Increase In Plasma Leptin Research Articles

β3-adrenoceptor mediates β3-selective agonist-induced effects on energy expenditure, insulin secrtion and food intake

Recent studies suggest that type 2 diabetes, which is usually associated with obesity, also involves defective energy expenditure, specifically fat oxidation in skeletal muscle. Skeletal muscle is a major site of fatty acid and glucose disposal. β-adrenoceptor (β-AR) agonists have previouslybeen shown to stimulate glucose uptake in isolated skeletal muscle, stimulate lipolysis and increase energy expenditure. This work was designed to study the β3-AR as a potential target of anti-diabetic control in rodents using BRL37344. Energy expenditure induced by BRL37344 inβ3-AR knockout mice and wild type litter mates as control, were studied. Mice were dosed with saline or BRL37344 (1mg/kg, i.p). Energy expenditure in mice was measured by open-circuit, indirect calorimetry over twenty four hours. In another set of mice, similar dose of BRL37344 orsaline (vehicle) were given. After three hours, mice were sacrificed and plasma collected for the determination of glucose, triglycerides, non-esterified fatty acids, insulin and leptin. BRL37344 did not induce a significant change in energy expenditure in both the β3-AR knockout mice and the wild type. Also the lipolytic effect of BRL37344 was non significant. However, there was a significant 52% decrease in plasma glucose in the knockout mice and a 42% decrease in the wild type. This was consistent with the four fold increase in insulin secretion in the knockout and atwo fold increase in the wild type. There was also a significant increase in plasma leptin in the knockout mice as compared to the wild type. Improved insulin sensitivity and decreased blood glucose, induced by BRL37344 imply β3-AR is a potential target for diabetes and obesity control, at least in the rodents.

Temporal and Site‐Specific Brain Alterations in CB1 Receptor Binding in High Fat Diet‐Induced Obesity in C57Bl/6 Mice

The cannabinoid CB1 receptor has been implicated in the regulation of appetite and the consumption of palatable foods. This experiment aimed to explore the involvement of the CB1 receptor in the early and late stages of high fat diet-induced obesity in C57BL/6 mice. The C57Bl/6 mice were placed on a high fat (HF) or low fat/high carbohydrate (LF) diet for 3 or 20 weeks. Quantitative autoradiography revealed that binding of [3H] CP-55,940 (CB1 receptor ligand) was elevated following 3 weeks of HF feeding in areas including the medial/ventral anterior olfactory nucleus (22.1%), agranular insular cortex (24.0%) and the hypothalamus (31.5%) compared to LF controls. This increased level of binding was correlated with an increase in plasma leptin in the hypothalamus, raising the possibility that this hormone may exert inhibitory control over endocannabinoid signalling at this stage of obesity. Mice fed a HF diet for 20 weeks were obese, hyperphagic and had decreased CB1 receptor binding levels in the substantia nigra (12.8%) and ventral tegmental area (17.1%) compared to LF controls. The low [3H] CP-55,940 binding density seen in these reward-related areas in the late stage of obesity may be indicative of increased endocannabinoid release due to the chronic HF diet consumption.

Positive energy balance is associated with accelerated muscle atrophy and increased erythrocyte glutathione turnover during 5 wk of bed rest

Background: Physical inactivity is often associated with positive energy balance and fat gain.Objective: We aimed to assess whether energy intake in excess of requirement activates systemic inflammation and antioxidant defenses and accelerates muscle atrophy induced by inactivity.Design: Nineteen healthy male volunteers were studied before and at the end of 5 wk of bed rest. Subjects were allowed to spontaneously adapt to decreased energy requirement (study A, n = 10) or were provided with an activity-matched diet (study B, n = 9). Groups with higher (HEB) or lower (LEB) energy balance were identified according to median values of inactivity-induced changes in fat mass (ΔFM, assessed by bioelectrical impedance analysis).Results: In pooled subjects (n = 19; median ΔFM: 1.4 kg), bed rest–mediated decreases in fat-free mass (bioelectrical impedance analysis) and vastus lateralis thickness (ultrasound imaging) were significantly greater (P < 0.03) in HEBAB (−3.8 ± 0.4 kg and −0.32 ± 0.04 cm, respectively) than in LEBAB (−2.3 ± 0.5 kg and −0.09 ± 0.04 cm, respectively) subjects. In study A (median ΔFM: 1.8 kg), bed rest–mediated increases in plasma leptin, C-reactive protein, and myeloperoxidase were greater (P < 0.04) in HEBA than in LEBA subjects. Bed rest–mediated changes of glutathione synthesis rate in eythrocytes (l-[3,3-2H2]cysteine incorporation) were greater (P = 0.03) in HEBA (from 70 ± 19 to 164 ± 29%/d) than in LEBA (from 103 ± 23 to 84 ± 27%/d) subjects.Conclusions: Positive energy balance during inactivity is associated with greater muscle atrophy and with activation of systemic inflammation and of antioxidant defenses. Optimizing caloric intake may be a useful strategy for mitigating muscle loss during period of chronic inactivity.

