Oral and intravenous carbohydrate challenges decrease active ghrelin concentrations and alter hormones related to control of energy metabolism in horses

This study tested the hypothesis that grain and intravenous dextrose challenges would alter plasma concentrations of active ghrelin, adiponectin, leptin, glucose, insulin, and cortisol in Standardbred mares. To deliver 0.5 g of glucose (dextrose solution for the intravenous test)/kg of BW, mares received intravenous dextrose (50% solution) or oral grain administration in 2 trials. In response to the oral grain challenge, plasma glucose and insulin concentrations increased (P < 0.001) by 56 and 802%, respectively. Plasma ghrelin concentration initially decreased (P < 0.001) by 40%, then subsequently increased (P < 0.05) from its nadir by 259%. Plasma leptin concentration decreased (P = 0.002) 17% compared with baseline. There was no change (P = 0.34) in plasma adiponectin concentration in response to oral grain challenge; however, plasma cortisol concentrations decreased (P < 0.001) by 24%. In response to the intravenous dextrose challenge, plasma glucose and insulin concentrations increased (P < 0.001) by 432 and 395%, respectively. Plasma active ghrelin concentration initially decreased (P < 0.001) by 56%, then subsequently increased (P < 0.001) from its nadir by 314%. Plasma leptin concentration also increased (P < 0.001) by 33% compared with baseline. There was no change (P = 0.18) in plasma adiponectin concentration throughout the dextrose challenge. Plasma cortisol concentration increased (P = 0.027) by 20%. Hence, oral grain and intravenous nutrient challenges have the ability to alter variables potentially related to energy metabolism in mares, with acute changes in glucose and insulin possibly modulating changes in ghrelin and leptin.     

Endothelin response during and after exercise in horses

The objective of the present study was to measure plasma endothelin-1 (ET-1) at rest and during exercise in the horse. Six healthy, Standardbred and Thoroughbred mares (5.3+/-0.8 years; 445.2+/-13.1 kg) which were unfit, but otherwise accustomed to running on the treadmill, were used in the study. Plasma ET-1 concentrations were measured using a commercially available radioimmunoassay kit. Horses performed three trials: a standing control (CON) trial where blood was collected from the jugular vein every minute for 5 min; a graded exercise test (GXT) where blood samples were collected at the end of each 1 min step of an incremental exercise test; and a 15 min submaximal (60% VO(2max)) steady-state exercise test (SST) where blood samples were collected 1 min before, immediately after, and at 2 min, 10 min and 20 min post-exercise. Plasma ET-1 concentration did not change (P>0.05) during the CON trial where it averaged 0.18+/- 0.03 pg/mL (mean+/-SE). Surprisingly, plasma ET-1 concentration did not change during the GXT trial where it averaged 0.20+/-0.03 pg/mL. There were no differences between the mean concentrations obtained in either trial (P>0.05). Plasma ET-1 concentrations were, however, significantly elevated (P<0.05) immediately following exercise and at 2 min post-exercise in the SST. Post-exercise plasma ET-1 concentrations returned to baseline (P>0.05) by 10 min of recovery. Together, these data may suggest that ET-1 concentrations are altered in response to an exercise challenge.     

Diurnal variation of ghrelin, leptin, and adiponectin in Standardbred mares

Twelve Standardbred mares underwent blood sampling for 24 h to test the hypothesis that there is diurnal variation of humoral mediators of peripheral energy balance including active ghrelin, adiponectin, leptin, glucose, insulin, and cortisol. The experiment was conducted under acclimated conditions. Grass hay and pelleted grain were provided at 0730 and 1530. Plasma concentrations of active ghrelin and leptin concentrations both peaked (47.3 +/- 6.5 pg/ mL and 5.9 +/- 1.1 ng/mL, respectively; P < 0.05) at 1550, 20 min after feeding. Active ghrelin decreased (P < 0.05) to 28.9 +/- 4.5 pg/mL overnight. The nadir of leptin (4.6 +/- 0.9 ng/mL) occurred at 0650. Neither hormone showed variation (P > 0.05) after the morning feeding. Plasma glucose and insulin concentrations increased (P < 0.05) in response to feeding; however, the morning responses (glucose = 96.9 +/- 2.6 mg/dL; insulin = 40.6 +/- 7.3 uIU/mL) were greater (P < 0.05) than the afternoon responses (glucose = 89.9 +/- 1.8 mg/dL; insulin = 23.2 +/- 4.3 uIU/mL at 180 and 60 min after feeding, respectively). Cortisol concentrations increased (P < 0.05) during the morning hours, but did not respond to feeding, whereas adiponectin concentrations remained stable throughout the study. Hence, active ghrelin and leptin may be entrained to meal feeding in horses, whereas adiponectin seems unaffected. We concluded that there seems to be a diurnal variation in glucose and insulin response to a meal in horses. Furthermore, elevated glucose and insulin concentrations resulting from the morning feeding may be responsible for the increase in leptin concentration in the afternoon.     

