d 1, 4, and 7; by d 10, the responses of the two groups were equivalent. In conclusion, deslorelin administration in this manner increased the interovulatory interval, consistently suppressed plasma LH and FSH concentrations, and resulted in a complete lack of responsiveness of LH and FSH to GnRH stimulation at the dose used during the first 7 d after the induced ovulation. Together, these results are consistent with a temporary down-regulation of the pituitary gland in response to deslorelin administered in this manner.

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.

Two experiments compared the efficacies of different treatment frequencies for recombinant equine somatotropin (eST). In Experiment 1, five geldings received daily injections of eST at 20 microg/kg of body weight, and five received every-other-day injections at 40 microg/kg of body weight, for a total of 30 days. Plasma glucose (P=0.0001), insulin (P=0.0135), and non-esterified fatty acid (NEFA, P=0.0001) concentrations increased, and plasma urea nitrogen (PUN) concentrations decreased (P=0.0001), in both groups, and only minor differences (P<0.05) occurred between the two groups. Insulin-like growth factor-I (IGF-I) concentrations increased (P=0.0001) in both groups over time, and were higher (P<0.05) after day 2 in geldings treated daily. Endogenous somatotropin (ST) response to secretagogue was inhibited (P<0.05) in geldings receiving daily injections relative to those receiving every-other-day injections. In Experiment 2, 16 geldings were allotted to four groups of four. A control group received daily saline injections, and the other three groups received eST at 20 microg/kg of body weight daily as a single injection, two injections (every 12h), or four injections (every 6h), for a total of 14 days. Plasma IGF-I and insulin concentrations increased (P<0.05) in all groups receiving eST, with the responses being proportional to injection frequency. In contrast, PUN concentrations decreased (P<0.05) in all groups equally. In conclusion, the efficacy of daily versus every-other-day injections of eST depends upon the response to be measured, and for IGF-I concentrations, the every-other-day regimen was not acceptable. Injection frequencies greater than once daily were more efficacious for IGF-I and insulin concentrations, but not for PUN concentrations. Thus, the optimum injection regimen for any new application for eST cannot simply be inferred from other biological responses, and will need to be determined empirically.

A series of experiments was performed to determine the factor(s) responsible for an apparent inhibition of GH secretion in mares administered the GH secretagogue EP51389 in combination with GnRH, thyrotropin-releasing hormone (TRH), and sulpiride. Experiment 1 tested the repeatability of the original observation: 10 mares received EP51389 at 10 microg/kg BW; five received TRH (10 microg/kg BW), GnRH (1 microg/kg BW), and sulpiride (100 microg/kg BW) immediately before EP51389, and five received saline. The mixture of TRH, GnRH, and sulpiride reduced (P = 0.0034) the GH response to EP51389, confirming the inhibitory effects. Experiment 2 tested the hypothesis that sulpiride, a dopamine antagonist, was the inhibitory agent. Twelve mares received EP51389 as in Exp. 1; six received sulpiride before EP51389 and six received saline. The GH responses in the two groups were similar (P > 0.1), indicating that sulpiride was not the inhibitory factor. Experiment 3 tested the effects of TRH and(or) GnRH in a 2 x 2 factorial arrangement of treatments. Three mares each received saline, TRH, GnRH, or the combination before EP51389 injection. There was a reduction (P < 0.0001) in GH response in mares receiving TRH, whereas GnRH had no effect (P > 0.1). Given those results, Exp. 4 was conducted to confirm that TRH was inhibitory in vivo as opposed to some unknown chemical interaction of the two compounds in the injection solution. Twenty mares received TRH or saline and(or) EP51389 or saline in a 2 x 2 factorial arrangement of treatments. Injections were given separately so that the two secretagogues never came in contact before injection. Again, TRH reduced (P < 0.0001) the GH response to EP51389. In addition, TRH and EP51389 each resulted in a temporary increase in cortisol concentrations. Experiment 5 tested whether TRH would alter the GH response to GHRH itself. Twelve mares received porcine GHRH at 0.4 microg/kg BW; six received TRH prior to GHRH and six received saline. After adjustment for pretreatment differences between groups, the GHRH-induced GH response was completely inhibited (P = 0.068) by TRH. Exp. 6 was a repeat of Exp. 5, except geldings were used (five per group). Again, pretreatment with TRH inhibited (P < 0.0001) the GH response to GHRH. In conclusion, TRH inhibits the GH response not only to EP51389 but also to GHRH in horses, and in addition to its known secretagogue action on prolactin and TSH it may also stimulate ACTH at the dosage used in these experiments.