A cholecystokinin‐1 receptor agonist (CCK‐8) mediates increased permeability of brain barriers to leptin

Leptin regulates energy expenditure and body weight by acting both on the hypothalamus and on peripheral targets. Central actions of leptin are enhanced by cholecystokinin (CCK). The interaction between leptin and CCK makes physiological sense, as rats lacking CCK1 receptors are resistant to peripheral leptin but not to leptin directly infused into the brain. We have recently reported that CCK enhances leptin effects by increasing the entry of leptin into the CNS. The aim of this work was to further characterize the effect of CCK (10 microg kg(-1)) on leptin kinetics as well as the CCK receptor subtype involved in the interaction between CCK and leptin. Experiments were carried out both in free-feeding and in fasted rats receiving a single dose of leptin (100 microg kg(-1); i.p.). Parameters analysed over the next 6 h were plasma and cerebrospinal fluid concentrations of leptin. We observed that CCK-8 depressed the increase in plasma leptin that followed the i.p. injection and simultaneously increased leptin concentration in the cerebrospinal fluid from 92+/-25 to 230+/-24 pg mL(-1) (P<0.05). The effect of CCK-8 was totally prevented by the CCK1 receptor antagonist, SR-27,897 (0.3 mg kg(-1), s.c.), but not by the CCK2 receptor antagonist, L-365,260 (1 mg kg(-1)). These results show that CCK plays a role in regulating the access of leptin to the brain and suggest that CCK analogues, acting on CCK1 receptors, might be useful drugs in improving leptin actions within the brain.

Innate Immunity Modulates Adipokines in Humans

Chronic inflammation converges in type 2 diabetes and atherosclerosis. Modulation of adipokine signaling by innate immunity in humans is of considerable interest given the role of adipokines in insulin resistance and atherosclerosis. The aim of the study was to examine effects of low-grade endotoxemia, a model of human inflammation, on adipokines in vivo. An open-label, placebo-controlled, fixed-sequence clinical study was conducted at a General Clinical Research Center. There were 20 healthy male (50%) and female volunteers aged 18-40 yr. Serial blood sampling and adipose biopsies were performed for 24 h before and after iv bolus endotoxin [lipopolysaccharide (LPS), 3 ng/kg]. We measured plasma leptin, adiponectin, resistin, soluble leptin receptor, cytokines, insulin, and glucose; distribution of adiponectin among multimeric complexes; whole blood, monocyte and adipose mRNA for adipokines and their receptors. LPS induced fever, blood, and adipose TNF and IL-6 and increased homeostasis model assessment of insulin resistance. These were associated with increases in plasma leptin (from 4.1 +/- 1.1 to 6.1 +/- 1.9 ng/ml in men; 21.1 +/- 4.4 to 27.4 +/- 4.7 ng/ml in women; P < 0.005), doubling of the leptin:soluble leptin receptor ratio, and marked induction of whole blood resistin mRNA (13.7 +/- 7.3-fold; P < 0.001) and plasma resistin (8.5 +/- 2.75 to 43.2 +/- 15.3 ng/ml; P < 0.001). Although total adiponectin levels and low and high molecular weight adiponectin complexes were unaltered by LPS treatment, whole blood mRNA for adiponectin receptors 1 (49%; P < 0.005) and 2 (65%; P < 0.001) was suppressed. Modulation of adipokine signaling may contribute to the insulin resistant, atherogenic state associated with human inflammatory syndromes. Targeting of individual adipokines or their upstream regulation may prove effective in preventing acute and chronic inflammation-related metabolic complications.