Effects of fasting and refeeding on plasma concentrations of leptin, ghrelin, insulin, growth hormone and metabolites in swine

The effects of nutritional status, such as fasting and refeeding, on leptin and ghrelin secretion in swine were examined. The swine (n = 4) were fasted for 54 h and plasma hormone levels were measured before, during and after fasting. Plasma leptin and insulin concentrations began to decrease 12 and 6 h into the fasting period, respectively (P < 0.05), and maintained a low level for the remaining period of fasting. Plasma leptin and insulin returned to the pre‐fasting value 6 and 12 h after refeeding, respectively. Plasma ghrelin concentrations showed a nocturnal periodicity during the fasting period; it increased nocturnally at 36 and 42 h into the fasting period (P < 0.05). Plasma growth hormone levels did not show any remarkable changes during the fasting. Plasma glucose levels showed a modest fall during fasting and significantly decreased (P < 0.05) at 24 h into the fasting period, returning to pre‐fasting levels after refeeding. Plasma nonesterified fatty acid levels increased (P < 0.05) at 12 h into the fasting period and returned to the pre‐fasting level 6 h after refeeding. These results indicate that plasma leptin, insulin and ghrelin play an important role in maintaining energy homeostasis in swine. The plasma ghrelin did not continuously increase, but showed nocturnal periodicity during fasting. This may suggest that ghrelin is also involved in physiological processes other than energy homeostasis.     

Effects of amino acids infused into the vein on ghrelin‐induced GH,insulin and glucagon secretion in lactating cows

To investigate the effects of amino acids on ghrelin‐induced growth hormone (GH), insulin and glucagon secretion in lactating dairy cattle, six Holstein cows were randomly assigned to two infusion treatments in a cross‐over design. Mixture solution of amino acids (AMI) or saline (CON) was continuously infused into the left side jugular vein via catheter for 4 h. At 2 h after the start of infusion, synthetic bovine ghrelin was single injected into the right side jugular vein through the catheter. Ghrelin injection immediately increased plasma GH, glucose and non‐esterified fatty acids (P < 0.05) with no difference between both treatments. Additionally, plasma insulin and glucagon concentrations were increased by ghrelin injection in both treatments. The peak value of plasma insulin concentration was greater in AMI compared with CON (P < 0.05). Plasma glucagon concentration showed no difference in the peak value reached at 5 min between both treatments, and then the plasma levels in AMI compared with CON showed sustained higher values (P < 0.05). After plasma glucose concentration reached the peak, the decline was greater in AMI compared with CON (P < 0.05). These results showed that the increased plasma amino acids may enhance ghrelin action which in turn enhances insulin and glucagon secretions in lactating cows.     

Compensation for obesity-induced insulin resistance in dogs: assessment of the effects of leptin, adiponectin, and glucagon-like peptide-1 using path analysis

The hormonal mediators of obesity-induced insulin resistance and compensatory hyperinsulinemia in dogs have not been identified. Plasma samples were obtained after a 24-h fast from 104 client-owned lean, overweight, and obese dogs. Plasma glucose and insulin concentrations were used to calculate insulin sensitivity and β-cell function with the use of the homeostasis model assessment (HOMA_{insulin sensitivity} and HOMA_{β-cell function}, respectively). Path analysis with multivariable linear regression was used to identify whether fasting plasma leptin, adiponectin, or glucagon-like peptide-1 concentrations were associated with adiposity, insulin sensitivity, and basal insulin secretion. None of the dogs were hyperglycemic. In the final path model, adiposity was positively associated with leptin (P < 0.01) and glucagon-like peptide-1 (P = 0.04) concentrations. No significant total effect of adiposity on adiponectin in dogs (P = 0.24) was observed. If there is a direct effect of leptin on adiponectin, then our results indicate that this is a positive relationship, which at least partly counters a negative direct relationship between adiposity and adiponectin. Fasting plasma leptin concentration was directly negatively associated with fasting insulin sensitivity (P = 0.01) and positively associated with β-cell function (P < 0.01), but no direct association was observed between adiponectin concentration and either insulin sensitivity or β-cell function (P = 0.42 and 0.11, respectively). We conclude that dogs compensate effectively for obesity-induced insulin resistance. Fasting plasma leptin concentrations appear to be associated with obesity-associated changes in insulin sensitivity and compensatory hyperinsulinemia in naturally occurring obese dogs. Adiponectin does not appear to be involved in the pathophysiology of obesity-associated changes in insulin sensitivity.     

