Sex-specific epigenetic alterations of the hypothalamic Agrp-Pomc system do not explain 'diabesity' in the offspring of high-fat diet (HFD) overfed maternal rats.

Maternal high-fat diet (HFD) overfeeding pre- and during pregnancy and lactation may 'program' a 'diabesity' predisposition in the offspring, for inconclusive reasons. Acquired alterations of the hypothalamic promoter methylation and mRNA expression of the satiety neurohormone Pomc are possibly of critical importance here. We investigated within one developmental approach, including male and female rats, the sex-specific DNA methylation pattern and corresponding mRNA expression of both Pomc and its endogenous functional antagonist Agrp in the hypothalamus of adult HFD offspring. Obesity and diabetic disturbances occurred in both male and female HFD offspring, accompanied by altered Pomc promoter methylation pattern. However, this was not related to significant Pomc mRNA expression alterations. In contrast, male-specific alterations of Agrp promoter methylation were found, even associated with reduced mRNA expression of this orexigenic/anabolic Pomc antagonist. In conclusion, acquired epigenetic alterations of the hypothalamic Agrp-Pomc system hardly explain the 'diabesity' phenotype in HFD offspring, while distinct vulnerability and functionality of Agrp promoter and related genomic regions methylation should be further investigated.

Hypothalamic insulin receptor expression and DNA promoter methylation are sex-specifically altered in adult offspring of high-fat diet (HFD)-overfed mother rats.

Maternal overnutrition around reproduction has been shown to increase the offspring's risk for "diabesity," mediated by altered hypothalamic neuropeptide expression. In this report, a possible contribution of altered hypothalamic sensing capacity for the peripheral satiety signals glucose, insulin and leptin will be addressed, taking into account potential sex differences. Specifically, we evaluated the effects a maternal high-fat diet (HFD) overfeeding has in rats pre- and during pregnancy and lactation on the hypothalamic gene expression patterns of insulin and leptin receptors (InsR, ObRb) and glucose transporter 3 (Glut3) as well as DNA methylation in the offspring at adult age (day 200 of life). Maternal HFD consumption resulted in a metabolic syndrome phenotype, i.e., obesity, hyperleptinemia, hyperinsulinemia, impaired glucose tolerance and increased homeostatic model assessment of insulin resistance. Interestingly, in turn, insulin resistance was more pronounced in male offspring, accompanied by decreased hypothalamic InsR-mRNA. This was linked with hypermethylation of an activating transcription factor binding site within the hypothalamic InsR promoter. The degree of methylation correlated inversely with respective InsR expression, while InsR expression itself was inversely related to phenotypic "diabesity." Expression of ObRb and Glut3 mRNA was not significantly changed. In conclusion, sex-specific alterations of hypothalamic InsR expression and DNA promoter methylation in adult offspring of HFD-overfed dams may lead to hypothalamic insulin resistance and "diabesity," with males predisposed to this epigenetic malprogramming.

Temporary prenatal hyperglycemia leads to postnatal neuronal 'glucose-resistance' in the chicken hypothalamus.

Prenatal exposures may have a distinct impact for long-term health. Exposure to maternal 'diabesity' during pregnancy increases offspring 'diabesity' risk, e.g. by malprogramming the central nervous regulation of body weight, food intake and metabolism. Critical mechanisms and concrete disrupting factors still remain unclear. Due to the independent development, from the mother, the chicken embryo could provide a valuable model to distinctively establish causal factors. Aim of this study was to determine effects of temporary prenatal hyperglycemia on postnatal hypothalamic neuronal glucose sensitivity in the chicken. To induce hyperglycemia in chicken embryos, 0.5 ml glucose solution (concentration 30 mmol/l) were daily administered via catheter into a vessel of the chorioallantoic egg membrane from days 14 to 17 of incubation. On day 21 of postnatal age, body weight, body fat content, blood glucose, neuroelectrophysiological glucose sensitivity as well as glucose transporter expression were determined in hypothalamic brain slices. No significant changes in morphometric and metabolic parameters were observed. However, strongly decreased neuronal glucose sensitivity and glucose transporter expression occurred, indicating prenatally acquired hypothalamic 'glucose-resistance'. In conclusion, temporary late prenatal hyperglycemia induces lasting changes in central glucose sensing. The prenatally glucose-treated chicken provides a valuable new model for investigating early central nervous origins of 'diabesity' and related disorders.