48-h Glucose infusion in humans: Effect on hormonal responses, hunger and food intake

Experimentally-induced hyperglycemia by prolonged glucose infusion allows investigation of the effects of sustained stimulation of the pancreatic β-cell on insulin secretion and sensitivity. Hormonal responses to a meal following prolonged glucose infusions have not been investigated. To determine if a 48-h glucose infusion alters hormonal responses to a test meal as well as food intake and hunger in normal weight individuals, 16 subjects (8 men, 8 women, age 18–30 years, mean BMI = 21.7 ± 1.6 kg/m 2) were infused for 48 h with either saline (50 ml/h) or 15% glucose (200 mg/m 2/min). Subjects ingested a 600 kcal mixed nutrient meal 3 h after infusion termination. Blood samples were taken during the 48 h and for 4 h following food ingestion. The 48-h glucose infusion elicited a metabolic profile of a glucose intolerant obese subjects, with increased plasma glucose, insulin and leptin (all P < 0.01) and increased HOMA-IR ( P < 0.001). During meal ingestion, early insulin secretion was increased ( P < 0.05) but post-prandial glucose ( P < 0.01) and insulin ( P < 0.01) excursions were lower following the glucose infusion. Post-prandial plasma triglyceride concentrations were increased after glucose compared with saline. Food intake and hunger ratings were not different between the two conditions. Plasma leptin levels were inversely correlated with hunger ( P < 0.03) in both conditions and with food intake ( P < 0.003) during the glucose condition only. Thus, a 48-h glucose infusion does not impair post-prandial hormonal responses, alter food intake or hunger in normal weight subjects. The glucose-induced increases in plasma leptin result in a stronger inverse relationship between plasma leptin and hunger as well as food intake. These data are the first to demonstrate a relationship between leptin and hunger in normal weight, non-calorically restricted human subjects.

Developmental and Nutritional Regulation of the Prepubertal Bovine Mammary Gland: II. Epithelial Cell Proliferation, Parenchymal Accretion Rate, and Allometric Growth

It is well documented that elevated nutrient intake prior to puberty reduces prepubertal mammary development in the bovine. The companion paper demonstrated that age at harvest is a primary determinant of parenchymal (PAR) mass and that any effects of elevated energy intake on mechanisms regulating mammary development are dwarfed by this effect of time. Therefore, it is hypothesized that while causing a decrease in prepubertal PAR mass, elevated nutrient intake will have no effect on growth characteristics of the mammary gland. The objectives of this experiment were to evaluate the effects of increased nutrient intake from early in life on 1) mammary epithelial cell proliferation, 2) mammary PAR DNA accretion rates, and 3) the dynamics of prepubertal allometric PAR growth. Holstein heifers (n = 78) were fed from 45kg of body weight either elevated (E) or restricted (R) levels of nutrients to support 950 (E) or 650 (R) g/d of body weight gain. Six heifers per treatment were harvested at 50-kg increments from 100 to 350kg of body weight. Heifers on the E plane of nutrition had higher plasma leptin and less PAR DNA than their body weight-matched R-intake cohorts. Despite this reduction in PAR DNA, treatment did not negatively influence mammary epithelial cell proliferation or the PAR DNA accretion rate. Dynamics of allometric and isometric mammary growth were also unaffected by the level of nutrient intake, as was exit from allometric growth. This work represents the first demonstrating that the level of nutrient intake and the concomitant increase in plasma leptin have no measurable influence on 1) the rate of PAR DNA accretion, 2) mammary epithelial cell proliferation, or 3) total PAR mass and, by default, the local or systemic controls that coordinate these processes.