This experiment assessed the effects of 12 mo of daily treatment of young horses with recombinant equine somatotropin (eST) on 1) carcass and internal organ traits at necropsy and 2) residual effects in live horses for 60 d after cessation of treatment. Seven horses received eST daily at 20 microg/kg BW; seven others received vehicle (controls). Four horses from each group were killed at the end of treatment. There were few effects of eST treatment on hematologic assessments or histopathologic evaluations of internal organs. Treatment with eST increased the weights of the right adrenal gland (P = 0.090), left (P = 0.085) and right (P = 0.013) kidneys, liver (P = 0.012), tended to inrease the weights of pancreas (P = 0.082), spleen (P = 0.008), and heart (P = 0.102), and decreased (P = 0.032) somatotropin (ST) content in the adenohypophysis. Loin-eye area at the 10th rib was also greater (P = 0.01) in eST-treated horses than in controls. There was no difference (P > 0.15) between groups in left adrenal, brain, parathyroid glands, or thyroid gland weights or in 10th-rib fat thickness. In the remaining two control and three eST-treated horses (one control horse died), plasma IGF-I concentrations were higher (P = 0.001) in treated animals through d 6 after cessation of treatment and then dropped precipitously. Insulin concentrations in treated animals tended to be elevated (P = 0.08) only on d 0. There was a treatment x day interaction (P = 0.04) for plasma urea nitrogen levels, which increased in treated horses. A decrease (P < 0.05) in BW in the treated animals was observed by 21 d after treatment. There was no difference (P > 0.15) in insulin or glucose response to glucose tolerance tests given on d 0 through 60 after cessation of treatment. Overall ST response to secretagogue was reduced (P < 0.05) in eST-treated horses compared with controls. In summary, long-term treatment of growing horses with eST decreased endogenous ST response to secretagogue and increased plasma IGF-I concentrations and many internal organ weights but had little effect on hematologic or histopathologic characteristics at necropsy. The effects on IGF-I concentrations were lost within 6 d, and BW in treated horses decreased within 3 wk after cessation of treatment.

Sixteen seasonally anovulatory mares were randomly allotted to two groups and injected daily with either sulpiride (1 mg/kg body weight) or vehicle from 14 January to 14 February. Sulpiride administration increased daily plasma prolactin concentrations (P < 0.05), although the prolactin response during the 6 h following sulpiride injections decreased markedly from the 1st to the 6th day of treatment (treatment by day, P < 0.0001). Plasma concentrations of LH and FSH were not affected by treatment (P > 0.1). Injection of GnRH and TRH on 15 February showed that the response of plasma prolactin to secretagogue was increased in sulpiride-treated mares (P < 0.005), while there was no effect (P > 0.1) of sulpiride treatment on the response of LH or FSH. Both treatment groups had similar changes in numbers of follicles 10-19 and > or = 20 mm during the experiment (P > 0.1). Similarly, the mean change in maximal follicular size was not affected by treatment (P > 0.9). No mare ovulated during the study, and plasma progesterone concentrations were similar in both groups (P > 0.1), always at levels < 1 ng/ml. Hairshedding increased with time in all mares (P < 0.001) and was increased by sulpiride injections (P = 0.09). It was concluded that sulpiride administration to seasonally anovulatory mares under the conditions of our experiment increased daily plasma prolactin levels but did not stimulate gonadotropin secretion or ovarian activity.

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.

Fourteen foals between 4 and 4.5 mo of age were used to determine the effects of 12 mo of daily treatment with equine somatotropin (eST) on growth, metabolic, and hormonal characteristics. The foals were paired by sex, type, and lineage, and one of each pair was administered eST daily at 20 microg/kg of BW. Body weights, body measurements, and assessments of glucose tolerance and feedback effects on endogenous somatotropin (ST) secretion were made routinely. Treatment with eST did not alter (P > 0.10) BW, height at withers, length of body, widths of chest and rump, heart girth, length of head, front or rear cannon lengths, front or rear cannon circumferences, gaskin circumference, or skin thickness, even though plasma IGF-I concentrations were doubled (P = 0.012). Glucose concentrations were higher (P = 0.03) in treated horses before glucose infusion; there was no difference (P > 0.10) in the glucose response to infusion. The insulin response to glucose infusion in the treated horses was generally higher (P = 0.0069) than in controls. Endogenous ST secretion in response to a ST secretagogue was reduced (P = 0.0001) in foals treated with eST in all months. The prolactin and thyroid-stimulating hormone responses to thyrotropin-releasing hormone on June 1 were not affected (P > 0.10) by treatment. In conclusion, daily treatment of growing horses with eST for 12 mo at the recommended dose altered the hormonal and metabolic characteristics known to be affected by ST but did not alter the growth characteristics of the animals.