Acquired alterations of hypothalamic gene expression of insulin and leptin receptors and glucose transporters in prenatally high-glucose exposed three-week old chickens do not coincide with aberrant promoter DNA methylation.

BACKGROUND: Prenatal exposures may have a distinct impact for long-term health, one example being exposure to maternal 'diabesity' during pregnancy increasing offspring 'diabesity' risk. Malprogramming of the central nervous regulation of body weight, food intake and metabolism has been identified as a critical mechanism. While concrete disrupting factors still remain unclear, growing focus on acquired epigenomic alterations have been proposed. Due to the independent development from the mother, the chicken embryo provides a valuable model to distinctively establish causal factors and mechanisms. AIM: The aim of this study was to determine the effects of prenatal hyperglycemia on postnatal hypothalamic gene expression and promoter DNA methylation in the chicken. METHODS AND FINDINGS: To temporarily induce high-glucose exposure in chicken embryos, 0.5 ml glucose solution (30 mmol/l) were administered daily via catheter into a vessel of the chorioallantoic egg membrane from days 14 to 17 of incubation. At three weeks of postnatal age, body weight, total body fat, blood glucose, mRNA expression (INSR, LEPR, GLUT1, GLUT3) as well as corresponding promoter DNA methylation were determined in mediobasal hypothalamic brain slices (Nucleus infundibuli hypothalami). Although no significant changes in morphometric and metabolic parameters were detected, strongly decreased mRNA expression occurred in all candidate genes. Surprisingly, however, no relevant alterations were observed in respective promoter methylation. CONCLUSION: Prenatal hyperglycemia induces strong changes in later hypothalamic expression of INSR, LEPR, GLUT1, and GLUT3 mRNA. While the chicken provides an interesting approach for developmental malprogramming, the classical expression regulation via promoter methylation was not observed here. This may be due to alternative/interacting brain mechanisms or the thus far under-explored bird epigenome.

Avoiding the prenatal programming effects of glucocorticoids: are there alternative treatments for the induction of antenatal lung maturation?

BACKGROUND: The long-term outcomes of antenatal glucocorticoids (GCs) vary between reports, and have generated controversy in terms of repeated and single-course events, causing irreversible effects on endocrine set points. AIM: This study aimed to assess the effects of alternative therapeutic agents other than synthetic glucocorticoid GC administration for fetal lung maturation. METHODS: A review of literature from PubMed, EMBASE, Cochrane Library, and Google Scholar was conducted to assess the use of alternative therapies to synthetic GCs using recommendations of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement (PRISMA). End points included the rates of respiratory distress syndrome (RDS), mRNA expression for pneumocyte type II, concentration of surfactant proteins in alveolar lavage, morphological differences, histological proof of lung maturation, and angiogenesis or quantification of the surfactant pool. RESULTS: In all 41 studies examined, we found that ambroxol showed positive effects on lung maturation, but it has yet to be analyzed with sufficient significance in humans. Interleukins and TNF-alpha produce accelerated lung maturation, but have only been evaluated in basic research/experimental studies. Growth factors promote structural and functional growth in all phases of lung maturation, but little is known about their reciprocal effects and exact mechanisms as therapeutics. Thyroid releasing hormone or vitamin A cause detrimental side effects or were less effective for lung maturation. CONCLUSIONS: The efficacy and safety of these alternative agents are differentiated and none up to now can be recommended as an alternative to GCs.

Detection of long-term influence of prenatal temperature stimulation on hypothalamic type-II iodothyronine deiodinase in juvenile female broiler chickens using a novel immunohistochemical amplification protocol.

It has been clearly shown that early environmental stimulation may have long-lasting influence on body functions. Because of the strong relationship between thermoregulation and other homeostatic linked physiological parameters, perinatal thermal manipulation will also have an impact on other body functions like reproduction. As a maturation stimulant for later reproductive performance, hypothalamic type-2 iodothyronine deiodinase (Dio2) expression was investigated in 35day old immature female broilers with and without embryonic temperature stimulation. For the first time, human-specific Dio2 primary antibodies combined with additional amplification enabled the immunohistochemical detection of hypothalamic Dio2 protein in birds. The novel protocol includes an additional amplification step involving swine-anti-rabbit/mouse/goat antibodies against both goat anti-Dio2 primary and rabbit anti-goat biotinylated secondary commercial antibodies in the standard diaminobenzidine protocol. However, significant Dio2 expression was exclusively found in perinatally short-term temperature stimulated hens. Caudal but not rostral hypothalamic slices revealed that elevating incubation temperature by 1°C for 2h daily, from day 18 of embryonic development until hatching, induced a statistical significant expression of Dio2 within the subcommisural organ and the median eminence. This ample expression of Dio2 protein within caudal but not rostral hypothalamic slices of embryonic temperature stimulated chickens, leads to the assumption of a novel physiological prospective for embryonic thermal manipulation involving the suppression of thyroid hormone and the boosting of hypothalamic Dio2-induced FSH secretion to considerably advance the age of photoinduced egg production. It could be also of practicable relevance for broiler breeder females, and needs further investigations.