Gender-specific orexigenic and anorexigenic mechanisms in rats

Feeding dysregulation may manifest as either under-nourishment (e.g., anorexia) or excessive eating leading to obesity. Recent studies have suggested a gender-related variance in weight maintenance in response to chronic disease or obesity-related dietary regimens. However it is unclear whether these gender differences in weight management are secondary to appetite-mediated food intake or alternative mechanisms (e.g., exercise, metabolism). In this study, we explored gender-dependent feeding and hormonal responses to dietary restriction (12-h fast) or to an inflammatory stimulus (LPS, 100 μg/kg b.w.; i.p.) in rats. In response to a 12 h fast, female rats increased ( p < 0.05) total daily food intake above that of male rats by primarily increasing nighttime feeding by 40%, as compared to 10% in males. Consistent with the increased food intake, fasting induced a greater percent increase in female as compared to male plasma ghrelin (141 vs. 65%, p < 0.001). In response to LPS, both male and female rats showed similar reductions in total daily food consumption. However LPS (6 h) induced a greater percent increase in plasma leptin in female than male rats (230 vs. 33%, p < 0.01), whereas ghrelin was similarly decreased in both females and males (66 vs. 44%). These findings demonstrate sexual dimorphic responses in feeding and appetite-associated hormonal responses to fasting or LPS treatment. Our findings suggest that therapeutic interventions with ghrelin or leptin must be modified according to gender in order to optimally achieve either weight loss for obesity or weight gain/maintenance for chronic illness-associated anorexia.

Plasma leptin and ghrelin in the neonatal rat: interaction of dexamethasone and hypoxia

Ghrelin, leptin, and endogenous glucocorticoids play a role in appetite regulation, energy balance, and growth. The present study assessed the effects of dexamethasone (DEX) on these hormones, and on ACTH and pituitary proopiomelanocortin (POMC) and corticotropin-releasing hormone receptor-1 (CRHR1) mRNA expression, during a common metabolic stress - neonatal hypoxia. Newborn rats were raised in room air (21% O2) or under normobaric hypoxia (12% O2) from birth to postnatal day (PD) 7. DEX was administered on PD3 (0.5 mg/kg), PD4 (0.25 mg/kg), PD5 (0.125 mg/kg), and PD6 (0.05 mg/kg). Pups were studied on PD7 (24 h after the last dose of DEX). DEX significantly increased plasma leptin and ghrelin in normoxic pups, but only increased ghrelin in hypoxic pups. Hypoxia alone resulted in a small increase in plasma leptin. Plasma corticosterone and pituitary POMC mRNA expression were decreased 24 h following the last dose of DEX, whereas plasma ACTH and pituitary CRHR1 mRNA expression had already increased (normoxia and hypoxia). Hypoxia alone increased corticosterone, but had no effect on ACTH or pituitary POMC and CRHR1 mRNA expression. Neonatal DEX treatment, hypoxia, and the combination of both affect hormones involved in energy homeostasis. Pituitary function in the neonate was quickly restored following DEX-induced suppression of the hypothalamic-pituitary-adrenal axis. The changes in ghrelin, leptin, and corticosterone may be beneficial to the hypoxic neonate through the maintenance of appetite and shifts in intermediary metabolism.

Dysregulation of energy homeostasis in mice overexpressing insulin-like growth factor-binding protein 6 in the brain

IGFs, IGF receptors and IGF binding proteins (IGFBPs) are widely expressed in the central nervous system. To investigate the physiological significance of IGFBP-6 in the brain we established two transgenic mouse lines overexpressing human (h)-IGFBP-6 under the control of glial fibrillary acidic protein promoter. Increasing evidence suggests that insulin/IGF signalling pathways could be implicated in the neuroendocrine regulation of energy homeostasis. We explored the impact of brain IGFBP-6 overexpression on the regulation of food intake and energy balance. Transgenic mice were fed either a control diet or a high-fat diet for up to 3 months. Glucose and insulin tolerance tests were carried out before and after the diet period. Plasma parameters (insulin, leptin, glucose, NEFAs and triglycerides) were measured, and uncoupling protein 1 (UCP-1) expression was quantified in brown adipose tissue. Oxygen consumption was also measured in both groups. The transgenic mice fed a high-fat diet for 3 months developed obesity, showing increases in plasma leptin, glucose and insulin levels and mild insulin resistance. As compared with wild-type mice, no significant differences were found in the quantity of food intake. However, UCP-1 expression was down-regulated in the brown adipose tissue of the transgenic mice. Our results show that brain IGFBP-6 has an impact on the regulation of energy homeostasis. These transgenic h-IGFBP-6 mice may be considered a new tool for studies of the involvement of the brain IGF system in metabolism control and obesity.