Thirty-nine adult light horse mares, geldings, and stallions were used in two experiments to assess the pituitary hormone and insulin responses to infusions of arginine, aspartic acid, lysine, glutamic acid, and N-methyl-D,L-aspartate (NMA). In Exp. 1, 27 horses were assigned to one of three infusion treatments: 1) physiological saline (1 L); 2) 2.855 mmol of arginine/kg BW in 1 L of water; or 3) 2.855 mmol of aspartic acid/kg BW in 1 L of water. In Exp. 2, 12 horses were assigned, in a multiple-square 4 x 4 Latin square design, to one of four infusion treatments: 1) 2 mL of saline/kg BW; 2) 2.855 mmol of lysine/kg BW in water; 3) 2.855 mmol of glutamic acid/kg BW in water; or 4) 1 mg of NMA/kg BW in water. In Exp. 1, an acute (within 20 min) release of growth hormone (GH) was induced (P = 0.002) by aspartic acid. In contrast, acute release of prolactin (P = 0.001) and insulin (P = 0.002) was induced only by arginine; moreover, the arginine effect on insulin was present only in mares (P = 0.011). In Exp. 2, an acute release of GH was induced (P = 0.001) by glutamic acid and NMA. In males, the glutamic acid-induced GH release was greater than that of NMA; in mares, the NMA-induced GH release was greater than that of glutamic acid (P = 0.069). Both lysine and glutamic acid induced (P = 0.001) acute release of prolactin, whereas an acute release of insulin was elicited (P = 0.002) only by lysine. The NMA-induced LH response was due almost entirely to the response in mares and stallions (P = 0.016), and the NMA-induced FSH release was due almost entirely to the response in mares (reproductive status effect; P = 0.004). In the horse, aspartic acid, glutamic acid, and NMA seem to stimulate GH release; arginine and lysine seem to stimulate prolactin and insulin release; and NMA seems to stimulate LH and FSH release. It seems that N-methyl-D-aspartate glutamate receptors are involved in controlling GH, LH, and FSH secretion, whereas other mechanisms are involved with prolactin secretion. These results also indicate that gonadal steroids interact with amino acid-induced pituitary hormone release in adult horses.

Development of a controlled release formulation of gonadotropin releasing hormone that would stimulate a LH surge capable of reducing the time span of ovulations would greatly benefit reproductive management because a single timed insemination could be used. A dose-response study was conducted to determine if Deslorelin, a potent gonadotropin releasing hormone analogue, delivered via the SABER system, a biodegradable controlled release system, would stimulate an ovulatory-like LH surge in the pig. Twenty ovariectomized gilts, approximately 200 d old and 100 kg body weight (BW), received estradiol benzoate (15 microg/kg BW im) and 48 h later, the gilts were given deslorelin at 0, 12.5, 25.0, 50.0 or 100.0 microg im (n = 4 each treatment group). Compared to controls, mean blood deslorelin concentrations were still elevated at 30 h after deslorelin. Mean deslorelin magnitude, area under the curve and duration were sequentially greater (P<0.05) in a dose-dependent sequence. Compared to controls, serum LH concentrations were elevated (P<0.05) for 6 to 12 h after deslorelin. A dose-response relationship was absent for all parameters of LH secretion. Magnitude of the serum LH response was greatest (P<0.05) in the 12.5 microg and 50.0 microg groups, whereas area under the curve was lower (P<0.05) after 25.0 microg of deslorelin than after 12.5, 50.0 and 100.0 microg, which were not different from each other. Thus, no more than 12.5 microg of deslorelin is necessary to obtain maximum LH release in the model studied and doses less than 12.5 microg may also be effective.

The primary purpose of this experiment was to assess the possible beneficial effects of recombinant equine somatotropin (reST) administration on wound healing in adult geldings. The effects of the 21-d reST treatment on carbohydrate and lipid metabolism and on endogenous ST characteristics were monitored as well. Single, full-thickness skin incisions (7.62 x 7.62 cm) were made in the pectoral region of all geldings on d 0. Treated geldings received reST at 20 microg/kg BW i.m., and control geldings received vehicle (10 mM sodium borate) at equivalent volumes daily from d 0 (immediately after surgery) through d 20. Tracings of the wounds were made with acetate transparencies, and wound areas were calculated via a digital analyzer. In addition to once-daily blood samples collected at specified days throughout the treatment period, an i.v. glucose tolerance test was performed on d 16, and three assessments of endogenous ST secretion were performed in the 2 d immediately following the end of treatment: epinephrine administration during the morning of d 21, an exercise test during the afternoon of d 21, and i.v. aspartic acid infusion on d 22. There was no effect (P > . 1) of reST treatment on wound healing as assessed by changes in wound areas. Daily plasma ST, IGF-I, glucose, and insulin concentrations were higher (P < .05) and urea-nitrogen concentrations were lower (P < .001) in geldings receiving reST relative to controls. Glucose, NEFA, and insulin concentrations were all higher (P < .01) in reST-treated geldings before glucose infusion on d 16, and the responses to glucose were greater (P < .05) as well. Epinephrine administration increased (P < .02) ST concentrations in control geldings on d 21 but not in reST-treated geldings; a similar suppressive effect of reST treatment was observed for the ST response to exercise (P < .001). After aspartic acid infusion on d 22, reST-treated geldings had a much smaller (P < .001) ST response than did control geldings. In conclusion, reST administered to geldings at 20 microg/kg BW i.m. caused hyperglycemia, hyperinsulinemia, insulin insensitivity, mobilization of fatty acids, and an apparent negative feedback on the pituitary's ST response to various stimuli known to induce ST secretion. However, there was no beneficial effect of reST treatment with the wound model used in this experiment.