Early postnatal life as a critical time window for determination of long-term metabolic health.

Epidemiological studies demonstrated a clear phenomenological association between low birth weight and increased cardiometabolic risk later in life, very similar to that in high birth weight subjects. Pre- and/or neonatal overfeeding appears to be an etiological clue. In animal studies, irrespective of birth weight neonatal over-nutrition leads to later overweight, impaired glucose tolerance and cardiometabolic alterations. Probably, perinatally acquired alterations of DNA methylation patterns of gene promoters of central nervous regulators of body weight and metabolism play a key role in mediating these relationships. In humans, the long-term impact of neonatal nutrition is conclusively demonstrated by studies on the consequences of breastfeeding vs. formula-feeding. Taken together, the quantity and quality of nutrition during neonatal life plays a critical role, beyond prenatal development, in the long-term programming of health and disease. This opens a variety of opportunities and challenges to primarily prevent chronic diseases, e.g. the metabolic syndrome.

Short-term regulation of the hypothalamic melanocortinergic system under fasting and defined glucose-refeeding conditions in rats: a laser capture microdissection (LMD)-based study.

It is well established that under fasting conditions the expression of the orexigenic neuropeptide agouti-related peptide (AGRP) is up-regulated in the hypothalamic arcuate nucleus (ARC), while inconsistent data exist regarding fasting regulation of the anorexigenic neurohormone proopiomelanocortin (POMC). Inconsistencies might have methodological reasons, especially concerning neuromorphological and/or experimental (nutritional) specificity. We analyzed the expression of both neuropeptides in ARC neurons, using lasercapture microdissection (LMD) and real-time PCR in 12h fasted vs. fed Wistar rats as well as after a standardized glucose load, i.e., under clinically relevant conditions in terms of diagnosing glucose intolerance in the human. Under fasting conditions, clear up-regulation of AGRP was observed, with increasing magnitude in ARC single neurons (SNP) as compared to ARC cell layers (+125% vs. +23%, resp.), closely correlated to hypoinsulinemia and hypoleptinemia. Surprisingly, in the fasting state POMC was not found to be down-regulated, neither in ARC cell layers nor in ARC single neurons (+9% vs. +6%). However, glucose-refeeding under diagnostically relevant conditions led to strong neuronal up-regulation of POMC expression in ARC SNP (+128%), and AGRP down-regulation (-50%). In conclusion, experimentally, topographically, and analytically specific and standardized conditions confirmed AGRP in ARC neurons as being neuronally up- and down-regulated, resp., depending on the general nutritional state, while POMC was found to be (up-) regulated only after peripheral glucose load. Findings suggest that POMC in ARC neurons acts glucose-mediated as an "anti-orexigenic" neurohormone, specifically responding to hyperglycemia.

Leptin determination in colostrum and early human milk from mothers of preterm and term infants.

BACKGROUND: Leptin is involved in the regulation of food intake and energy expenditure and is therefore important for growth and brain development. Analytical methods used for leptin measurement in human milk differ widely in the literature and yield varying results. AIMS: To compare different preparation methods for the analysis of leptin in human milk and to investigate the leptin levels in colostrum and mature human milk from mothers of preterm or term infants. METHODS: Mothers delivering a preterm (n=37) or a term infant (n=40) were recruited for a prospective study and were ask to collect breast milk on the 3rd and 28th day of lactation. Leptin, protein and fat concentrations were analysed. Clinical data of mother and child were recorded prospectively. RESULTS: Skim milk was most appropriate for leptin analysis. Human milk leptin concentrations did not differ between preterm and term human milk. In term milk, leptin concentration on day 28 was lower than on day 3 (p<0.05). Milk leptin levels on the 3rd and 28th day were positively correlated with mothers' body mass index, but not with fat content in milk. CONCLUSION: Skim milk was the most stabile preparation for leptin analysis. Preterm and term human milk contain leptin in equal concentrations. Human milk leptin depends on mothers' body mass index.