Feeding-unrelated factors influencing the plasma leptin level in ruminants

The triglyceride content of lipid depots associated with the current feeding level is the primary determinant of leptin gene expression and the circulating leptin level. In laboratory rodents and primates the plasma leptin is influenced also by the age, gender and physiological status (puberty, pregnancy, lactation, postpartum period), and by the health condition such as sepsis due to Gram-negative (GN) bacteria. Some pathologic conditions with intensive cytokine release evoke an increase in plasma leptin, which is thought to depress the subsequent feed intake. However, the effect of these secondary factors may be species-dependent, with still unknown clinical relevance in ruminants. In our ovine and bovine models plasma leptin increased after castration and dexamethasone treatment, decreased after experimental administration of synthetic androgens in castrated rams, but remained unchanged throughout the ovarian cycle and after ovariectomy. The circulating leptin level increased temporarily during synthetic progestin (fluorogestone) treatment in ewes, but similar changes were not seen in progesterone-supplemented ewes and norgestomet-treated cows. In a second trial on dairy cows we wanted to study whether elevated plasma leptin levels are induced by experimental endotoxin mastitis, or by natural outbreak of GN mastitis and puerperal metritis. Experimental endotoxin mastitis resulted in some-hour elevation in cortisol and insulin, with a simultaneous decrease in IGF-I and thyroid hormones. In the first 14 days of lactation GN mastitis induced the same endocrine alterations as the experimental endotoxin challenge, but in natural cases these changes varied within a wider range, and were more protracted and robust. Cows with puerperal metritis had more obvious catabolic changes in the early weeks of lactation, than their healthy counterparts. However, both mastitis and puerperal metritis failed to increase the circulating leptin level, showing that in cows the plasma leptin is not responsible for the anorexia associated with these inflammatory diseases.

Effects of chronic obesity and weight loss on plasma ghrelin and leptin concentrations in dogs

The objective of this study was to evaluate, in dogs, the effects of obesity and weight loss on plasma total ghrelin and leptin concentrations. Twenty-four Beagle dogs, 12 control lean and 12 obese dogs of both genders and aged between 1 and 9 years, were used for the experiments. Mean body weight was 12.7 ± 0.7 kg for the lean group and 21.9 ± 0.8 kg for the obese group. The trial was divided into three phases. During phase 1, all 24 Beagle dogs were fed a maintenance diet. During phase 2, the obese dogs were submitted to a weight loss protocol with a high protein-low energy diet. The weight loss protocol ended once dogs reached optimal body weight. During phase 3, the dogs that were submitted to the weight loss protocol were maintained at their optimal body weight for 6 months. Plasma total ghrelin, leptin, insulin and glucose concentrations were measured to evaluate the effects of obesity and weight loss on these parameters in dogs. Body weight, body condition score, thoracic and pelvic perimeters, and ingested food amounts were also recorded during the study. Obese dogs demonstrated a significant decrease in plasma ghrelin and a significant increase in plasma leptin and insulin concentrations when compared with control dogs. During weight loss, significant increases in plasma total ghrelin and glucose and significant decreases in plasma leptin and insulin were observed. The increase in plasma ghrelin concentrations seemed to be transient. Body weight and the morphometric parameters correlated positively with leptin concentrations and negatively with total ghrelin concentrations. These results suggest that ghrelin and leptin could play a role in dogs in the adaptation to a positive or negative energy balance, as observed in humans.

Leptin, Fetal Nutrition, and Long-Term Outcomes for Adult Hypertension

One factor contributing to later hypertension, particularly in response to nutritional challenges is excess fat deposition around the kidney. In this review we discuss the hypothesis that these adverse conditions can be entrained by exposure of the conceptus to maternal nutrient restriction in early pregnancy. To this end we have shown in sheep that maternal nutrient restriction coincident with the time of embryogenesis and placental growth results in an early increase in fetal fat mass around the kidney that persists into later life. This is accompanied by an increase in leptin mRNA abundance and growth factor sensitivity. These adaptations occur in conjunction with reduced maternal plasma cortisol, thyroid hormones and leptin concentrations over the period of nutrient restriction. Some, but not all of these effects on fat development are accompanied by long term cardiovascular adaptations. As young adults, offspring from mothers nutrient restricted between early to mid gestation exhibit a leftward resetting, and blunting, of the cardiovascular baroreflex that appears to be mediated centrally through altered regional angiotensinogen II activity. At the same time, fat mass remains raised in nutrient restricted offspring. These animals demonstrate a marked increase in plasma leptin following sympathetic stimulation which is not observed in controls that indicates resetting of adipocyte sensitivity to stress. In conclusion, global nutrient restriction confined to the periods of embryonic and placental development therefore, programmes adult physiology, which may enhance predisposition to later disease given the appropriate environmental stimuli.