Effects of cool and hot humid environmental conditions on neuroendocrine responses of horses to treadmill exercise

To determine the effects of exercise, high heat and humidity and acclimation on plasma adrenaline, noradrenaline, beta-endorphin and cortisol concentrations, five horses performed a competition exercise test (CET; designed to simulate the speed and endurance test of a three-day event) in cool dry (CD) (20 degrees C/40% RH) and hot humid (30 degrees C/80% RH) conditions before (pre-acclimation) and after (post-acclimation) a 15 day period of humid heat acclimation. Plasma adrenaline and noradrenaline concentrations pre-acclimation were significantly increased compared with exercise in the CD trial at the end of Phases C (P<0.05) and D (P<0.05 and P<0.01, respectively) and at 2 min recovery (P<0.01), with adrenaline concentrations still elevated after 5 min of recovery (P<0.001). Plasma beta-endorphin concentrations were increased at the end of Phases C (P<0.05) and X (P<0.01) and at 5 and 30 min recovery (P<0.05) in the pre-acclimation session. Plasma cortisol concentrations were elevated after the initial warm up period pre-acclimation (P<0.01) and at the end of Phase C (P<0.05), compared with the CD trial. A 15 day period of acclimation significantly increased plasma adrenaline concentrations at 2 min recovery (P<0.001) and plasma cortisol concentration at the end of Phase B (P<0.01) compared with pre-acclimation. Acclimation did not significantly influence noradrenaline or beta-endorphin responses to exercise, although there was a trend for plasma beta-endorphin to be lower at the end of Phases C and X and after 30 min recovery compared with pre-acclimation. Plasma adrenaline, noradrenaline, beta-endorphin and cortisol concentrations were increased by exercise in cool dry conditions and were further increased by the same exercise in hot humid conditions. Exercise responses post-acclimation suggest that adrenaline and noradrenaline may play a role in the adaptation of horses to thermal stress and that changes in plasma beta-endorphin concentrations could be used as a sensitive indicator of thermal tolerance before and after acclimation. The use of plasma cortisol as a specific indicator of heat stress and thermal tolerance before or after acclimation in exercising horses appears limited.     

Chronic clenbuterol administration alters myosin heavy chain composition in standardbred mares

The purpose of this study was to examine changes in myosin heavy chain (MHC) composition due to chronic clenbuterol administration with or without exercise in mares. Unfit Standardbred mares (aged 10+/-3 years) were divided into four groups: clenbuterol (2.4 micro/kg BW twice daily) plus exercise (3 days/week for 20 min at 50% VO(2max); CLENEX; n=6), clenbuterol only (CLEN; n=6), exercise only (EX; n=5), and control (CON; n=6). Muscle biopsies were obtained from gluteus medius muscle before and after the eight-week training/administration period. MHC composition was determined via SDS gel electrophoresis and quantified using a scanning and densometric system.CLENEX and CLEN exhibited significant (P<0.05) MHC changes while EX and CON did not. MHC type IIA decreased (29.8+/-6.1 to 19.3+/-4.0%, CLENEX; and 36.8+/-12.4 to 26.4+/-7.9%, CLEN) and MHC type IIX increased (59.4+/-7.2 to 71.8+/-5.8%, CLENEX; and 50.5+/-12.5 to 62.0+/-9.3%, CLEN). Chronic clenbuterol administration with and without exercise resulted in a significant shift in MHC profile in Standardbred mares.     

