Histamine H1-receptors differentially mediate the action of amylin on hypothalamic neurons in control and in overweight rats.

The hypothalamic arcuate, dorsomedial and paraventricular nuclei are involved in regulation of body weight and food intake and contain binding sites for the anorexigenic amylin. Effects of amylin on medial arcuate and paraventricular neurons studied in adult rats overweight through early postnatal overfeeding in small litters (SL) differed from those of control litters (CL). Now we observed that also dorsomedial neurons respond differentially to this satiety signal. They were significantly inhibited by amylin in SL but not CL rats. Since the histaminergic system seems to be involved in mediating effects of amylin, we studied the role of histamine H(1)-receptors. Single unit activity was recorded in brain slices of CL and SL rats in each of the three hypothalamic nuclei. The histamine H(1)-receptor antagonist pyrilamine differentially altered or reduced responses to amylin, not depending on the kind of litter but on the functional effect of the peptide. Pyrilamine prevented significant inhibition of medial arcuate neurons in controls as well as inhibition of dorsomedial and paraventricular neurons in SL rats. Searching for further mechanisms possibly contributing to the change of neuronal responses we found that in the presence of a GABA(A)-receptor antagonist amylin induced a significant inhibition of medial arcuate neurons in SL rats similar to that in CL without antagonist. Activation of medial arcuate neurons expressing the orexigenic neuropeptide Y and inhibition of dorsomedial and paraventricular neurons in SL rats may in vivo contribute to hyperphagia and overweight. Histamine H(1)-receptors and GABA(A)-receptors seem to be differentially involved in mediation of these effects.

Insulin resistance of hypothalamic arcuate neurons in neonatally overfed rats.

Rats exposed to early postnatal overfeeding by rearing in small litters become hyperphagic, hyperleptinemic, and hyperinsulinemic throughout later life. Medial arcuate neurons are involved in body weight regulation. They were tested in brain slices of control and small-litter rats concerning differences in responses to insulin. Insulin induced suppression of firing in controls, whereas in small-litter rats inhibition was significantly reduced and activation increased. This could be observed in juvenile as well as adult rats. A gamma-aminobutyric acid type A receptor antagonist did not change the responses. Thus, negative feedback to the satiety signal insulin on medial arcuate neurons is reduced in neonatally overfed small-litter rats. This can be regarded as insulin resistance, which is induced during early development and persists in later life.

GABAA receptor antagonists prevent abnormalities in leptin, insulin and amylin actions on paraventricular hypothalamic neurons of overweight rats.

The hypothalamic regulatory system of body weight which develops in rats during critical periods of early postnatal life seems to express plastic changes depending on nutrition at that time. Adult rats previously exposed to early postnatal overnutrition by raising them in small litters become persistently predisposed to overweight, hyperphagia and hyperleptinaemia. The hypothesis was raised that feeding-related peptides could be involved through altered effects on neuronal activity of the regulatory systems of such rats. This was studied on brain slices of small-litter rats and normal-weight controls between days 60 and 120 of life. Neurons of the medial parvocellular part of the paraventricular nucleus were significantly activated by the adiposity signals leptin, insulin and amylin in controls. This is a kind of negative feedback, because activation of these neurons is known to be followed in vivo by increased energy expenditure. GABAergic mechanisms seem to affect these neuronal responses because the activating effects of insulin and amylin were reduced in the presence of a GABA(A) receptor antagonist. In overweight small-litter rats, however, the neuronal responses to the adiposity signals were significantly changed; activating effects were reduced and inhibitory effects increased. By means of blockade of GABA(A) receptors, significant alterations in the neuronal responses to leptin, insulin and amylin in small-litter rats were prevented. Responses to the peptides were reversed and now resembled those of controls. In conclusion, changes in neuronal wiring with GABAergic interneurons seem to contribute to a persistently reduced negative feedback of adiposity signals in early postnatally overfed rats.

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.

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.

Arcuate neurons of overweight rats differ in their responses to amylin from controls.

Amylin and ghrelin are hormones produced, respectively, in pancreas and stomach. They have a central action on food intake and body weight. Possible changes in their effect on hypothalamic neuronal activity were investigated in overweight rats previously subjected to early postnatal overnutrition compared to controls. Single unit activity of brain slices was recorded in the medial arcuate that contains neuropeptide Y producing neurons. The orexigenic ghrelin activated neurons of both groups of rats. The anorexigenic amylin, however, significantly inhibited neurons of controls but not of overweight rats. The difference in the type of response to amylin shown by arcuate neurons of overweight rats can be regarded as expression of neonatally programmed neurochemical plasticity of the regulatory system of body weight.

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.

Altered responses to orexigenic (AGRP, MCH) and anorexigenic (alpha-MSH, CART) neuropeptides of paraventricular hypothalamic neurons in early postnatally overfed rats.

