Meena Jhanwar-Uniyal

Rapid and localized alterations of neuropeptide Y in discrete hypothalamic nuclei with feeding status

Neuropeptide Y (NPY) is believed to regulate the normal eating behavior and body weight in rats via central mechanisms. We have investigated whether NPY, which stimulates food intake, may in turn be modified by the nutritional state of the animals. Thus the impact of food deprivation (FD) (48 h) and subsequent refeeding on the levels of NPY in discrete hypothalamic areas was examined in this study. The results showed site specific change in only 3 of 7 hypothalamic sites. A 5-fold increment in NPY was reported in the paraventricular nucleus (PVN) and a 10-fold increase was observed in the arcuate nucleus-median eminence (ARC-ME). While subsequent refeeding for 6 h reversed the effect of FD in the ARC-ME, the levels of NPY in the PVN remained high in the refed rats. The perifornical lateral hypothalamus displayed a different pattern, namely, a significant increase in NPY content in refed as compared to satiated and deprived rats. The NPY levels in 4 other hypothalamic sites, namely, the dorsomedian, ventromedian, supraoptic and suprachiasmatic nuclei, and two extrahypothalamic sites, namely caudate nucleus and nucleus accumbens, showed total resistance to any change following deprivation and refeeding. These data emphasize the important and specific role of the paraventricular and arcuate nuclei in NPYs regulation of food intake and provide support for the idea that the variations of hypothalamic NPY after food deprivation reflect a specific physiological response of feeding regulatory system to alterations in the animal nutritional state and body weight.

Distribution of α-adrenergic, β-adrenergic and dopaminergic receptors in discrete hypothalamic areas of rat

Summary Catecholamine receptor binding sites were measured in discrete hypothalamic nuclei or regions as well as in certain extrahypothalamic areas of the adult male rat. For each assay, discrete areas were microdissected from frozen tissue sections and pooled from several animals. Specific high affinity binding sites were assessed at fixed ligand concentrations for [ 3 H] p -aminoclonidine (PAC) and [ 3 H](2-C 2′,6′-(CH 3 O) 2 phenoxyethylamino)-methylbenzodioxan (WB-4101) for α-adrenergic receptor sites, for [ 3 H]dihydroalprenolol (DHA) for β-adrenergic receptor sites, and for [ 3 H]2-amino-6, 7-dihydroxy-1,2,3,4-tetrahydronaphtalene (ADTN) and [ 3 H]spiroperidol in the presence of cinanserin for dopaminergic receptor sites. Regional variations in [ 3 H]WB-4101 binding were relatively small in magnitude, with most hypothalamic and extrahypothalamic areas possessing between 60 and 90% of the binding in frontal cortex. [ 3 H]PAC binding showed a wider range of binding density across brain areas than did [ 3 H]WB-4101, but, in general, variations in [ 3 H]PAC binding paralleled those in [ 3 H]WB-4101 binding. In hypothalamus, binding was characterized as being predominantly to α 1 -receptors in the of [ 3 H]WB-4101 and to α 2 -receptors in the case of [ 3 H]PAC. The medial hypothalamic areas exhibited a somewhat higher density of these α-adrenergic sites than did the lateral hypothalamus (perifornical hypothalamus and medial forebrain bundle). Also, the ratio of [ 3 H]PAC to [ 3 H]WB-4101 binding differed in different hypothalamic areas, ranging from 1.5:1 to 4:1. The median eminence was exceptional in that it contained appreciable [ 3 H]PAC but no significant [ 3 H]WB-4101 binding sites at the radioligand concentrations tested. Binding of [ 3 H]DHA to β-adrenergic receptors varied over approximately a 3-fold range in the different hypothalamic areas, with binding highest in the medial forebrain bundle and the medial preoptic area, and lowest in the periventricular, dorsomedial and posterior hypothalamic nuclei, the median eminence and the zona incerta. The ratio of β-adrenergic to α-adrenergic binding sites was generally lower in the medial than in the lateral hypothalamic areas and higher in the extrahypothalamic areas examined than in the hypothalamus. With regard to [ 3 H]spiroperidol and [ 3 H]ADTN binding to dopaminergic sites, the striatum, nucleus accumbens and olfactory tubercle showed a greater density of [ 3 H]spiroperidol than of [ 3 H]ADTN sites, in contrast to the hypothalamus where [ 3 H]ADTN binding was more predominant. Within the hypothalamus, [ 3 H]ADTN binding was relatively uniform, while [ 3 H]spiroperidol binding was quite high in four hypothalamic areas (lateral perifornical area, medial forebrain bundle, paraventricular and dorsomedial nuclei), intermediate in the median eminence and arcuate nucleus, and low or not detectable in all other hypothalamic areas.