Thirty-two Suffolk wether lambs were fed for 84 d in a 2 x 2 factorial experiment using two levels of dietary protein (9.0 to 12.1% CP, low protein, LP; or 12.8 to 14.4% CP, high protein, HP) and supplemental Cr (none, C; or 400 ppb Cr as chromium tripicolinate, Cr). At 14- to 21-d intervals, lambs were weighed, and jugular blood samples were collected. Mean ADG and carcass weight (P > .10) did not differ. In lambs fed HP, Cr reduced liver weight and increased kidney weight (P < .01). Lambs fed HP had elevated plasma urea N (PUN; P < .01) and albumin (P < .04). During an i.v. epinephrine challenge on d 43, plasma cortisol declined in lambs fed Cr (Cr x time, P < .03) and in lambs fed LP (CP x time, P < .001). An i.v. glucose tolerance test conducted 3 h later showed that supplemental Cr decreased glucose clearance rate in lambs fed HP (CP x Cr, P < .10) but not in lambs fed LP. On d 62, PUN was increased in lambs fed HP (P < .001) between 0 and 3 h postprandial, and there was a Cr x CP interaction (P < .04). Postprandial plasma NEFA declined with Cr vs C (Cr x time, P < .07) and with HP vs LP (CP x time, P < .10). By d 66, lambs fed Cr had an elevated (P < .03) blood platelet and fibrinogen content. Chromium increased erythrocyte count in lambs fed HP (Cr x CP, P < .08), and isolated peripheral lymphocytes had greater blastogenic response to 4 microg/mL of phytohemagglutinin (Cr x CP, P < .001). The lymphocyte response to pokeweed mitogen (.2 microg/mL) was reduced in lambs fed Cr (P < .10). In the present experiment, Cr supplementation had minimal and inconsistent effects on production and metabolic criteria of lambs.

The effects of exogenous equine somatotropin (eST) administration on ovarian activity and plasma hormone levels were evaluated on horse and pony mares. The objectives of this study were to determine the effects of eST on follicular development and circulating concentrations of leutinizing hormone (LH), estradiol, progesterone, and insulin-like growth factor I (IGF-I) in cyclic horse and pony mares. Sixteen mares received daily injections (i.m.) of eST at a concentration of 25 micrograms/kg body weight on either Days 6 through 12 (Treatment A) or 13 through 19 (Treatment B) postovulation. In addition, contemporary mares were similarly given the carrier vehicle and served as controls (Treatments C and D). Blood samples were collected at 24-hr intervals and ultrasonographic evaluations were performed on the ovaries of each mare at 48-hr intervals beginning on the first day of treatment and ending either on the day of ovulation or 5 d postovulation. Circulating levels of insulin-like growth factor-I (IGF-I) were increased in treated mares by Day 3 post-treatment (P < 0.05). Also, mares in Treatment B exhibited a decrease in plasma estradiol concentrations (P < 0.05) when compared with control mares on Days 1 through 5 postovulation of the post-treated estrous cycle. In addition, circulating leutinizing hormone levels were different for mares in Treatment A compared with controls on Days--8 through--1 pre-ovulation (P < 0.05). All follicles present on the ovaries of each mare were measured and placed into one of five categories based on their diameter. Neither the mean number of follicles per size category > or = 8 mm in diameter nor the mean follicular diameter within each size category differed among treatment and control mares. However, eST treatment significantly increased the number of follicles < or = 7 mm on the ovaries of mares treated early in the estrous cycle when compared with control mares on Days 3 and 7 post-treatment and at the onset of standing estrus.