Pregnancy increases plasma leptin in nulliparous but not primiparous goats while lactation depresses it

Most dairy ruminants are still lactating during early pregnancy, which could induce hormonal adaptations different from those observed during pregnancy alone. The incidence of concomitant lactation and pregnancy on plasma leptin has not been studied, and physiological factors involved in its regulation have not been addressed in goats. We assayed leptinemia throughout the pregnancy–lactation cycle in nulliparous and primiparous goats, starting 165 days prior to parturition and finishing 59 days after. During the first half of pregnancy, primiparous goats were lactating. Lactating non-pregnant primiparous goats were studied in parallel. Plasma leptin increased (+49%) up to mid-pregnancy in nulliparous, but not in primiparous goats. Furthermore, leptinemia was similar between pregnant and non-pregnant lactating primiparous goats, suggesting a strong leptinemia down-regulation by late lactation. Plasma leptin decreased from mid-pregnancy to parturition, more markedly in nulliparous goats, and remained depressed during early lactation at a similar level in both female types. It was lower, at 130 days of pregnancy, in goats carrying two fetuses. The leptinemia down-regulation by late pregnancy was highlighted by the lack of plasma leptin increase after drying-off late-pregnant primiparous goats, while it strongly increased in non-pregnant goats. The observation of leptinemia increase only in nulliparous goats suggests that it is not an essential endocrine adaptation during early pregnancy. By contrast, in both female types, the low leptinemia during transition from late pregnancy to lactation, and during late lactation, may be important for the adaptations that occur during lactation such as the partitioning of energy and nutrients towards essential functions and/or hyperphagia.

Treatment with interleukin-11 affects plasma leptin levels in inflamed and non-inflamed rabbits

Treatment with the anti-inflammatory cytokine, interleukin-11 (IL-11), in rabbits with TNBS-colitis reduces tissue damage but does not normalize body weight loss despite an increase in plasma levels of motilin, known to stimulate food intake. We investigated whether IL-11 could increase plasma levels of the anorectic peptide, leptin in non-inflamed and inflamed rabbits. In addition, the effect of IL-11 and leptin on motilin mRNA expression in the T84 cell line was tested. Five days post-inflammation, weight loss amounted 10.7±1.2%, but plasma leptin and motilin levels were unaffected. During IL-11 treatment, weight loss remained and plasma leptin levels dose-dependently increased with 27±5% (4 μg/kg day) and 108±7% (720 μg/kg day). Motilin levels increased in parallel with 23±12% or 256±97%. In non-inflamed animals, a prompt decrease in weight (−11.9±1%) was observed after treatment with the highest dose of IL-11 and this was associated with an increase in plasma leptin (70±18%) and motilin levels (113±7%). Both IL-11 and leptin stimulated motilin mRNA expression in T84 cells with a different time profile. In conclusion, the increase in plasma leptin levels during IL-11 treatment induces wasting in normal rabbits and may be one of the major factors involved in the maintenance of body weight loss in rabbits with colitis. Increase of motilin expression by leptin may be part of a feedback mechanism.