High versus low body condition in mares: interactions with responses to somatotropin,GnRH analog,and dexamethasone

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 effect of interval versus continuous exercise on plasma leptin and ghrelin concentration in young trotters

The effect of interval vs. continuous exercise on plasma leptin and ghrelin concentration in young Standardbred horses was studied. The experiment was conducted on 27 trotters, in the age between 2 and 3 years. They were divided into two groups according to the type of exercise. Blood samples were collected through jugular venipuncture in the following experimental conditions: at rest, immediately after exercise and 30 minutes after the end of the effort. Plasma leptin and ghrelin concentrations were determined using RIA tests. The continuous exercise induced an increase in plasma leptin concentration whereas the interval type of exercise did not influence the level of this hormone (3.47 +/- 0.78 vs. 4.07 +/- 0.94 and 2.31 +/- 0.15 vs. 2.36 +/- 0.21 ng/mL, respectively). The plasma ghrelin concentration measured after the continuous exercise, significantly increased (720 +/- 27.4 vs. 814 +/- 13.8; p < or = 0.05) whereas concentration of this hormone assessed after the interval exercise, significantly dropped (982 +/- 56.5 vs. 842 +/- 35.6 pg/mL; p < or = 0.05). The changes in plasma ghrelin concentration measured after the end of the effort correlated inversely with blood lactic acid concentration. In conclusion, the obtained results showed that medium-intensive type of exercise, such as trot, interval or continuous, slightly affected plasma leptin level but significantly affected plasma ghrelin concentration in young Standardbred trotters.     

Propionate is not an important regulator of plasma leptin concentration in dairy cattle

Propionate was recently shown to increase leptin synthesis in rodents. To determine if a similar effect occurs in ruminants, propionate was administered to lactating dairy cows. In experiment 1, 31 cows were given an intrajugular Na propionate bolus (1,040 micromol/kg body weight), increasing plasma propionate from 160 to 5,680 microM and plasma insulin from 6.8 to 77.8 microIU/mL. Plasma leptin concentration decreased from 2.11 ng/mL before bolus to 1.99 ng/mL after dosing (P<0.05) with no differences in leptin concentrations at 20, 50, and 100 min post-bolus (P>0.10). In experiment 2, 12 cows were used in a duplicated 6 x 6 Latin square experiment to assess the dose-response effect of ruminal propionate infusion on plasma leptin concentration. Sodium propionate was infused at rates of 0, 260, 520, 780, 1040, or 1,300 mmol/h, while total short-chain fatty acid infusion rate was held constant at 1,300 mmol/h by addition of Na acetate to the infusate. Coccygeal blood was sampled following 18 h of infusion. Increasing the rate of propionate infusion linearly increased plasma propionate concentration from 180 to 330 microM (P<0.001) and plasma insulin concentration from 6.7 to 9.1 microIU/mL (P<0.05). There was a quadratic response in plasma leptin concentration (P=0.04) with a maximum at 780 mmol/h propionate, but leptin concentrations increased by no more than 8% relative to the 0 mmol/h propionate infusion. Leptin concentrations were correlated with insulin concentrations but not with propionate concentrations in plasma. Propionate is not a physiological regulator of leptin secretion in lactating dairy cows.     

Effects of lactation on metabolic and reproductive hormones in Lipizzaner mares

In this study, growth hormone (GH), insulin-like growth factor 1 (IGF-1), leptin, luteinising hormone (LH) and prolactin were analyzed in mares from late pregnancy throughout lactation (group 1, n=46) and in non-lactating mares (group 2, n=11). Plasma GH concentrations in group 1 mares during gestation and lactation were lower than in mares of group 2 (P<0.05). Highest IGF-1 levels were found in lactating mares in the week of foaling. IGF-1 concentrations decreased continuously thereafter. Plasma leptin concentrations decreased after foaling and, for 4 weeks, were lower in lactating than in non-lactating mares (P<0.05). Reduced leptin concentrations may promote feed intake and allow lactating mares to avoid an energy deficit. In group 1 mares, prolactin concentrations reached a maximum in the week of foaling and decreased rapidly thereafter. Plasma LH concentrations in group 1 mares before foaling were lower than at corresponding times in group 2 (P<0.05). LH concentrations then increased and did no longer differ from group 2 until week 2 postpartum. This increase may contribute to the resumption of cyclic ovarian activity in postpartum mares. Subsequently, LH levels in lactating mares decreased again (P<0.05). Increased IGF-1 concentrations early postpartum might contribute to ovarian stimulation while reduced IGF-1 and GH concentrations later in lactation might cause reduced stimulation. The changes in somatotrophic hormones could thus explain, at least in part, a more pronounced stimulation of ovarian function early postpartum than during the following months of lactation.     