Diurnal rhythm of neuropeptide Y-like immunoreactivity in the suprachiasmatic, arcuate and paraventricular nuclei and other hypothalamic sites

The diurnal rhythm of neuropeptide Y (NPY)-like immunoreactivity was examined in 9 discrete hypothalamic sites of rats maintained on a 12:12 h light/dark cycle. Significant bimodal rhythms of NPY concentration were detected in the suprachiasmatic and arcuate nuclei, with significant peaks just prior to onset of the nocturnal period and also at onset of the light period. In the parvocellular division of the paraventricular nucleus, a unimodal NPY peak was observed prior to dark onset. No diurnal rhythm was seen in the magnocellular division of the paraventricular nucleus, nor in 5 other hypothalamic areas examined.

Neuropeptide Y projection from arcuate nucleus to parvocellular division of paraventricular nucleus : specific relation to the ingestion of carbohydrate

Neuropeptide Y (NPY) injection into the hypothalamic paraventricular nucleus (PVN) stimulates feeding behavior and specifically carbohydrate intake in rats. The present study investigated the relation between endogenous levels of NPY and natural ingestion for carbohydrate. It also examined the possible importance of a specific NPY projection in this relationship, which traverses from cell bodies in the arcuate nucleus (ARC) to terminals in the parvocellular division of the PVN (pPVN). Sprague-Dawley rats were given pure macronutrient diets (carbohydrate, protein and fat), and their daily nutrient intake was recorded for 3 weeks. The rats were sacrificed, and 8 hypothalamic nuclei were micropunched and examined via RIA for endogenous NPY levels. The results demonstrate a strong, positive correlation between daily carbohydrate intake and hypothalamic NPY levels. The relationship was specific to the pPVN (r = +0.71; P < 0.001), ARC (r = +0.57; P < 0.001) and dorsomedial nucleus (DMN, r = +0.52; P < 0.01), and was not observed in any other hypothalamic area, including the magnocellular division of the PVN. In the pPVN, the NPY levels of animals that consumed > 50 kcal of carbohydrate (49 pg/microgram protein) were almost twice that of animals that consumed < 20 kcal of carbohydrate (28 pg/microgram protein: P < 0.01). Furthermore, NPY levels in the ARC were positively correlated with NPY in the pPVN and DMN but not any other nuclei. No relation between hypothalamic NPY and measures of protein or fat ingestion was detected. Levels of NPY were also unrelated to total caloric intake, to body weight at sacrifice, and to body weight gain during the 3-week measurement period. These results, together with other findings, provide support for a role of endogenous NPY and its projection from the ARC to the pPVN, perhaps via the DMN, in controlling natural appetite for carbohydrate.

Diurnal rhythm of α2-noradrenergic receptors in the paraventricular nucleus and other brain areas: Relation to circulating corticosterone and feeding behavior

The paraventricular nucleus alpha 2-noradrenergic system and the glucocorticoid hormone, corticosterone, are known to modulate feeding behavior and exhibit a circadian pattern which may be related to the natural periodicity of feeding in the rat. The results of the present study indicate that the binding of [3H]p-aminoclonidine to alpha 2-noradrenergic receptors specifically in the paraventricular nucleus varies concomitantly with plasma corticosterone levels, as well as spontaneous feeding. A monophasic peak of paraventricular noradrenergic receptor binding is detected at the onset of the dark period, when corticosterone levels are highest and feeding is initiated. On the other hand, the supraoptic nucleus exhibits the reverse diurnal pattern, i.e., a significant decline of [3H]p-aminoclonidine binding at the onset of the dark period. Other hypothalamic and extra-hypothalamic areas fail to show significant changes in alpha 2-noradrenergic receptors as a function of the diurnal cycle. This study supports other evidence indicating a close interaction between circulating corticosterone and alpha 2-noradrenergic receptors in specific hypothalamic areas. It also reveals a potential importance for this interaction in control of the natural feeding rhythm.

Impact of circulating corticosterone on α1- and α2-noradrenergic receptors in discrete brain areas

The impact of adrenalectomy (ADX) and subsequent corticosterone (CORT) replacement, on the binding of [3H]p-aminoclonidine to α2-noradrenergic receptors and [3H]prazosin to α1-noradrenergic receptors, was studied in 8 discrete hypothalamic and 5 extra-hypothalamic areas of rats. With little change in extra-hypothalamic receptors, ADX produced a large CORT-reversible decrease in α2-receptor binding specifically within the hypothalamic paraventricular nucleus (PVN) and a CORT-reversible increase within the supraoptic nucleus. α1-Noradrenergic receptors, in contrast, were generally unaffected by ADX. This and other evidence leads us to propose a potential modulatory influence of circulating CORT on hypothalamic α2 receptors and a specific function for this CORT-α2 receptor interaction specifically within the PVN, in the control of eating behavior.

