Reproduction and beyond,kisspeptin in ruminants

Kisspeptin(Kp) is synthesized in the arcuate nucleus and preoptic area of the hypothalamus and is a regulator of gonadotropin releasing hormone in the hypothalamus.In addition,Kp may regulate additional functions such as increased neuropeptide Y gene expression and reduced proopiomelanocortin(POMC) gene expression in sheep.Other studies have found a role for Kp to release growth hormone(GH),prolactin and luteinizing hormone(LH)from cattle,rat and monkey pituitary cells.Intravenous injection of Kp stimulated release LH,GH,prolactin and follicle stimulating hormone in some experiments in cattle and sheep,but other studies have failed to find an effect of peripheral injection of Kp on GH release.Recent studies indicate that Kp can stimulate GH release after intracerebroventricular injection in sheep at doses that do not release GH after intravenous injection.These studies suggest that Kp may have a role in regulation of both reproduction and metabolism in sheep.Since GH plays a role in luteal development,it is tempting to speculate that the ability of Kp to release GH and LH is related to normal control of reproduction.     

Reproduction and beyond,kisspeptin in ruminants

Kisspeptin (Kp) is synthesized in the arcuate nucleus and preoptic area of the hypothalamus and is a regulator of gonadotropin releasing hormone in the hypothalamus. In addition, Kp may regulate additional functions such as increased neuropeptide Y gene expression and reduced proopiomelanocortin (POMC) gene expression in sheep. Other studies have found a role for Kp to release growth hormone (GH), prolactin and luteinizing hormone (LH) from cattle, rat and monkey pituitary cells. Intravenous injection of Kp stimulated release LH, GH, prolactin and follicle stimulating hormone in some experiments in cattle and sheep, but other studies have failed to find an effect of peripheral injection of Kp on GH release. Recent studies indicate that Kp can stimulate GH release after intracerebroventricular injection in sheep at doses that do not release GH after intravenous injection. These studies suggest that Kp may have a role in regulation of both reproduction and metabolism in sheep. Since GH plays a role in luteal development, it is tempting to speculate that the ability of Kp to release GH and LH is related to normal control of reproduction.     

Growth hormone releasing factors and secretion of growth hormone in sheep,calves and pigs

Human pancreatic growth hormone releasing factors (hpGRF) (1–40) and (1–44) were administered iv in sheep, pigs and calves to determine their effectiveness in stimulating GH release in these species. Both peptides produced a rapid increase in plasma GH concentration in all three species at dose levels ranging from .0065 to .65 nmol/kg. Moreover, there was no difference in the GH-secretory response observed between hpGRF(1–44)NH₂ and (1–40)OH in sheep. Sheep also responded to hpGRF(1–40)NH₂ and (1–40)OH as well as [his₁]- and [tyr₁]-hpGRF(1–40)NH₂ in a similar manner. Rat hypothalamic GRF was less effective than [his₁]-hpGRF(1–40), while the response to bGRF was not significantly different from hpGRF(1–40) in stimulating GH secretion in sheep. Although all three species responded to hpGRF, the elevation in plasma GH levels above baseline were greater after hpGRF injection in sheep than in pigs or calves. Subcutaneous injection of hpGRF in sheep was an effective mode of administration of the peptide, although the effect was not as long-lasting as that after iv injections and higher doses were required to stimulate GH secretion.     

The neurophysiological regulation of growth hormone secretion

With the advent of genetic engineering, the importance of GH in the regulation of growth and metabolism in domestic species has been clearly demonstrated. Ample evidence of an integral role for GH in the processes of growth and lactation exists in dairy cattle (1,2), sheep (3), beef cattle (4) and swine (5). For example, circulating GH levels are high during the period of rapid growth in several species including cattle (6), swine (7) and poultry (8). Endogenous GH secretion is primarily controlled by the central nervous system (CNS) via two specific hypothalamic neurohormones, growth hormone-releasing factor (GRF) and somatostatin (SRIF), an inhibitor of GH release. The secretion of GRF and SRIF is governed by a host of neuropeptides and neurotransmitters which provide a functional link between higher CNS centers and hypophysiotropic neurons. This review will focus on the CNS regulation of GH secretion and circulating factors which feedback to either stimulate or inhibit its release.     