PC47 IMPACT OF MATERNAL UNDERNUTRITION DURING EARLY-MID GESTATION ON RESULTING ADULT OFFSPRING IN SHEEP

Introduction: Suboptimal maternal nutrition during pregnancy is associated with increased risk of diseases in resulting adult offspring. Maternal undernutrition during placental growth produced a longer fetus at term with a disproportionately larger placenta and increased fat deposition. This study aimed to determine the long term consequences of early-mid gestation maternal undernutrition on blood pressure and plasma leptin concentration in young adult offspring. Methods: Seventeen male singleton bearing Scottish Black-face ewes of similar liveweight and body condition were individually housed from day of mating. Nine ewes were nutrient restricted (NR) consuming 4.0 MJ of metabolisable energy (ME) per day (50% ME requirements for maintenance and growth of conceptus) until 95 days gestation with 8 controls (C) consuming 8.0 MJ/day. After 95 days gestation until term (147 days) all ewes consumed 100% of requirements. At term lambs were delivered spontaneously and at 3 years old, the jugular vein and carotid artery were surgically catheterised for blood pressure (BP) measurement and blood sampling. Plasma was collected before and after a noradrenaline (NA) infusion with dose increases (2–48 ng/kg/min) at 10-minute intervals, and leptin concentration analysed. Animals were then humanely euthanased by barbiturate overdose to allow tissue sampling. Results are presented as mean ± standard error. Results: Between groups offspring body weight was similar at birth and 3 years of age but the NR group had larger fat (omental, perirenal and subcutaneous) deposits compared to controls. Basal BP prior to feeding was significantly higher in the NR offspring compared to controls (eg. diastolic 65 ± 3; NR 74 ± 2 mmHg (P<0.05)). Following NA infusion BP was similar between groups and rose with increasing NA concentration. With controls heart rate simultaneously decreased but this response was blunted in the NR group. The NR but not C offspring exhibited a significant increase in plasma leptin following NA infusion (C pre NA 3.22 ± 1.9; Post NA 3.16 ± 1.7; NR pre NA 3.47 ± 0.9; Post NA 4.5 ± 1.3 ng/ml (P<0.05)). Conclusion: Nutrient restriction between early-mid gestation produced hypertensive offspring with a blunted heart rate response to a vasoconstrictor challenge, with more fat and higher plasma leptin levels. Altering maternal nutrition during gestation may contribute to reprogramming of adult blood pressure and fat deposition contributing to hypertension and obesity in later life.

Leptin receptor expression increases in placenta, but not hypothalamus, during gestation in Mus musculus and Myotis lucifugus

In addition to effects on appetite and metabolism, the hormone leptin is required for reproduction in mammals. Maternal plasma leptin is increased above non-pregnant levels in all mammals thus far examined, including humans. The increase in plasma leptin appears to result in part from upregulation of adipose leptin secretion (e.g., in mice), or from production and secretion of leptin from the placenta (e.g., in humans and some bats). The placenta may also modulate maternal leptin levels via production of a plasma leptin-binding protein (mice, humans). Thus, the placenta plays a coordinating role in regulation of maternal leptin during pregnancy. In this study, the hypothesis that the placenta is also a target organ for leptin in diverse taxa was tested by examining the expression of leptin receptors (Ob-R) in placentae from species of distantly related mammalian taxa, Mus musculus (the laboratory mouse) and Myotis lucifugus (the little brown myotis, also called the little brown bat). A partial sequence of M. lucifugus Ob-R cDNA was first obtained and found to share ∼78–88% homology at the nucleotide level with known mammalian Ob-R cDNAs. Using probes and primers designed from this sequence, receptor expression was detected in numerous tissues of M. lucifugus, including placenta, which expressed two major receptor isoforms as judged by molecular size. In both species, Ob-R mRNA expression in placenta significantly increased from early to late gestation. Expression of Ob-R mRNA was not affected by cAMP treatment in vitro. The increase in Ob-R mRNA expression in placenta was specific, since Ob-R mRNA expression did not change during gestation in either species in hypothalamus, the major site of the central actions of leptin. Thus, Ob-R is expressed in placenta throughout gestation in mice and bats, and its expression increases over the course of gestation, which raises the possibility that leptin may exert temporally distinct effects on placental growth or function throughout gestation. Because similar placenta-specific changes in leptin receptor expression occurred in species from distantly related mammalian taxa which collectively comprise ∼70% of all known mammalian species, it is possible that placental actions of leptin are conserved across mammals, even in those species (such as the Swiss-Webster strain of mouse) in which the placenta does not itself produce leptin.

Role of endothelin-1 in blood pressure regulation in a rat model of visceral obesity and hypertension.

Endothelial dysfunction has been suggested to play an important role in the development of obesity-induced hypertension. Because endothelin release increases in response to endothelial damage, we examined whether endothelin-1 contributes to increased arterial pressure in a model of visceral obesity produced by feeding Sprague-Dawley rats a high-fat (HF) diet (40% fat w/w, n=6) for 12 months. Arterial and venous catheters were implanted for measurement of mean arterial pressure (MAP) and heart rate (HR) 24 hours per day and intravenous infusions. After a 5-day control period, rats were infused with the selective endothelin-1 type A receptor (ET-A) blocker ABT-627 (2.5 mg/kg per day, IV) for 9 days, followed by a recovery period. Rats fed a standard chow (normal fat, or NF, group: n=6) for 12 months were also infused with ET-A blocker and were used as controls. Compared with NF rats, HF rats had higher MAP (113+/-4 versus 98+/-2 mm Hg), increased visceral fat (18.7+/-2.0 versus 10.8+/-1.4 g), and 3.2-fold increase in plasma leptin despite similar total body weight gain. Long-term ET-A blockade markedly reduced MAP in HF (-14+/-3 mm Hg) and NF (-14+/-2 mm Hg), but it had no effect on HR, GFR, or PRA. These results indicate that a long-term HF diet may cause visceral obesity and increased MAP, even in the absence of major changes in total body weight. Endothelin-1 appears to play an important role in the maintenance of arterial pressure in rats fed HF and NF diets, but it does not appear to contribute to increased MAP in this model of diet-induced hypertension.