Epigenetic malprogramming of the insulin receptor promoter due to developmental overfeeding.

AIM: Prenatal and neonatal overfeeding programs a permanent obesity and diabetes disposition, e.g., due to induction of hypothalamic insulin resistance. We investigated acquired alterations of the DNA methylation pattern of the hypothalamic insulin receptor promoter (IRP) which might be an underlying molecular mechanism. METHODS: Neonatal overfeeding was induced by rearing Wistar rats in small litters (SL). Methylation of CpG-dinucleotides of the hypothalamic IRP was mapped using bisulfite sequencing. RESULTS: Neonatal overfeeding led to rapid early weight gain, resulting in a metabolic syndrome phenotype, i.e., obesity, hyperleptinemia, hyperglycemia, hyperinsulinemia, and increased insulin/glucose-ratio. The proportion of animals carrying any methylated CpG residue in the 322 bp CpG island of the IRP was increased in neonatally overfed SL rats (n=8), as compared to controls (n=8; P=0.04). Moreover, the mean percentage of methylated CpG positions was also higher in SL rats (P=0.01). Over both groups, neonatal blood glucose levels were positively correlated to the extent of promoter methylation (r=0.52; P=0.04). CONCLUSIONS: This study characterizes for the first time the IRP epigenomically in any species and tissue. Our data reveal that the IRP is vulnerable to hypermethylation due to overnutrition, probably especially glucose-dependent in a dose-response manner. This paradigmatically indicates the impact of nutrient-dependent epigenetic malprogramming, leading to a "diabesity" disposition which may become pathogenic throughout life.

Hypothalamic proopiomelanocortin promoter methylation becomes altered by early overfeeding: an epigenetic model of obesity and the metabolic syndrome.

Pre- and neonatal overfeeding programmes a permanent obesity disposition and accompanying diabetic and cardiovascular disorders, by unknown mechanisms. We proposed that early overfeeding may alter DNA methylation patterns of hypothalamic promoter regions of genes critically involved in the lifelong regulation of food intake and body weight. We induced neonatal overfeeding by rearing Wistar rats in small litters (SL) and thereafter mapped the DNA methylation status of CpG dinucleotides of gene promoters from hypothalamic tissue, using bisulfite sequencing. Neonatal overfeeding led to rapid early weight gain, resulting in a metabolic syndrome phenotype, i.e. obesity, hyperleptinaemia, hyperglycaemia, hyperinsulinaemia, and an increased insulin/glucose ratio. Accompanying, without group difference to controls, the promoter of the main orexigenic neurohormone, neuropeptide Y, was methylated at low levels (i.e. < 5%). In contrast, in SL rats the hypothalamic gene promoter of the main anorexigenic neurohormone, proopiomelanocortin (POMC), showed hypermethylation (P < 0.05) of CpG dinucleotides within the two Sp1-related binding sequences (Sp1, NF-kappaB) which are essential for the mediation of leptin and insulin effects on POMC expression. Consequently, POMC expression lacked upregulation, despite hyperleptinaemia and hyperinsulinaemia. Accordingly, the extent of DNA methylation within Sp1-related binding sequences was inversely correlated to the quotients of POMC expression/leptin (P = 0.02) and POMC expression/insulin (P < 0.001), indicating functionality of acquired epigenomic alterations. These data for the first time demonstrate a nutritionally acquired alteration of the methylation pattern and, consequently, the regulatory 'set point' of a gene promoter that is critical for body weight regulation. Our findings reveal overfeeding as an epigenetic risk factor of obesity programming and consecutive diabetic and cardiovascular disorders and diseases, in terms of the metabolic syndrome.

Action of prolactin, prolactin-releasing peptide and orexins on hypothalamic neurons of adult, early postnatally overfed rats.