Neuroendocrine regulation of growth hormone secretion in sheep. III. Serotoninergic systems.

The role of serotoninergic pathways in the regulation of growth hormone secretion in the sheep has been investigated. Both peripheral and central routes of administration of serotonin agonists and antagonists have been used. Intravenous administration of the serotonin agonist, buspirone, at 1.2 mg/kg/h lowered plasma GH levels (P less than 0.001) but at 0.21 mg/kg/h there was no significant decrease. Intracerebroventricular (icv) administration of serotonin itself also depressed GH levels (P less than 0.01). The serotonin antagonist, cyproheptadine, failed to affect GH concentrations when given either intravenously (0.25 mg/kg/h) or intracerebroventricularly (4 mg). Neither serotonin nor cyproheptadine had any significant effect on plasma glucose or cortisol levels when administered icv. The possible role of somatostatin in mediating the serotonin associated decrease in GH was investigated by concurrent administration of serotonin and a specific, potent anti-somatostatin serum into a cerebral ventricle. This treatment also resulted in a marked, sustained depression in GH (P less than 0.001). These data suggest that serotonin can inhibit release of GH from the pituitary in sheep and that this is independent of hypothalamic somatostatin.     

Ghrelin differentially modulates the GH secretory response to GHRH between the fed and fasted states in sheep

The effect of energy balance on the growth hormone (GH) secretory responsiveness to growth hormone-releasing hormone (GHRH) has not been determined in ruminant animals. Therefore, we examined the effects of intravenous injections of 0, 3.3, and 6.6 μg ghrelin/kg body weight (BW), with and without GHRH at 0.25 μg/kg BW, on GH secretory responsiveness in both the fed and fasted sheep. The injections were carried out at 48 h (Fasting state) and 3 h (Satiety state) after feeding. Blood samples were taken every 10 minutes, from 30 minutes before to 120 minutes after the injection. Low (3.3 μg/kg BW) and high (6.6 μg/kg BW) doses of ghrelin stimulated GH secretion significantly (P < .05) greater in the Satiety state than in the Fasting state. Growth hormone-releasing hormone plus both doses of ghrelin stimulated GH secretion significantly (P < .05) greater in the Satiety state than in the Fasting state. Ghrelin and GHRH exerted a synergistic effect in the Satiety state, but not in the Fasting state. Plasma ghrelin levels were maintained significantly (P < .05) greater in the Fasting state than in the Satiety state except the temporal increases after ghrelin administration. Plasma free fatty acid (FFA) concentrations were significantly (P < .01) greater in the Fasting state than in the Satiety state. In conclusion, the present study has demonstrated for the first time that ghrelin differentially modulates GH secretory response to GHRH according to feeding states in ruminant animals.     

α- and β-adrenergic influences on metabolism in heart and skeletal muscle

Using fenylephrine, isoprenaline, noradrenaline, adrenaline and receptor blockers, adrenergic regulation of metabolism in working heart as well as in skeletal resting muscle of rats was investigated. Changes were determined in: phosphocreatine level and³²P incorporation into phosphocreatine; phosphorolytic and hydrolytic activities of glycogenolysis; glycogen synthetase activity and¹⁴C-glucose incorporation into glycogen; and the level of cAMP. It was found that, contrary to the β-receptor, stimulation of the α-adrenergic receptor inhibits catabolism of macroenergetic phosphates and diminishes glycogen decomposition on the phosphorolytic pathway while increasing the hydrolysis of glycogen and glycogen synthetase activity as well as glucose incorporation into glycogen. On the basis of these data a hypothesis may be put forward that the influence of the α-adrenergic receptor results in the diminishing of energy utilization and in the activation of glycogen anabolism in heart and skeletal muscle. An adrenergic intrasystemic receptor antagonism in the regulation of energy consumption by heart and skeletal muscle is indicated. The α-receptor seems to be an adrenergic intrasystemic moderator of the metabolism rate which is accelerated by β-receptor influence. This suggests that besides sympathetico-parasympathetic intersystemic antagonism regulating the heart work and thereby cardiac energy consumption, there is a separate adrenergic mechanism directly controlling the balance between catabolism and anabolism in heart muscle. This mechanism, based on α- and β-adrenergic receptor antagonism, also controls the metabolic balance in skeletal muscles.     