Leptin and Ghrelin and the Indices of Lipid Metabolism as Related to Sex Steroid Hormones in Trotters

This study examined the influence of sex steroid hormones on lipid metabolism in horses. The group of 34 clinically healthy Standardbred trotters aged 2 to 4 years was studied during an exercise test. The horses were divided into groups according to their sex. These groups were: 11 stallions, 16 mares, and seven geldings. Concentrations of testosterone, 17-β-estradiol, leptin, ghrelin, glycerol, free fatty acids (FFA), and triacylglycerols (TG) were measured in plasma obtained from blood samples taken at rest and after the end of the exercise. At rest, plasma ghrelin concentration was significantly higher in geldings than in stallions and mares (1,541 ± 206 vs 1,280 ± 288 and 1,310 ± 267 pg/mL, respectively; P = .012). Leptin was lower in geldings than in mares (2.65 ± 0.93 vs 4.70 ± 2.31 ng/mL; P = .036). The post-exercise rise in plasma ghrelin and TG concentrations was significantly higher in mares than in geldings (+220 ± 330 vs -25 ± 206 pg/mL; P = .049 and 0.31 ± 0.14 vs 0.13 ± 0.15 mmol/L; P = .016, respectively). The increase in plasma FFA level was higher in geldings than in stallions (535 ± 178 vs 334 ± 191 μmol/L, P = .046). In conclusion, lipolysis rate in geldings is higher than in noncastrated trotters.     

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

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

Effects of short-term feed deprivation and melatonin implants on circadian patterns of leptin in the horse

Leptin is a protein hormone produced by adipose tissue that influences hypothalamic mechanisms regulating appetite and energy balance. In species tested thus far, including horses, concentrations of leptin increase as animal fat mass increases. The variables and mechanisms that influence the secretion of leptin are not well known, nor is it known in equine species how the secretion of leptin is influenced by acute alterations in energy balance, circadian patterns, and/or reproductive competence. Our objectives were to determine in horses: 1) whether plasma concentrations of leptin are secreted in a circadian and/or a pulsatile pattern; 2) whether a 48-h period of feed restriction would alter plasma concentrations of leptin, growth hormone, or insulin; and 3) whether ovariectomy and/or a melatonin implant would affect leptin. In Exp. 1, mares exposed to ambient photoperiod of visible light (11 h, 33 min to 11 h, 38 min), received treatments consisting of a 48-h feed restriction (RES) or 48 h of alfalfa hay fed ad libitum (FED). Mares were maintained in a dry lot before sampling and were tethered to a rail during sampling. Analyses revealed that leptin was not secreted in a pulsatile manner, and that mean leptin concentrations were greater (P < 0.001) in FED vs. RES mares (17.20 +/- 0.41 vs. 7.29 +/- 0.41 ng/mL). Plasma growth hormone was pulsatile, and mean concentrations were greater in RES than FED mares (2.15 +/- 0.31 vs. 1.08 +/- 0.31 ng/mL; P = 0.05). Circadian patterns of leptin secretion were observed, but only in FED mares (15.39 +/- 0.58 ng/mL for morning vs. 19.00 +/- 0.58 ng/mL for evening; P < 0.001). In Exp. 2, mares that were ovariectomized or intact received either a s.c. melatonin implant or a sham implant. Thereafter, blood was sampled at weekly intervals at 1000 and 1700. Concentrations of leptin in samples collected at 1700 were greater (P < 0.001) than in those collected at 1000 (28.24 +/- 1.7 vs. 22.07 +/- 1.7 ng/mL). Neither ovariectomy nor chronic treatment with melatonin affected plasma concentrations of leptin or the circadian pattern of secretion. These data provide evidence that plasma leptin concentrations in the equine are sensitive to acute changes in nutritional status and vary in a circadian pattern that is sensitive to fasting but not to melatonin treatment or ovariectomy.     