Forty-two Holstein calves were used to study performance and metabolic responses when milk replacer and then postweaning starter were supplemented with 1 ppm of Cr as Cr-tripicolinate. From birth through 8 wk of age, supplemental Cr tended to improve the growth performance of bull calves but not of heifer calves. Starter intake and feed efficiency were not affected by supplemental Cr. From 1 to 5 wk of age, plasma cortisol concentrations sampled just prior to feeding decreased, and concentrations of insulin-like growth factor-I increased. All calves appeared to become less sensitive to insulin as they aged. From 1 to 5 wk of age, plasma glucose and insulin concentrations gradually diverged for all calves; glucose concentrations decreased, and insulin concentrations increased. In addition, glucose clearance rate, measured by i.v. glucose tolerance tests, was more rapid when calves were 2 wk of age than when calves were 8 wk of age. The glucose clearance rate was greater in heifer calves than in bull calves but was not affected by supplemental Cr. Entry of plasma glucose following an i.v. propionate load was also greater in heifer calves than in bull calves but was not affected by supplemental Cr. Plasma nonesterified fatty acids were lower in calves fed milk replacer or starter supplemented with Cr than in control calves, although this effect diminished as calves aged. This finding was considered to be indirect evidence of enhanced insulin sensitivity in calves fed milk replacer or starter supplemented with Cr. Overall, data suggested that supplemental Cr-tripicolinate had minor effects on the metabolism and growth performance of conventionally managed dairy calves. The most notable effects occurred during the initial few weeks of life.

Gonadotropin-specific primary antisera and gold-conjugated secondary antibodies were used to immunocytochemically localize gonadotropins in the anterior pituitary of intact pony mares. Electron microscopy was then used to characterize the ultrastructure and immunoreactive staining characteristics of equine gonadotropes. Cells containing LH were morphologically indistinguishable from those containing FSH. Gonadotropes were relatively large and commonly had eccentric nuclei. The rough endoplasmic reticulum was well developed and dilated. Secretory granules were present in two morphologically distinct forms. Large polymorphic granules were generally located in perinuclear cytoplasmic areas, whereas small and uniformly shaped granules were in the peripheral cytoplasm, close to the cell membrane. Double-labeling revealed cells with granules that stained for both LH and FSH as well as cells that stained for either LH or FSH. Gonadotropes constituted 15 to 32% of all pituitary cells in the anterior pituitaries from the three mares included in this study. Cells that stained for only LH constituted 2 to 16% of all pituitary cells, cells that stained for only FSH ranged from 1 to 4.5%, and cells staining for both hormones constituted 6.2 to 24% of the pituitary cells. These results indicate that there are in fact three distinct subclasses of gonadotropes in the equine anterior pituitary based on immunocytochemical staining, which is similar to the situation described for several other mammalian species.

When aspirating ovarian follicles in pregnant mares to obtain oocytes for in vitro fertilisation (IVF), the effect of the manipulation on circulating concentrations of progesterone may be an important consideration in terms of the maintenance of pregnancy. The object of this study was to compare the effects of 3 different forms of transvaginal ultrasound-guided follicle aspiration (Treatment 1, no aspiration, n = 4; Treatment 2, aspirate only follicles > or =20 mm in diameter, n = 7; Treatment 3, aspirate all visible follicles, n = 7) on peripheral plasma progesterone concentrations between Days 21 and 150 of gestation in 9 mares carrying intraspecies horse and 9 mares carrying interspecies mule conceptuses. The 3 follicle aspiration treatments were applied at the peak of each follicular wave as determined by follicular mapping by means of transrectal ultrasonography on alternate days. The plasma progesterone profile in mares undergoing Treatment 1 was in close agreement with those reported previously in pregnant mares. A decline in plasma progesterone levels occurred after Day 53 of gestation in Treatments 2 and 3 mares, indicating that the follicular aspiration procedures did interfere with the formation of secondary corpora lutea. However, the levels in individual mares never dropped low enough to endanger the pregnancy. Mares carrying mule pregnancies exhibited higher mean plasma progesterone concentrations between Days 39 and 45 of gestation than mares carrying horse pregnancies, equivalent levels between Days 46 and 66 despite the lower circulating concentrations of chorionic gonadotrophin (mule CG) in their blood during this period and lower progesterone levels between Days 67 and 150 of gestation. The results indicate that the primary corpus luteum in the pregnant mare may be more sensitive to mule CG than horse CG. Furthermore, the earlier disappearance of CG from the circulation in mares carrying mule fetuses is reflected by an earlier decline in plasma progesterone concentrations in this type of equine pregnancy.