Regulation of the growth hormone and luteinizing hormone response to endotoxin in sheep

Infectious disease processes cause physiological adaptations in animals to reorder nutrient partitioning and other functions to support host survival. Endocrine, immune and nervous systems largely mediate this process. Using endotoxin injection as a model for catabolic disease processes (such as bacterial septicemia), we have focused our attention on regulation of growth hormone (GH) and luteinizing hormone (LH) secretion in sheep. Endotoxin produces an increase in plasma GH and a decrease in plasma LH concentrations. This pattern can be reproduced, in part, by administration of various cytokines. Antagonists to both interleukin-1 (IL-1) and tumor necrosis factor (TNF) given intravenously (IV) prevented the endotoxin-stimulated increase in GH. Since endotoxin will directly stimulate GH and LH release from cultured pituitary cells, the data suggest a pituitary site of action of the endotoxin to regulate GH. Studies with portal vein cannulated sheep indicated that gonadotropin releasing hormone was inhibited by endotoxin, suggesting a central site of action of endotoxin to regulate LH. However, other studies suggest that endotoxin may also regulate LH secretion at the pituitary. Thus, IL-1 and TNF regulate GH release from the pituitary gland while endotoxin induces a central inhibition of LH release.     

Neuroendocrine regulation of growth hormone secretion in sheep. II. Effect of somatostatin on growth hormone and glucose levels.

The effect of intravenous (iv) and intracerebroventricular (icv) administration of somatostatin on the plasma levels of growth hormone (GH) and glucose was studied in sheep. Intravenous somatostatin decreased (P less than 0.001) circulating GH when infused at the rate of 5 micrograms/min (150 ng/kg/min) over 1 hr, but when used at 1 microgram/min there was no effect on plasma GH levels during infusion. At both doses used there was an indication of an increase in GH following the cessation of somatostatin infusion. Somatostatin given at both these doses iv had no effect on plasma glucose levels. When given icv neither 1.8 micrograms, 18 micrograms nor 180 micrograms somatostatin had any significant effect of plasma GH levels, although there was a significant (P less than 0.05) elevation in GH levels 75 min after 180 micrograms somatostatin icv. Plasma glucose levels did not increase following injection of somatostatin icv at 1.8 or 18 micrograms, but there was a clear hyperglycaemic episode following 180 micrograms icv. Despite a lack of effect of somatostatin on GH release when given icv, there was a clear elevation (P less than 0.05) in plasma GH levels immediately following icv administration of a somatostatin antiserum. These data indicate that iv administration of somatostatin at pharmacological levels can depress unstimulated GH levels in sheep while administration icv does not. Central administration of somatostatin increases plasma glucose levels only at high doses and seems unlikely to be of physiological importance in glucose homeostasis.     

Growth hormone-releasing hormone and somatostatin concentrations in the hypophysial portal blood of conscious sheep during the infusion of growth hormone-releasing peptide-6

The effects of growth hormone-releasing peptide-6 (GHRP-6) on peripheral plasma concentrations of growth hormone (GH) and hypophysial portal plasma concentrations of growth hormone-releasing hormone (GHRH) and somatostatin (SRIF) were investigated in conscious ewes. Paired blood samples were collected from the hypophysial portal vessels and from the jugular vein of nine ewes for at least 2 hr. The sheep were then given a bolus injection of 10 μg of GHRP-6 per kg followed by a 2-hr infusion of GHRP-6 (0.1 μ/kg · hr). Blood sampling continued throughout the infusion and for 2 hr afterwards. An increase in plasma GH concentration was observed in the jugular samples of six of the nine ewes (1.4 ± 0.3 vs 7.4 ± 2.0 ng/ml, P < 0.05) 5–10 min after the GHRP-6 bolus injection, but in no case did we observe a significant coincident release of GHRH. During the infusion period, mean plasma GHRH levels were not significantly increased but there was a 50% increase (P < 0.05) in GHRH pulse frequency; GHRH pulse amplitude was not changed. Mean SRIF concentration, pulse frequency, and pulse amplitude were unchanged by GHRP-6 treatment. These data indicate that GHRP-6 causes a small, but significant effect on the pulsatile secretion of GHRH, indicating action at the hypothalamus or higher centers of the brain. The large initial GH secretory response to GHRP-6 injection does not appear to be the result of GHRP-6 action on GHRH or SRIF secretion.     