Seasonal changes in the interactions among leptin, ghrelin, and orexin in sheep

The adaptation of the physiology of an animal to changing conditions of light and food availability is evident at the behavioral and hormonal levels. Melatonin, leptin, ghrelin, and orexin, which exhibit rhythmic secretion profiles under ad libitum feeding conditions, are sensitive to changes in daylength, forming a tight web of interrelationships in the regulation of energy balance. The aim of this study was to determine the effects of central injections of leptin, ghrelin, and orexin on the reciprocal interactions among these hormones and the influence of photoperiod on these responses. Twenty-four ovariectomized and estradiol-implanted ewes were used in a replicated switchback design. The ewes were assigned randomly to 1 of 6 treatment groups, and the treatments were infused into their third ventricles 3 times at 0, 1, and 2 h, with 0 h being at dusk. The treatments were as follows: 1) control, Ringer-Locke buffer; 2) leptin, 0.5 μg/kg BW; 3) ghrelin, 2.5 μg/kg BW; 4) orexin B, 0.3 μg/kg BW; 5) leptin antagonist, 50 μg/kg BW, then ghrelin, 2.5 μg/kg BW; and 6) leptin antagonist, 50 μg/kg BW, then orexin B, 0.3 μg/kg BW. Blood samples (5 mL) were collected at 15-min intervals for 6 h. The administration of leptin increased (P < 0.05) plasma concentrations of melatonin during short-day (ShD) photoperiods and decreased (P < 0.05) them during long-day (LD) photoperiods, whereas ghrelin decreased (P < 0.05) melatonin concentrations during ShD photoperiod, and orexin had no effect (P > 0.1). Leptin attenuated (P < 0.05) ghrelin concentrations relative to the concentration in controls during ShD. The plasma concentrations of orexin were reduced (P < 0.05) after leptin infusions during LD and ShD photoperiods; however, ghrelin had the opposite effect (P < 0.05) on orexin concentration. Orexin increased (P < 0.05) ghrelin concentrations during LD. Ghrelin and orexin concentrations were increased (P < 0.05) after leptin antagonist infusions. Our data provide evidence that the secretion of leptin, ghrelin, and orexin are seasonally dependent, with relationships that are subject to photoperiodic regulation, and that leptin is an important factor that regulates ghrelin and orexin releases in sheep.     

Adiponectin and leptin are related to fat mass in horses

Plasma concentrations of adiponectin and leptin were measured in 23 mature Standardbred mares (age: 10 ± 3 years) and 12 weanling fillies (10 Quarter Horse/Belgian crossbreds and two Quarter Horses; aged: 4 ± 3 months) to test the hypothesis that adipocytokines are proportional to adiposity in horses. Rump fat thickness was measured using B-mode ultrasound and percent body fat (% fat) calculated using a published formula for the estimation of fatness in horses. Plasma adiponectin and leptin were determined using radioimmunoassay. In the absence of purified equine adiponectin or leptin, results were expressed as human equivalents (HE) of immunoreactive (ir) adipocytokines.Plasma ir-leptin HE concentration was positively correlated (r = 0.543; P < 0.001) with percent body fat and (r = 0.556; P < 0.001) to fat mass in all horses. The plasma ir-leptin HE concentration was lower (P = 0.03) in weanlings (1.90 ± 0.34 ng/mL HE) than in mature mares (3.47 ± 0.50 ng/mL HE). The ratio of ir-adiponectin HE to ir-leptin HE was negatively correlated (r = −0.621; P < 0.001) to percent fat and (r = −631; P < 0.001) to fat mass in all horses. The ratio of ir-adiponectin HE to ir-leptin HE was higher (P < 0.001) in weanlings (3.04 ± 0.51) than mature mares (1.03 ± 0.13). These data suggest that leptin is proportional while adiponectin is inversely proportional to adiposity in horses.     

Cortisol treatment reduces ghrelin signaling and food intake in tilapia, Oreochromis mossambicus

It is well known that after a stressor, levels of plasma cortisol rise, inducing physiological changes within the animal that are directed toward maintaining homeostasis. Less well understood is the role of cortisol in regulating food intake in teleosts. This study investigated the effect of cortisol on food intake and regulation of the neuroendocrine appetite-stimulating hormones, neuropeptide Y (NPY) and ghrelin, in tilapia (Oreochromis mossambicus). Male and female tilapia were randomly assigned to one of the following treatments: unhandled control, vehicle-injected control, or cortisol (2 μg/g BW). Food intake was determined 24 h after injection during a 1-h feeding trial. Cortisol reduced food intake (P < 0.001). An identical study was conducted to measure the effects of 24-h cortisol treatment on the endocrine regulators of food intake. Cortisol reduced stomach expression of ghrelin mRNA (P < 0.05) and plasma concentrations of ghrelin (P < 0.05). In the hypothalamus/optic tectum cortisol reduced levels of GHSR1a-LR (biologically active ghrelin receptor) mRNA. In the telencephalon/preoptic area cortisol significantly reduced levels of NPY and GHSR1b-LR (biologically inactive ghrelin receptor) mRNA. These findings suggest that anorexigenic actions of cortisol may be mediated via two separate pathways: (1) reducing circulating ghrelin levels as well as GHSR1a-LR expression in the hypothalamus/optic tectum and/or (2) suppressing NPY expression in the telencephalon/preoptic area.     