The ultrastructural and immunoreactive staining characteristics of cells containing prolactin (lactotropes) and growth hormone (GH; somatotropes) in the anterior pituitaries of gonadally intact pony mares were studied at the electron microscopic level. Lactotropes included two morphological subsets: Type I cells were larger and contained large, dense, polymorphic granules that were scattered throughout the cytoplasm; Type II cells were smaller and contained small, dense, polymorphic granules that were predominantly found in peripheral areas of the cytoplasm. Lactotropes constituted 5 to 16% of the total number of cells in the pituitary. Somatotropes were medium-sized cells containing uniform, large, dense secretory granules. The somatotropes contained the largest secretory granules in the pituitary and represented 11 to 26% of the total number of cells. Type I lactotropes and somatotropes were readily distinguishable without immunocytochemical staining. Double-labeling of pituitary sections allowed for characterization of cells that contained both hormones (mammosomatotropes). These cells were morphologically indistinguishable from Type I lactotropes and constituted 6.5 to 16.5% of the total number of cells. Results from this study demonstrated that there are two cell populations that contain only prolactin (Type I and II lactotropes) and one cell population that contains only GH (somatotropes) in the equine pituitary, and an additional subset of cells that contains GH and prolactin in the same secretory granules.

This experiment was designed to determine 1) the efficacy of daily s.c. injections of a dopamine antagonist, sulpiride, for increasing prolactin secretion in geldings in winter and 2) whether increasing prolactin concentrations would hasten the onset of hair shedding or enhance gonadotropin secretion. Five geldings each received vehicle (vegetable oil) or sulpiride (100 mg in vehicle) daily from February 8 through March 29. On February 8 and every 7 d thereafter through March 29, blood samples were drawn around treatment injections and hair samples were collected. On March 30, all geldings received an injection of GnRH and thyrotropin-releasing hormone. Over the 8-wk sampling period, prolactin response to sulpiride varied in a quadratic manner (P < .002). Average area under the 2-h response curve for sulpiride-treated geldings was 24.9 h.ng.mL-1 on February 8, declined to 4.3 after 4 wk, and then increased to 14.8 by the 8th wk of treatment (pooled SE = 4.1 h.ng.mL-1). Prolactin concentrations in control geldings did not vary (P > .1) after injection or over the 8-wk period. Weight of hair pulled from sulpiride-treated geldings did not peak as sharply or as high as that from control gelding (P < .05) and continued to be high through the 11th wk. Sulpiride treatment reduced (P = .071) the LH response to GnRH on March 30; the FSH and prolactin responses to secretagogue were not altered (P > .25). In conclusion, even though prolactin concentrations were increased by sulpiride, the effects on gonadotropin secretion and hair shedding were minor and opposite of those expected.

Eight pony mares received 4 mg of recombinant porcine prolactin (rpPRL) daily for 45 d beginning on January 15; eight control mares received vehicle. Reproductive end points and various indicators of metabolism, hair shedding, and thyroid activity were monitored. Prolactin concentrations peaked in mares treated with rpPRL at 94 +/- 19 ng/mL 2 h after injection and were 5.1 +/- 1.7 ng/mL 24 h after injection. Treatment with rpPRL increased (P < .01) hair shedding within 14 d, which peaked at 28 d and then dropped precipitously. Binding of 125I-equine prolactin confirmed that antibodies were present in rpPRL-treated mares by 28 d, thus only data up to that time are included herein. The percentage of mares in estrus after 17 d of treatment was greater (P < .05) for mares treated with rpPRL than for controls, although the average day of estrus onset did not differ (P > .1). The date of first ovulation, determined from plasma progesterone concentrations, was hastened (P < .005) in mares treated with rpPRL (February 6 +/- 3 d) relative to control mares (March 14 +/- 6 d). Concentrations of insulin-like growth factor I and nonesterified fatty acids were not affected (P < .05) over time. There were minor differences between groups in thyroid hormone secretion. Various assessments of glucose and insulin indicated no perturbation (P > .1) due to treatment with rpPRL. We conclude that prolactin mediates the onset of vernal hair shedding in mares and is in some way involved with ovulation.

Two experiments were conducted to determine 1) the prolactin response to different kinds of feedstuffs in stallions and 2) the effects of total feed deprivation on prolactin secretion in mares and its interaction with the prolactin response to feeding. Experiment 1 was performed with stallions as a 6 x 6 Latin square: A) no feed; B) pelleted feed fed to meet 82.5% of the horses' CP requirements; C) pelleted feed at 25% of the amount in B; D) pelleted feed as in B plus water ad libitum; E) cracked corn at the weight in B; and F) chopped alfalfa at the weight in B. The positive prolactin responses (P < .05) to feeding were similar for treatments B through F. The insulin response to feeding was highest (P < .05) in stallions fed water with the pelleted feed. In Exp. 2, 72 h of feed deprivation did not affect (P > .1) daily prolactin secretion. Feeding of a meal on the 3rd d of deprivation increased (P < .05) plasma prolactin, insulin, and glucose concentrations similarly in all mares. There was a positive growth hormone response (P < .1) after feeding in feed-deprived mares but not in fed mares. The prolactin response (P < .001) to thyrotropin-releasing hormone was greater (P = .083) for feed-deprived mares than for controls, whereas the response to sulpiride (P < .001) only tended to differ (P = .16) between groups. We conclude that prolactin secretion may be stimulated by aspects of eating other than the feedstuff itself. Total feed deprivation had little effect on the subsequent prolactin response to a meal or to other known secretagogues.