Neuroregulation of growth hormone secretion in domestic animals

Growth hormone (GH) is essential for postnatal somatic growth, maintenance of lean tissue at maturity in domestic animals and milk production in cows. This review focuses on neuroregulation of GH secretion in domestic animals. Two hormones principally regulate the secretion of GH: growth hormone-releasing hormone (GHRH) stimulates, while somatostatin (SS) inhibits the secretion of GH. A long-standing hypothesis proposes that alternate secretion of GHRH and SS regulate episodic secretion of GH. However, measurement of GHRH and SS in hypophysial-portal blood of unanesthetized sheep and swine shows that episodic secretion of GHRH and SS do not account for all episodes of GH secreted. Furthermore, the activity of GHRH and SS neurons decreases after steers have eaten a meal offered for a 2-h period each day (meal-feeding) and this corresponds with reduced secretion of GH. Together, these data suggest that other factors also regulate the secretion of GH. Several neurotransmitters have been implicated in this regard. Thyrotropin-releasing hormone, serotonin and gamma-aminobutyric acid stimulate the secretion of GH at somatotropes. Growth hormone releasing peptide-6 overcomes feeding-induced refractoriness of somatotropes to GHRH and stimulates the secretion of GHRH. Norepinephrine reduces the activity of SS neurons and stimulates the secretion of GHRH via alpha(2)-adrenergic receptors. N-methyl-D,L-aspartate and leptin stimulate the secretion of GHRH, while neuropeptide Y stimulates the secretion of GHRH and SS. Activation of muscarinic receptors decreases the secretion of SS. Dopamine stimulates the secretion of SS via D1 receptors and inhibits the secretion of GH from somatotropes via D2 receptors. Thus, many neuroendocrine factors regulate the secretion of GH in livestock via altering secretion of GHRH and/or SS, communicating between GHRH and SS neurons, or acting independently at somatotropes to coordinate the secretion of GH.     

Neuroendocrine regulation of growth hormone secretion in sheep. IV. Central and peripheral cholecystokinin

The result of alterations in the levels of CCK, in the blood and in the cerebrospinal fluid, on the functioning of the growth hormone axis has been examined in sheep. Male Coopworth sheep of about 40 kg liveweight were given various doses of CCK either intracerebroventricularly (icv) or intravenously (iv). Other similar sheep were given various doses of a CCK antagonist (loxiglumide) by the same routes. Bolus iv administration of either 35 μg or 200 μg of CCK had no effect on plasma GH levels. When given icv, however, CCK resulted in a marked (P<0.01) prolonged depression in plasma GH levels. The decrease in GH secretion could be partially attenuated by concurrent administration of loxiglumide, but was completely unaffected by concurrent administration of anti-somatostatin serum icv. Loxiglumide alone had no effect on plasma GH levels when given at up to 200 μg icv, but intravenous administration of 8 mg of the CCK antagonist resulted in an increase in plasma GH concentrations (P<0.05). Plasma levels of somatostatin, glucose and cortisol were unaffected by both icv and iv administration of CCK. These results show that CCK can have a strong GH-inhibiting effect in the brain. Furthermore, this effect seems to be independent of hypothalamic somatostatin, suggesting another GH-inhibiting system exists.     

Growth hormone secretion and pancreatic function following somatostatin infusion in ducks (Anas platyrhynchos)

The intravenous infusion of somatostatin (800 ng/kg min) reduced the concentration of growth hormone (GH) in the plasma of 4 to 5, 6 to 7 and 8 to 9 week-old ducklings, but not in adult ducks. The inhibition of GH secretion was not due to accompanying changes in pancreatic function, since the infusion of a lower dose of somatostatin (200 ng/kg min) increased glucagon release and decreased plasma free fatty acids (FFA), as observed with the higher dose, but had no effect on GH concentrations. The withdrawal of somatostatin inhibition resulted in rebound GH secretion in immature birds, the magnitude of which was directly related to the pre-treatment level. Following somatostatin infusion (800 ng/kg min) no modification in GH concentration was observed in adult ducks. These results demonstrate that basal GH release in young birds is not autonomous and is suppressible by somatostatin. The data provide further evidence for age-related changes in the control of avian GH and insulin release and for the independence of the effects of somatostatin on the pituitary and pancreas glands.     