Variation in plasma adrenocorticotropic hormone concentration and dexamethasone suppression test results with season, age, and sex in healthy ponies and horses

The purpose of this study was to evaluate the variation in plasma adrenocorticotropic hormone (ACTH) concentration and dexamethasone suppression test (DST) results with season, age, and sex in healthy, pony mares (n=15) and pony stallions (n=14) living under semiferal conditions and horse mares (n=10) living at pasture. Plasma ACTH concentrations were measured in September 2002, and in January, May, and September 2003. DSTs were performed in January and September 2003. Plasma ACTH concentrations in September 2002 and September 2003 were similar and were significantly greater than in January and May (P < .001). Plasma ACTH concentration was within the reference range for 38 (97%) of 39 subjects in January, for 39 (100%) of 39 subjects in May, for 2 (5%) of 39 subjects in September 2002, and for 3 (8%) of 39 subjects in September 2003. DST results were within the reference range in all subjects in January and were within the reference range for 29 (74%) of 39 subjects in September 2003. Plasma cortisol concentration at the end of the DST was significantly greater in September than in January (P = .002). Age was positively correlated with plasma ACTH and plasma cortisol concentration at the beginning and end of the DST Within the same season, plasma ACTH concentration in pony mares, pony stallions, and horse mares was not significantly different (P > .05). Seasonal changes in plasma ACTH concentration and DST results should be considered when interpreting endocrine test results.     

Effect of chronic clenbuterol administration and exercise training on immune function in horses

Effects of longitudinal exercise training and acute intensive exercise (simulated race test) on immune function have not been reported in horses. Clenbuterol, a beta2-adrenergic agonist, is used to manage inflammatory airway disease in horses. This study investigated the interaction of 8 wk of exercise training with or without 12 wk of clenbuterol administration in horses. Twenty-three untrained standardbred mares (10 +/- 3 yr, Mean +/- SE) were used and divided into four experimental groups. Horses given clenbuterol plus exercise (CLENEX; n = 6) and clenbuterol alone (CLEN; n = 6) received 2.4 microg/kg BW of clenbuterol twice daily (in an average volume of 20 mL) on a schedule of 5 d on and 2 d off for 12 wk. The CLENEX group was also aerobically trained 3 d/wk. Mares given exercise alone (EX; n = 5) were aerobically trained for 3 d/wk, and the control group (CON; n = 6) remained sedentary. Both EX and CON horses were administered similar volumes (approximately 20 mL) of molasses twice daily. A simulated race test (SRT) resulted in an elevation in lymphocyte number postexercise (P < 0.05). There was no significant difference after acute exercise in either monocyte or granulocyte number. Acute exercise resulted in a decrease (P < 0.05) in the percentage of CD4+ and an increase (P < 0.05) in the percentage of CD8+ cells. The SRT resulted in a decreased lymphoproliferative response to pokeweed mitogen (P < 0.05). A SRT had no effect on antibody production in response to equine influenza vaccine. The EX group demonstrated greater cortisol concentrations at rest and at all other time points postexercise after completing the training regimen compared with CLENEX horses (P < 0.05). Preexercise (SRT) peripheral blood monocyte number was lower in CLENEX horses than in other treatment groups (P < 0.05). Clenbuterol and exercise training did not significantly affect post-SRT changes in leukocyte numbers. Exercise training resulted in a decrease (P < 0.05) in the percentage of CD8+ cells post-SRT compared with other groups, but the percentage of CD4+ cells was not altered by either clenbuterol or exercise conditioning. Lymphocyte proliferative response was not affected by clenbuterol or exercise treatment. Horses demonstrated responses to bouts of acute exercise as noted with other species, namely humans and rodents.