To test the hypothesis that prolactin mediates the increase in seminal volumes induced by sexual stimulation in stallions, semen was collected from six stallions every other day for 26 d. The last eight collection days were treatment days. For each stallion, four treatments were randomly assigned to the first four of the eight treatment collection days, and then repeated in reverse order on the last four collection days; 1) CONTROL: semen collected per normal procedures; 2) Sexually stimulated: stallions were presented to mares in a chute for 10 min before collection; 3) Bromocriptine (dopamine agonist) plus sexual stimulation: stallions were administered bromocriptine 10 min before 10 min of sexual stimulation prior to collection; and 4) Sulpiride: stallions were administered sulpiride (a dopamine antagonist) 25 min before collection. Prolactin concentrations in plasma were increased by sexual stimulation (P < .01) and sulpiride (P < .001) administration and were decreased (P < .01) when bromocriptine was administered before sexual stimulation. Sexual stimulation alone increased (P < .01) volume of gel-free semen relative to control values (102 vs 81 mL), and bromocriptine prevented this response (89 mL; P < .075 relative to sexual stimulation). Sulpiride had no effect (P > .1) on gel-free volume. Volume of gelatinous material, number of mounts, sperm concentration, motility, pH of gel-free semen, number of spermatozoa per ejaculate, and prolactin concentration in gel-free semen were not affected (P > .1) by treatment. Although the effect of bromocriptine indicated that prolactin may mediate the rise in seminal volume due to sexual stimulation, if it does, it must be in conjunction with other hormone(s) or factor(s), because increasing prolactin concentrations alone did not increase seminal volume in these stallions.

Two diets consisting of bermudagrass hay and a corn-cottonseed hull-based supplement were formulated to provide either 100% (control) or 50% (restricted) of the protein and energy requirements for maintenance for mature mares. Twelve light horse mares were fed the control diet for 7 d, and then at 0800 on d 0, six mares were switched to the restricted diet. All diets were fed as two equally sized meals at 0800 and 1600. At 0800 on d 7, mares receiving the restricted diet were switched back to the control diet. Relative to control mares, mares switched to the restricted diet had reduced plasma concentrations of glucose (P = .005) and insulin (P = .09) in response to the two restricted meals on d 0. However, concentrations of both glucose and insulin returned to control levels (P > .1) within 1 h after the consumption of the control diet on d 7. Dietary restriction increased (P = .009) plasma NEFA concentrations within the first 24 h, and NEFA concentrations remained elevated (P < .001) in restricted mares until the mares were returned to the control diet on d 7. Meal-induced increases (P < .05) in plasma concentrations of glucose, insulin, urea N, glucagon, and thyroxine were observed. These results emphasize the importance of early sampling when monitoring plasma constituents during nutrient alterations and indicate that prefeeding responses of plasma constituents alone may not fully explain the metabolic consequences of nutrient restriction.

A combination regimen of equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) was used to stimulate ovarian follicular development in domestic cats. The rate of elimination of eCG from circulation was estimated, and, following follicular aspiration, the formation of ancillary follicles and secondary CL was characterized. The effect of gonadotropin-neutralizing antisera on the development of secondary ovarian structures, CL function and humoral immune responses also was evaluated. After intramuscular injection, initial serum eCG concentrations were variable, with the elimination half-life estimated at 39 to 55 h and eCG persisting in circulation for several days. Following follicular aspiration, queens formed CL equal to the number of aspirated follicles and exhibited a rapid increase in progesterone concentration but developed high numbers of ancillary follicles by 5 d post aspiration. By 15 d post aspiration, all ancillary follicles had luteinized to form secondary CL. Treatment with neutralizing antisera at the time of follicular aspiration slowed (P < 0.05) CL formation but did not decrease (P > 0.05) the number of ancillary follicles or secondary CL. Progesterone concentrations did not differ (P > 0.05) from control queens while secondary humoral immune responses to eCG were qualitatively similar between groups. In summary, eCG was eliminated slowly from cats following intramuscular injection and this persistence in circulation may have contributed to the development of ancillary follicles and secondary CL. However, the administration of neutralizing antisera at the time of follicular aspiration was ineffective in preventing the formation of these secondary ovarian structures.