ANORECTIC EFFECT OF METFORMIN IN OBESE ZUCKER RATS : LACK OF EVIDENCE FOR THE INVOLVEMENT OF NEUROPEPTIDE Y

The hypothalamic neuropeptide Y content and preproneuropeptide Y mRNA expression were studied in metformin-treated (300 mg/kg orally for 12 days), in pair-fed and in ad libitum-fed obese Zucker rats in order to elucidate possible mechanisms involved in the anorectic and body weight reducing effect of chronic metformin treatment in genetically obese Zucker rats. In addition the acute influence of metformin on food intake was studied by comparing its effects after oral and parenteral administration. The concentration of neuropeptide Y in the hypothalamic paraventricular nucleus was significantly higher in the metformin-treated and pair-fed rats when compared to the control animals. The expression of preproneuropeptide Y mRNA in the arcuate nucleus was similar in all three treatment groups. Both chronic metformin treatment and pair-feeding markedly lowered hyperinsulinaemia in these animals. A single subcutaneous dose of metformin (300 mg/kg) reduced food intake only in obese animals, while the same dose of metformin given orally did not affect food intake in either lean or obese animals. It is concluded that the treatment with metformin and pair-feeding, which results in comparable reductions in food intake, body weight gain and hyperinsulinaemia, similarly increase neuropeptide Y concentrations in the paraventricular nucleus while not affecting preproneuropeptide Y mRNA expression in the arcuate nucleus. The increase in hypothalamic neuropeptide Y content may be secondary to the reduction in hyperinsulinaemia during metformin treatment and pair-feeding. Thus, the anorectic effect of chronic metformin treatment cannot be explained by changes in content or expression of hypothalamic neuropeptide Y.

Destruction of noradrenergic innervation to the paraventricular nucleus: deficits in food intake, macronutrient selection, and compensatory eating after food deprivation

Norepinephrine (NE) injected into the paraventricular nucleus (PVN) has a stimulatory effect on feeding behavior and is found to selectively enhance preference for carbohydrate in the rat. The present experiments were conducted to assess the impact of chronic depletion of NE within the PVN on food intake and appetite regulation. The catecholamine (CA) neurotoxin, 6-hydroxydopamine (6-OHDA), when administered into the PVN, produced a significant depletion of PVN NE in association with a variety of behavioral changes. The immediate consequence of the neurotoxin lesion was a dramatic increase in 24-hr food intake, attributed predominantly to a preferential increase in carbohydrate and fat consumption. The long-term effects related to CA depletion were a deficit in daily food consumption, particularly of carbohydrate (-42%). Although animals with diminished PVN NE maintained a normal diurnal feeding pattern, they failed to exhibit the increased ingestion of an energy-rich carbohydrate diet which rats normally show during the dark period of the diurnal cycle. Rats injected with 6-OHDA directly into the PVN exhibited a normal response to glucoprivic challenge, but demonstrated a deficit in their ability to produce compensatory feeding, particularly of carbohydrate and fat, in response to food deprivation. These findings suggest a specific function for PVN noradrenergic mechanisms in normal energy repletion when body energy stores are reduced.

Critical effects of aging and nutritional state on hypothalamic neuropeptide Y and galanin gene expression in lean and genetically obese Zucker rats

We have investigated, by Northern blot analysis, the hypothalamic gene expression [messenger RNA (mRNA)] of two appetite stimulating neuropeptides, neuropeptide Y (NPY) and galanin (GAL) in lean (+/+) and genetically obese (fa/fa) Zucker rats at 11, 24 and 40 weeks of age and their responsiveness to food deprivation. At 11 weeks of age, hypothalamic NPY mRNA levels of fa/fa rats were similar to those observed in lean littermates. However, NPY mRNA levels of fa/fa rats were significantly greater than those of lean rats at 24 (+126%; P < 0.01) and 40 (+65%; P < 0.05) weeks of age. Food deprivation caused a significant increase in NPY mRNA levels in both lean and fa/fa Zucker rats at 11 and 24 weeks of age, but not at 40 weeks old rats. Hypothalamic GAL mRNA showed a different pattern of change. The relative content of GAL mRNA in 11 week old obese rats was significantly lower (-68%; P < 0.05) than that of lean rats, while GAL mRNA was significantly higher in 40 week old (+57%; P < 0.05) obese rats compared to their lean littermates. At 24 weeks of age, hypothalamic GAL mRNA levels did not differ between lean and obese rats. Food deprivation induced no change in hypothalamic GAL mRNA in lean rats of all 3 ages; however, it caused an increase of GAL mRNA in obese rats at 11 (+60%; P < 0.05) and 24 (+44%; P < 0.05) weeks, but not at 40 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)

Alterations in catecholamine levels and turnover in discrete brain areas after food deprivation

The present study determined the levels and turnover of norepinephrine (NE), epinephrine (EPI) and dopamine (DA) in discrete brain areas of rats after 48 hr food deprivation. The steady-state levels of NE, EPI and DA in saline-treated food-deprived rats, relative to satiated rats, remained basically unchanged. However, 48 hr deprivation caused a site-selective potentiation, specifically in the hypothalamic paraventricular nucleus, in the depletion of NE after alpha-methyl-p-tyrosine injection (IP, 200 mg/kg), indicating an increase in NE turnover. While changes in EPI turnover could not be demonstrated, an apparent increase in DA turnover was detected in the perifornical lateral hypothalamus and anterior hypothalamic nucleus after deprivation, while decreased DA turnover was seen in the hypothalamic dorsomedial nucleus and caudate nucleus. These results may reflect specific functions of hypothalamic catecholamines in control of food intake.