Effects of agouti-related protein on metabolism and hypothalamic neuropeptide gene expression.

The hypothalamic melanocortin system regulates feeding in part through interaction of the appetite stimulating peptide, agouti-related protein (AGRP), and the anorectic peptide, alpha-melanocyte stimulating hormone, a peptide derived from the pro-opiomelanocortin (POMC) polyprotein. Central administration of AGRP induces hyperphagia and increased gain in body weight in rodents, but may also exert metabolic effects even when hyperphagia is prevented. In the present studies, the effects of AGRP on hypothalamic neuropeptide gene expression and metabolism were examined in the rat. Central administration of AGRP for 3- and 7-day periods resulted in hyperphagia, increased body weight and increased plasma leptin and insulin concentrations compared to saline-injected controls. Hypothalamic concentrations of Pomc mRNA were also increased by 27% and 44% (in 3- and 7-day experiments, respectively). The hypothalamic concentration of Agrp mRNA was unchanged after 3 days, but was significantly decreased by 33% after 7 days of AGRP infusion. To determine if these changes were dependent upon AGRP-induced hyperphagia, pair-fed rats with restricted food intake receiving central administration of AGRP were also studied. In the absence of hyperphagia, intracerebralventricular administration of AGRP caused significant increases in plasma leptin and insulin concentrations (two-fold and 1.5-fold, respectively) and fat pad mass. A significant increase in hypothalamic Pomc mRNA concentrations was not detected in pair-fed rats. In contrast, Agrp mRNA concentrations remained suppressed by 45% in the pair-fed group after 7 days of AGRP infusion despite equal body weight compared to saline controls. The ratio of hypothalamic Pomc to Agrp mRNA was elevated two-fold in ad libitum and pair-fed AGRP-injected rats, which is consistent with increased stimulation of central melanocortin signalling pathways. Thus, central administration of AGRP exerts changes in hypothalamic neuropeptide gene expression and metabolic effects that are independent of the effects on food intake and body weight.

Nutritional and Developmental Regulation of Plasma Leptin in Dairy Cattle

Leptin is thought to play a critical role in regulating energy metabolism throughout mammalian life. In growing dairy cattle, plasma leptin has been proposed as a partial mediator of the effects of nutrition on reproductive and mammary development. However, the developmental stage at which the plane of nutrition increases plasma leptin has not been well defined. Further, it is unknown whether the onset of puberty is affected by plasma leptin concentration in dairy cattle. To investigate these questions, two studies were performed. In the first study, neonatal calves were fed a milk replacer at levels supporting an average daily gain of 570 g/d (L) or 1210 g/d (H). Weekly blood samples were obtained until slaughter at 105kg of body weight. Plasma leptin and adiposity remained constant in the L calves, but started to increase by the third week of age in the H calves. In the second study, 3- to 5-mo-old heifers were fed a total mixed ration supplemented with either calcium salts of palm fat or conjugated linoleic acids at levels sustaining an average daily gain of approximately 1.0 kg/d. Blood samples were obtained until the third postpubertal luteal phase. The fat source had no effects on growth parameters, body composition, age at puberty, or plasma leptin. Therefore, plasma leptin was reanalyzed as a function of age from start of treatment until slaughter. The plasma concentration of leptin remained nearly constant at 2.3ng/ml until 1 yr of age, when a rise in plasma leptin became obvious. Puberty occurred with equal frequency either around 1 yr of age when plasma leptin was nearly constant or later when leptin was rising rapidly. We conclude that plasma leptin is regulated by nutrition in early postnatal life, but that a sudden increase in plasma leptin is not required for the onset of puberty in dairy cattle.