OBJECTIVES: Hypothalamic neurons of rats overweight due to early postnatal overfeeding (SL) differ from those of control rats in their responses to feeding relevant hormones like leptin or insulin. The question arose whether prolactin and prolactin-releasing peptide (PrRP) express also differential action in SL rats. These peptides are described to have an effect on food intake and body weight regulation. Prolactin is co-synthesized in lateral hypothalamic neurons together with orexins that were also analyzed in this study. METHODS: Single unit activity was extracellularly recorded in brain slices from adult control rats (CL) and from rats previously raised in small litters (SL). The action of the peptides on the firing rates was evaluated in the medial parvicellular part of the paraventricular nucleus (PaMP) and the medial arcuate nucleus (ArcM). RESULTS: In control rats, PrRP significantly activated PaMP neurons, whereas prolactin and orexin-A induced also inhibition. In SL rats, there was a significantly different effect of orexin-B on PaMP neurons: the main effect changed from activation in controls to inhibition. ArcM neurons of both control and SL rats were mainly excited by prolactin and orexins. CONCLUSION: Changes acquired during early development of neuronal responses to feeding relevant peptides are not a general non-specific mechanism of neurochemical plasticity, but concern specific hypothalamic nuclei and/ or hormones and neuropeptides. The increase in inhibition by orexin-B of hypothalamic paraventricular neurons could in vivo contribute to the neonatally acquired disposition towards persistingly increased food intake and reduced energy expenditure of overweight SL rats.

Perinatal programming and functional teratogenesis: impact on body weight regulation and obesity.

It is increasingly accepted that alterations of the intrauterine and early postnatal nutritional, metabolic, and hormonal environment may cause predispositions for the development of diseases in later life. Studies in the offspring of diabetic mothers have decisively contributed to this perception. Alterations of the fetal and neonatal environment which offspring of diabetic mothers 'experience' seem to program a disposition to develop obesity, diabetes mellitus and Syndrome X-like alterations throughout later life. Underweight at birth is also suggested to lead to an increased risk of Syndrome X in later life ('Barker hypothesis'). Pathophysiological mechanisms are unclear. Hormones are important environment-dependent organizers of the developing neuro-endocrine-immune network, which finally regulates all fundamental processes of life. When present in non-physiological concentrations during 'critical periods' of perinatal life, induced by alterations in the intrauterine or neonatal environment, hormones can act as 'endogenous functional teratogens'. Perinatal hyperinsulinism is pathognomonic in the offspring of diabetic mothers. Early hyperinsulinism also occurs as a result of early postnatal overfeeding. In rats, endogenous hyperinsulinism, as well as peripheral or only intrahypothalamic insulin treatment during perinatal development, may lead to 'malprogramming' of neuroendocrine systems regulating body weight, food intake and metabolism. This results in an increased disposition to become obese and to develop diabetes throughout life. In conclusion, a complex malprogramming of the central regulation of body weight and metabolism may provide a general etiopathogenetic concept, explaining perinatally acquired dispositions, thereby opening a wide field of primary prevention.

The main effect of cocaine- and amphetamine-regulated transcript (CART) peptide on hypothalamic neuronal activity depends on the nutritional state of rats.

OBJECTIVES: The anorectic and catabolic action of CART is primarily mediated by the hypothalamus. The study proved the hypothesis that neurons of the hypothalamic regulatory system of body weight differentially react to CART in dependence of the nutritional state of the animal: overweight, fed or short-term fasting. DESIGN AND SETTING: Single unit activity was extracellularly recorded in brain slices. The action of CART was studied in brains of 1. overweight adult rats previously subjected to early postnatal overfeeding in small litters (SL), compared to control litters, 2. normal rats that were deprived of food for 24 h, compared to fed rats. RESULTS: Hypothalamic dorsomedial neurons of controls, but not SL rats were significantly excited by CART, ventromedial neurons of SL rats were significantly inhibited. Also neurons of hungry rats were significantly inhibited. MAIN FINDINGS: Controls and overweight SL as well as fed and hungry rats differed significantly in the neuronal effects of CART. The predominant effect of the peptide did neither depend on weight nor on age of animals, but on neonatal development or nutritional state. CONCLUSION: The increase in inhibition by CART of ventromedial and dorsomedial neuronal activity could in vivo contribute to increased food intake and reduced energy expenditure of overweight SL as of hungry rats. Since leptin is able to change synaptic wiring and the expression of excitatory and inhibitory synapses already within short time, the increased expression of inhibitory responses to CART may reflect a general mechanism in adaptation of neuronal regulatory systems to the nutritional state, in fed, adult small-litter rats acquired during the postnatal critical differentiation period, thus leading to permanently altered function.

Hypothalamic neurons of postnatally overfed, overweight rats respond differentially to corticotropin-releasing hormones.