Food intake and energy expenditure are regulated by neuropeptides in the hypothalamus. While cocaine- and amphetamine-regulated transcript (CART) peptide and melanocortins such as alpha-melanocyte-stimulating hormone (alpha-MSH) are anorexigenic and increase energy expenditure, the endogenous melanocortin receptor antagonist agouti gene-related protein (AGRP), melanin-concentrating hormone (MCH) and neuropeptide Y (NPY) are orexigenic, anabolic peptides. Alterations in the regulatory balance may promote excessive weight gain. The action of these peptides on paraventricular hypothalamic neurons was studied in brain slices of overweight, adult rats previously subjected to early postnatal overfeeding in small litters of only three pups per mother, compared to 12 pups per dam in control litters. CART, melanocortins and NPY significantly excited paraventricular neurons of controls, whereas neurons of small-litter rats were mainly inhibited. Inhibition was dominant following administration of AGRP, MCH and NPY. The altered responses of paraventricular neurons in adult small-litter rats might reflect a general mechanism of neurochemical plasticity and 'malprogramming' of hypothalamic neuropeptidergic systems acquired during the postnatal critical differentiation period, thus leading to permanently altered function of these regulatory systems of body weight.

Differential response to NPY of PVH and dopamine-responsive VMH neurons in overweight rats.

Neuronal responses to neuropeptide Y and dopamine were recorded in brain slices of hypothalamic paraventricular (PVH) and ventromedial (VMH) nuclei in normal and hyperphagic overweight rats reared in small litters of three pups. NPY significantly activated PVH neurons of normal rats, but inhibited neurons of overweight rats. In the VMH, a significantly higher coincidence of inhibition induced by NPY and dopamine was found in overweight rats. Similar neuronal responses were evoked by a NPY Y5 receptor agonist. Effects of NPY could be blocked by a Y1 receptor antagonist. The altered response of PVH neurons to the feeding-inducing NPY and the increased inhibition by NPY and dopamine in the VMH might contribute to the persisting hyperphagia and overweight of postnatally overnourished rats.

Hypothalamic ventromedial and arcuate neurons of normal and postnatally overnourished rats differ in their responses to melanin-concentrating hormone.

Melanin-concentrating hormone (MCH) is a neuropeptide involved in regulation of food intake and body weight. The study aimed to detect possible differences in responses of hypothalamic ventromedial and arcuate neurons to MCH, depending on the short-term nutritional state (fed versus food-deprived) and on the long-term state in overweight rats due to early postnatal overnutrition. The effect of MCH on a single-unit activity was studied in brain slices of normal and overweight rats. The latter (n=16) were raised till weaning in small litters (SL) of 3 pups compared to 10 pups in control litters (CL) and gained significantly greater body mass. Whereas MCH in effective concentrations in the pico- to nanomolar range could increase or suppress the activity of ventromedial or arcuate neurons studied in male normal fed or food-deprived (24 h) rats, its action became shaped in an unidirectional way in overweight, hyperphagic rats. Medial arcuate neurons (n=25) from hyperphagic rats were predominantly activated by MCH (p<0.05, paired t-test). This effect differed significantly from that induced on neurons (n=27) of control rats. Ventromedial neurons (n=34) of overweight rats were predominantly inhibited. Activation of arcuate neurons may induce feeding in particular through release of neuropeptide Y (NPY). Inhibition of ventromedial neurons may contribute to reduced energy expenditure. The increased expression of one response type to MCH by a neuronal population in overweight, hyperphagic rats might reflect a general mechanism of neurochemical plasticity and also suggest a participation of the peptide in long-term regulation of food intake and body weight in this model of obesity.

Increased inhibition by agouti-related peptide of ventromedial hypothalamic neurons in rats overweight due to early postnatal overfeeding.

Melanocortins, e.g. alpha-melanocyte stimulating hormone, are involved in the central nervous regulation of body weight. Agouti-related peptide (AGRP) as an endogenous melanocortin receptor antagonist induces feeding. Overexpression leads to obesity. Rats that grow up in small litters develop persistent hyperphagia and are overweight throughout life. Changes in the neuronal activity of the ventromedial hypothalamic nucleus (VMH) that signals satiety with activation might be involved. We studied single unit activity in hypothalamic brain slices. Melanocortins activated or inhibited similarly in control and small litter rats. AGRP mainly inhibited VMH neurons of overweight rats (t-test, P < 0.005, n = 33), whereas it also activated neurons of controls. This increased inhibition of VMH neurons by AGRP in early postnatally overnourished rats might contribute to the changed regulation leading to a persistent overweight condition throughout life.

Differential involvement of dopamine D1 and D2 receptors and inhibition by dopamine of hypothalamic VMN neurons in early postnatally overfed juvenile rats.

Dopamine is among the neurotransmitters involved in central regulation of food intake, and body weight control. To study possible changes in neuronal responses to dopamine, single unit activity of the ventromedial hypothalamic nucleus (VMN) was recorded in brain slices of normal and obese rats. The latter had developed overweight throughout juvenile life (p < 0.05) by early postnatal over-nourishment due to a reduction of litter size from 3rd to 21st day of life (small litters, SL). With effective concentrations of about 100-500 nM/I dopamine inhibited significantly more VMN neurons in obese than normal rats (Chi-square p < 0.05). While D2 receptors in the VMN are reported to mediate inhibition of food intake, the responses to dopamine were blocked by D2 receptor antagonists in significantly fewer neurons of SL than normal rats (p < 0.05). Furthemore, including results of action of D1 receptor agonists we found that significantly more neurons in SL than NL rats seem to express D1 receptors. Thus, increased suppression by dopamine of firing of VMN neurons that signal satiety with a rise in the discharge rate, and changed expression or activity of dopamine receptors might contribute to increased feeding behavior in juvenile rats hyperphagic and overweight due to early postnatal overfeeding.