The control of adenohypophysial hormone secretion by amino acids and peptides in swine

Several different amino acids and peptides control secretion of adenohypophysial hormones and this control may be indirect, via the modulation of hypothalamic hormone secretion. Indeed, classical hypothalamic hormones (e.g., gonadotropin-releasing hormone [GnRH], growth hormone-releasing hormone [GHRH], somatostatin, etc.) may be released into the hypothalamo-hypophysial portal vasculature, travel to the adenohypophysis and there stimulate or inhibit secretion of hormones. Alternatively, some amino acids and peptides exert direct stimulatory or inhibitory effects on the adenohypophysis, thereby impacting hormone secretion. In swine, the most extensively studied modulators of adenohypophysial hormone secretion are the excitatory amino acids (ExAA), namely glutamate and aspartate, and the endogenous opioid peptides (EOP). In general, excitatory amino acids stimulate release of luteinizing hormone (LH), follicle-stimulating hormone (FSH), growth hormone (GH), and prolactin (PRL). Secretion of adenohypophysial hormones induced by ExAA is primarily, but perhaps not exclusively, a consequence of action at the central nervous system. By acting primarily at the level of the central nervous system, EOP inhibit LH secretion, stimulate GH release and depending on the animal model studied, exert either stimulatory or inhibitory influences on PRL secretion. However, the EOP also inhibited LH release by direct action on the adenohypophysis. More recently, peptides such as neuropeptide-Y (NPY), orexin-B, ghrelin, galanin, and substance P have been evaluated for possible roles in controlling adenohypophysial hormone secretion in swine. For example, NPY, orexin-B, and ghrelin increased basal GH secretion and modulated the GH response to GHRH, at least in part, by direct action on the adenohypophysis. Secretion of LH was stimulated by orexin-B, galanin, and substance P from porcine pituitary cells in vitro. Because the ExAA and various peptides modulate secretion of adenohypophysial hormones, these compounds may play an important role in regulating swine growth and reproduction.     

Neuroendocrine regulation of growth hormone secretion in sheep. V. Growth hormone releasing factor and thyrotrophin releasing hormone.

The effects of intravenous (IV) and intracerebroventricular (ICV) administration of either bovine growth hormone releasing hormone (GRF) or thyrotrophin releasing hormone (TRH) on plasma growth hormone (GH) and glucose levels have been examined in sheep. Intravenous GRF 1-29NH2 at 3 and 30 micrograms stimulated an increase in GH levels in a dose-dependent fashion; administration of GRF into a lateral cerebral ventricle, however, produced a smaller GH response which was similar at these two doses. Evaluation of somatostatin levels in petrosal sinus blood (which collects pituitary effluent blood) showed that ICV administration of GRF stimulated a release of somatostatin into the blood. Furthermore, concurrent administration of GRF and a potent anti-somatostatin serum ICV resulted in a much enhanced release of GH which was similar to that obtained with a comparable dose of GRF given IV. TRH (as another putative GH-secretagogue) was also administered both IV and ICV. When given IV, 200 micrograms (but not 100 micrograms) TRH produced an elevation in GH levels. By contrast, when 5 micrograms TRH was given ICV there was a decrease in circulating GH levels, but no change in plasma somatostatin concentrations. These results indicate that the smaller GH response to ICV- compared with IV-administered GRF is due to the release of somatostatin within the brain. In addition, it would seem that TRH is not a physiological GH-secretagogue in sheep.     