Twelve light horse mares were fed a control diet that provided 100% of their maintenance protein and energy requirements for 7 d and were then either continued on the control diet or totally deprived of feed (with access to water) for 3 d . Plasma samples were drawn twice daily throughout the experiment, at 15-min intervals for 9 h beginning 45 h after feed removal, and at 10-min intervals around an exercise bout beginning 73 h after feed removal. Feed deprivation increased (P < or = .06) whole blood beta-hydroxybutyrate and plasma NEFA, urea N, L-lactate, and glucagon concentrations, decreased (P = .02) IGF-I concentrations, and did not change (P > .1) plasma glucose insulin, prolactin, triiodothyronine, and thyroxine concentrations. Exercise increased (P < .05) plasma NEFA, prolactin, and growth hormone (GH) concentrations in all mares. Plasma NEFA concentrations increased (P < .001) after exercise and remained increased in fed mares, but rapidly decreased in deprived mares (time x diet interaction, P = .006). Plasma glucose concentrations following exercise increased in deprived mares but decreased in fed mares (time x diet interaction, P = .07). The plasma prolactin response after exercise also differed between groups (P = .09). Feed-deprived mares had greater (P = .02) plasma GH concentrations before exercise (73 h after feed withdrawal) and had a greater (P < .001) GH peak at 10 min after initiation of exercise. The increase in secretion rate o GH due to feed deprivation in these mares was similar to that reported for other domestic species but was not nearly as great in magnitude.

The influence of slaframine (SF), a parasympathomimetic compound isolated from the fungus Rizoctonia leguminicola, on circulating metabolic hormone concentrations was investigated in goats. In Exp. 1, SF was administered i.v. at 0 (CONT), 50 (LSF), 100 (MSF), or 150 (HSF) microgram/kg.75 BW in four mature Spanish-cross does (average BW 36 +/- 7 kg) fitted with indwelling jugular vein catheters in a 4 x 4 Latin square design. Plasma glucose peaked (P < .06) at 120 min with LSF and at 180 min with HSF and was higher (P <.06) than the CONT at these times. Glucose exhibited a quadratic response (P < .03) to SF. Area under the response curve for glucose differed (P < .02) in HSF from CONT and MSF. Insulin peaked (P < .01) at 240 min with MSF and at 180 min with HSF. Plasma triiodothyronine was maintained at a higher level (P < .03) with HSF. Thyroxine peaked (P < .06) at 120 min with MSF and 300 min with HSF. Plasma NEFA and somatotropin concentrations were not affected (P > .10) by SF. In Exp. 2, four mature Spanish-cross wethers (average BW 27 +/- 2 kg) fitted with jugular vein catheters were administered SF (0 and 114 micrograms/kg.75 BW) and 4-diphenylacetoxy-N-methylpiperidine methiodide (4DAMP; 0 and 258 micrograms/kg.75 BW), a M3-muscarinic receptor antagonist, i.v. in a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. With SF, glucose peaked (P < .06) at 60 min and insulin peaked (P < .05) at 180 min. Plasma triiodothyronine levels were maintained (P < .05) with SF but declined with other treatments. Plasma NEFA and thyroxine concentrations remained unchanged regardless of treatment. Slaframine administration induced hyperglycemia and hyperinsulinemia in goats; however, these changes were blocked by preadministration of isomolar quantities of the M3-muscarinic receptor antagonist, 4DAMP.

Gross observation of testicular parenchyma of 1.5- to 2-yr-old horses reveals both light and dark regions. If this gross, differential shading reflects quantitative differences in the development of spermatogenesis and interstitial cell populations, the horse may prove to be a useful model for study of the paracrine relationships associated with initiation of spermatogenesis. The objective of this study was to characterize seminiferous tubules and interstitium of testes with gross, differential shading. Testes with both light and dark regions of parenchyma were obtained from horses 1.5-2 yr old and compared to parenchyma of fetal, 2-yr-old, or 5-yr-old horses. Stereology was used on tubular and interstitial components, and luminal development of seminiferous tubules was scored. Volume density of seminiferous tubules, percentage of tubules with large vacuoles or a complete lumen, and number of primary spermatocytes per gram were greater (p < 0.05) in light parenchyma than in dark parenchyma. The percentage of tubules with no lumen and the percentage of parenchyma occupied by interstitial space were greater (p < 0.05) in fetal and dark parenchyma than in light parenchyma. The number of Leydig cells per gram parenchyma was similar (p > 0.05) in both light parenchyma and dark parenchyma. A greater percentage (p < 0.05) of other (nonvascular, non-Leydig, nonmacrophage) cells was found in the dark parenchyma than in light parenchyma or in testes of 2- or 5-yr-old horses. The volume density of macrophages was notably greater (p < 0.05) in fetal and dark parenchyma than in light parenchyma or in testes from older horses. Variation in development of seminiferous tubules was not associated with the volume density of blood vessels. In conclusion, the gross, differential shading of equine testicular parenchyma with its corresponding differences in seminiferous tubular development is a clear example of the effect of local factors leading to the local initiation of spermatogenesis.