Adult overweight rats previously subjected to early postnatal overnutrition in small litters are hyperphagic, hyperleptinemic and differ in emotional behaviour from rats of control litters. We proved the hypothesis that neurons of the hypothalamic regulatory system of body weight differentially react to peptides of the corticotropin-releasing factor (CRF) family in these overweight rats. Single unit activity was recorded in brain slices. In controls, CRF and the CRF(2) receptor agonist stresscopin-related peptide (SRP) predominantly activated neurons of the dorsomedial part of ventromedial hypothalamic nucleus (VMHDM), but in overweight rats, SRP induced a significant inhibition. Increased neuronal firing to CRF and SRP of the medial parvocellular part of paraventricular hypothalamic nucleus (PaMP) in controls similarly changed to more inhibition in overweight rats. Inhibition of neuronal activity in VMHDM and PaMP can contribute to reduce satiety signals and to decrease energy expenditure in rats. In contrast, medial arcuate (ArcM) neurons of controls were significantly inhibited by SRP, whereas neurons of overweight rats could also be activated. The difference in the expression of these response types was significant. Activation of ArcM neurons known to produce neuropeptide Y can increase food intake. The results are discussed in terms of a trophic action of leptin changing synaptic wiring and the expression of excitatory and inhibitory synapses. The altered responses of hypothalamic neurons in adult small-litter rats may reflect a general mechanism of neurochemical plasticity acquired during the postnatal critical differentiation period, thus leading to permanently altered function of the regulatory system of body weight.

Altered action of dopamine and cholecystokinin on lateral hypothalamic neurons in rats raised under different feeding conditions.

Single-unit activity was recorded in the lateral hypothalamus (LH) of adult Wistar rats anaesthetized with urethane. The rats were differently nourished till weaning by raising in small (SL), control (CL) or large litters (LL). They gained significantly different body weight leading to overweight in SL (mean: 428.4 g on day 90) and underweight in LL rats (mean 399.5 g) compared to CLs (414.5 g). The mean basal firing rate of LH neurons differed, it was lowest in SL and highest in LL rats. The proportion of neurons changing their firing rate by more than 30% in response to iontophoretically administered dopamine (DA) was significantly greater in SL (76%) than LL rats (54%). Effects of DA were significantly more often blocked by a D1 receptor antagonist in LL than CLs. The responsiveness to cholecystokinin (CCK) alone and coadministered with DA was also greater in SL than LL. Furthermore, the proportion of neurons inhibited by DA alone and in the presence of CCK was significantly greater in SL than LL rats. In conclusion, litter size and difference in nourishment during early postnatal development of rats seem to determine LH basal firing rate. The increased neuronal responsiveness to exogenous DA and CCK in neonatally overfed SL rats may indicate a decreased activity of these endogenous signals which normally contribute to limitation of energy intake.

Altered neuronal responses to feeding-relevant peptides as sign of developmental plasticity in the hypothalamic regulatory system of body weight.

Neuronal plasticity during the critical postnatal period of development seems to promote a change in the function of the hypothalamic regulatory system of body weight. Rats raised in small litters (SL) of only three pups per mother compared to ten or twelve in control litters (CL) gain significantly more weight than normal rats till weaning and are overweight also in later life. These rats are known to express hyperleptinemia, hyperglycemia and hyperinsulinemia. The review summarizes the results of action of leptin and insulin as well as of several feeding-relevant neuropeptides on neuronal activity of hypothalamic regulatory centres in overweight SL rats compared to controls. The study was performed on brain slices perfused with solution containing 10 mM glucose. Whereas a normally inhibitory action of leptin and insulin on medial arcuate neurons (ArcM) is reduced in SL rats and partly replaced by activation, the normally activating effect of these hormones on ventromedial (VMH) neurons is altered to predominant inhibition. Inhibition of ArcM neurons may decrease the release of the orexigenic neuropeptide Y (NPY) and agouti gene-related protein (AGRP). Thus, the negative feedback by leptin and insulin on food intake is replaced by diminished response and partly positive feedback processes in SL rats. The action of NPY and AGRP as well as of the orexigenic melanin-concentrating hormone on paraventricular (PVH) and VMH neurons is also shaped from activation or bimodal effects to predominant inhibition. Such inhibition of PVH and VMH might lead to reduced energy expenditure in small litter rats. Also the anorexigenic melanocortin alpha-MSH seems to contribute into increased energy storage. These altered responses of hypothalamic neurons in overweight small litter rats might reflect a general mechanism of neurochemical plasticity and "malprogramming" of hypothalamic neuropeptidergic systems leading to a permanently altered regulatory function.