Influence of age and sex on basal secretion of growth hormone (GH) and on GH-induced release by porcine GH-releasing factor pGRF(1–29NH2) in growing pigs

The aim of this study was to determine the effect of age and sex on basal secretory patterns of growth hormone (GH) and growth hormone-releasing factor (GRF) induced GH release. Eighteen pigs (9 castrated males and 9 females) were stimulated with pGRF(1–29)NH₂ at 7,11,15,19 and 23 weeks of age. Blood samples were taken from each animal via jugular vein cannulate every 20 min, from 6 hr before to 5 hr after iv GRF administration at a dose of 4 μg/kg. GH baseline levels, amplitude of the GH peaks, area under the GH peaks and the overall mean of GH serum levels decreased (P<.001) with age in both sexes. Age also had a marked effect on GRF-induced GH release: the amplitude of GH peaks and area under the GH peaks decreased (P<.001) with age. The GH response to pGRF(1–29)NH₂ varied considerably, depending on the timing of the episodic endogenous secretion of GH. An immediate response (<30 min) was observed when GRF was injected at the end of a trough period or at the beginning of a peak, but there was no immediate response when GRF was injected at the end of a peak or at the beginning of a trough period. Our results show that both endogenous GH secretion and pGRF(1–29)NH₂-induced GH release declines with age, suggesting a decreased sensitivity of the somatotroph cells to GRF with age; and that the high variability of the GH response to pGRF(1–29)NH₂ stimulation depends greatly on the timing of the episodic endogenous GH release, thus implying a possible episodic endogenous somatostatin secretion by the hypothalamus.     

Effects of ruminal infusion of volatile fatty acids on plasma concentration of growth hormone and insulin in sheep.

In an initial experiment we observed postprandial changes in plasma concentrations of growth hormone (GH), insulin, glucagon, and somatostatin (SRIF) in sheep. We then examined whether increasing the rumen concentration of volatile fatty acids (VFA) by infusing a VFA mixture at three rates (53.5, 107, and 214 micromol/kg/min for 4 hr) mimicked the postprandial changes in hormone secretion. Feeding significantly (P < 0.05) suppressed the plasma GH concentration for 6 hr, whereas it significantly (P < 0.05) increased plasma concentrations of insulin, glucagon, and SRIF. Plasma glucose levels tended to decrease after feeding but then gradually increased over the prefeeding level (P < 0.05). Intraruminal infusion of the VFA mixture at 107 micromol/kg/min caused similar changes in ruminal VFA concentrations to those seen after feeding. The infusion significantly (P < 0.05) suppressed GH secretion in a dose-dependent manner, whereas it caused a significant (P < 0.05) increase in insulin and glucose concentrations without changing glucagon concentrations. From these results, we conclude that the postprandial change in ruminal VFA concentration may be a physiological signal which modifies GH and insulin secretion in sheep.     

Recombinant Porcine Leptin Reduces Feed Intake and Stimulates Growth Hormone Secretion in Swine

Two experiments (EXP) were conducted to test the hypothesis that porcine leptin affects GH, insulin-like growth factor-I (IGF-I), insulin, thyroxine (T₄) secretion, and feed intake. In EXP I, prepuberal gilts received intracerebroventricular (ICV) leptin injections. Blood was collected every 15 min for 4 hr before and 3 hr after ICV injections of 0.9% saline (S; n = 3), 10 μg (n = 4), 50 μg (n = 4), or 100 μg (n = 4) of leptin in S. Pigs were fed each day at 0800 and 1700 hr over a 2-wk period before the EXP. On the day of the EXP, pigs were fed at 0800 hr and blood sampling started at 0900 h. After the last sample was collected, feeders were placed in all pens. Feed intake was monitored at 4, 20, and 44 hr after feed presentation. In EXP II, pituitary cells from prepuberal gilts were studied in primary culture to determine if leptin affects GH secretion at the level of the pituitary. On Day 4 of culture, 10⁵ cells/well were challenged with 10⁻¹², 10⁻¹⁰, 10⁻⁸, or 10⁻⁶ M [Ala¹⁵]-h growth hormone-releasing factor-(1-29)NH₂ (GRF), 10⁻¹⁴, 10⁻¹³, 10⁻¹², 10⁻¹¹, 10⁻¹⁰, 10⁻⁹, 10⁻⁸, 10⁻⁷, or 10⁻⁶ M leptin individually or in combinations with 10⁻⁸ and 10⁻⁶ M GRF. Secreted GH was measured at 4 hr after treatment. In EXP I, before injection, serum GH concentrations were similar. Serum GH concentrations increased (P < 0.01) after injection of 10 μg (21 ± 1 ng/ml), 50 μg (9 ± 1 ng/ml), and 100 μg (13 ± 1 ng/ml) of leptin compared with S (1 ± 2 ng/ml) treated pigs. The GH response to leptin was greater (P < 0.001) in 10 μg than 50 or 100 μg leptin-treated pigs. By 20 hr the 10, 50, and 100 μg doses of leptin reduced feed intake by 53% (P < 0.08), 76%, and 90% (P < 0.05), respectively, compared with S pigs. Serum IGF-I, insulin, T₄, glucose, and free fatty acids were unaffected by leptin treatment. In EXP II, relative to control (31 ± 2 ng/well), 10⁻¹⁰, 10⁻⁸, and 10⁻⁶ M GRF increased (P < 0.01) GH secretion by 131%, 156%, and 170%, respectively. Only 10⁻⁶ M and 10⁻⁷ M leptin increased (P < 0.01) GH secretion. Addition of 10⁻¹¹ and 10⁻⁹ M leptin in combination with 10⁻⁶ M GRF or 10⁻¹¹ M leptin in combination with 10⁻⁸ M GRF-suppressed (P < 0.05) GH secretion. These results indicate that leptin modulates GH secretion and, as shown in other species, leptin suppressed feed intake in the pig.     

Effect of atropine and pyridostigmine on growth hormone response to GH-releasing peptide-2 and GH-releasing hormone in swine

To investigate the effects of high and low somatostatinergic tone on GH-releasing peptide-2 (GHRP-2) and GH-releasing hormone (GHRH)-induced growth hormone (GH) secretion in swine, we examined GHRP-2- and GHRH-induced GH secretion after pretreatment with atropine or pyridostigmine. Pretreatment of swine with atropine (80 µg/kg bodyweight (BW), intravenous (i.v.)) 15 min before i.v. administration of saline, GHRP-2 (30 µg/kg BW), GHRH (1 µg/kg BW) or a combination of GHRP-2 and GHRH, reduced plasma GH area under the curve ( P  < 0.05), completely blocked GH response to GHRH, and attenuated GH response to GHRP-2 and GHRH combined ( P  < 0.05), without affecting GH response to GHRP-2 only. A synergistic effect of GHRP-2 and GHRH was not observed. In contrast, pretreatment of swine with pyridostigmine (100 µg/kg BW, i.v.), under the same pretreatment conditions as above, increased plasma GH concentration ( P  < 0.01), augmented GH response to GHRP-2 ( P  < 0.05), and GHRP-2 and GHRH combined ( P  < 0.05), but did not affect GH response to GHRH. These results suggest that the cholinergic muscarinic agents atropine and pyridostigmine modulate the GH response to GHRP-2 and GHRH, and that GHRP-2 acts antagonistically on the inhibitory effect of somatostatin in swine.     

Relationships between plasma concentrations of leptin and other metabolic hormones in GH-transgenic sheep infused with glucose

To study the regulation of leptin secretion in sheep, we infused glucose (0.32 g/h/kg for 12 h) into GH-transgenic animals (n = 8) that have chronically high plasma concentrations of ovine GH and insulin, but low body condition and low plasma leptin concentrations, and compared the responses with those in controls (n = 8). In both groups, the infusion increased plasma concentrations of glucose and insulin within 1 h and maintained high levels throughout the infusion period (P < 0.0001). Compared with controls, GH-transgenics had higher concentrations of insulin, IGF-1, GH (all P < 0.0001) and cortisol (P < 0.05), but lower GH pulse frequency (P < 0.0001). Overall, leptin concentrations were lower in GH-transgenics than in controls (P < 0.01). A postprandial increase in leptin concentrations was observed in both groups, independently of glucose treatment, after which the values remained elevated in animals infused with glucose, but returned to basal levels in those infused with saline, independently of transgene status. In both GH-transgenics and controls, glucose infusion did not affect the concentrations of GH, IGF-1, or cortisol. In conclusion, GH-transgenic and control sheep show similar responses to glucose infusion for leptin and other metabolic hormones, despite differences between them in body condition and basal levels of these hormones. Glucose, insulin, GH, IGF-1 and cortisol are probably not major factors in the acute control of leptin secretion in sheep, although sustained high concentrations of GH and IGF-1 might reduce adipose tissue mass or inhibit leptin gene expression.