Insulin and the paraventricular hypothalamus: modulation of energy balance

The effects of insulin injections (0.1, 1, 10 and 40 mU) into the paraventricular hypothalamus (PVN) were investigated in an open-circuit calorimeter. Wistar rats were tested, with no food available during the tests. The 0.1 and 1 mU doses had no effects on either respiratory quotient or energy expenditure. The 10 mU dose increased respiratory quotient which indicates increased dependency on carbohydrates as an energy substrate. The same dose had no effects on thermogenesis. In contrast, the 40 mU dose decreased respiratory quotient which indicates increased dependency on fats as an energy substrate. The higher dose also increased thermogenesis. Since neither dose significantly affected locomotor activity, the metabolic data are not confounded with activity effects. These data indicate that insulin in the PVN produces a primary modulation of the metabolic parameters central to maintaining energy balance. In separate experiments, the 4 doses of insulin reduced food intake and body weight over a 24 h period. They also produced a dose-related increase in blood glucose concentration over a one hour period. Taken together, these findings are interpreted in a model in which insulin in the PVN acts as a signal indicating increased body fat. This increases thermogenesis, fat utilization and glycemic levels, and inhibits feeding. The net effect of this integrated metabolic-behavioural response is a regulatory reduction in body fat.     

Glucagon and the paraventricular hypothalamus: modulation of energy balance

The effects of glucagon injections (25, 100 and 200 ng) into the paraventricular nucleus of the hypothalamus (PVN) were investigated in an open-circuit calorimeter. Wistar rats were tested, with no food available during the tests. High doses of glucagon (100 and 200 ng) produced small and short-lasting increases in energy expenditure. The independence of these changes from changes in locomotor activity suggests that the thermogenesis represents a primary modulation and is not secondary to increased locomotion. All three doses of glucagon produced long-lasting and dose-related increases in respiratory quotient which were unrelated to any changes in locomotor activity. As with the changes in energy expenditure, this dissociation indicates that the effects are not secondary to changes in locomotor activity. These data constitute the first evidence that glucagon in the PVN modulates the metabolic parameters central to energy balance. In separate experiments, the three doses of glucagon increased blood glucose concentration over a one hour period, but they did not affect food and water intake and body weight over 24 h. These findings suggest that glucagon normally acts on the PVN in conditions of increased body fat to initiate autonomic mechanisms which increase glycemic levels, thermogenesis and carbohydrate utilization. These data constitute the first direct evidence for the involvement of the PVN in the regulation of energy balance by glucagon. The main effect of glucagon in the PVN is anabolic in that it increases dependence on carbohydrates as an energy substrate which results in a sparing of fat reserves.     

Effect of central administration of QRFP(26) peptide on energy balance and characterization of a second QRFP receptor in rat

The recently identified neuropeptide QRFP(26) is predominantly expressed in the hypothalamus and was suggested to play a role in the regulation of food intake following the observation of an acute orexigenic effect after central administration in mice. QRFP(26) exerts its effect via GPR103 and a newly identified receptor in mouse. The aim of our study was (a) to investigate the distribution of QRFP(26) and a newly discovered QRFP receptor mRNA in rat and (b) to further characterize the effects of central administration of QRFP(26) on energy balance in rats. QRFP(26) mRNA was detected in the retrochiasmatic nucleus, periventricular nucleus, arcuate nucleus and restricted areas of the lateral nucleus of the hypothalamus. We found an additional receptor with high homology for GPR103 in rat. This receptor increases inositol triphosphate production in transfected cells in presence of QRFP(26) and its mRNA was particularly enriched in ventral and posterior thalamic groups, anterior hypothalamus and medulla. When QRFP(26) (10 microg and 50 microg) was administered centrally before the start of the light phase both doses increased food intake for 2 h after injection without reaching statistical significance. QRFP(26) caused no changes in locomotor activity or energy expenditure. In summary, central QRFP(26) injection causes slight and transient hyperphagia in rats without changing any other energy balance parameters after 24 h. We conclude that QRFP(26) has limited impact on the central regulation of energy balance in rats and that its essential function remains to be clarified.     

Cognitive dysfunction, urinary retention, and a lesion in the thalamus--beware of possible toxocariasis of the central nervous system

Toxocariasis of the central nervous system is usually characterized by an eosinophilic meningitis, encephalitis or myelitis. We here report a patient with an at least 7 years history of unexplained neurologic signs and symptoms. MRI showed a cystic lesion in the left thalamus, compatible with a parasitic infection. Blood and CSF analyses were positive for Toxocara canis IgG Western-blot, but were otherwise unremarkable. The case report raises the question whether there are chronic or late variants of this disease.     

Somatostatin and the paraventricular hypothalamus: modulation of energy balance

The effects of somatostatin injections (0.1, 1 and 5 μg) into the paraventricular nucleus of the hypothalamus (PVN) were investigated in an open-circuit calorimeter. Wistar rats were tested, with no food available during the tests. The 0.1 and 1 μg doses produced large and long-lasting decreases in respiratory quotient, which indicates the preferential utilization of fats as an energy substrate. The 5 μg dose produced a brief decrease in energy expenditure. Locomotor activity was not affected by any treatment which indicates that the effects on respiratory quotient and energy expenditure are not secondary to changes in activity. These findings demonstrate that somatostatin in the PVN inhibits thermogenesis and induces the preferential utilization of fats while sparing carbohydrate reserves. However, it is significant that the effects on energy expenditure and energy substrate utilisation occured at different doses. These data constitute the first evidence that somatostatin in the PVN produces a primary modulation of the metabolic parameters central to energy balance. In separate experiments, all three doses of somatostatin increased blood glucose concentration over a one hour period, and the 5 μg dose decreased body weight over a 24 h period. Food and water intake were not affected by the somatostatin injections. Taken together, these findings are interpreted in a model in which somatostatin is a signal to the PVN of increased body fat. This mobilises sympathetic mechanisms which increase fat utilization and blood glucose levels.     

The effects of galanin-like peptide on energy balance, body temperature and brain activity in the mouse and rat are independent of the GALR2/3 receptor

Galanin-like peptide (GALP) is a neuropeptide that is thought to act on the galanin receptors GALR1, GALR2 and GALR3. In rats, i.c.v. injection of GALP has dichotomous actions on energy balance, stimulating feeding over the first hour, but reducing food intake and body weight at 24 h, as well as causing an increase in core body temperature. In mice, GALP only induces an anorexic action, and its effects on core body temperature are unknown. One aim of the present study was to determine the effects of GALP on core body temperature in mice. Intracerebroventricular injection of GALP into conscious mice had no effect on feeding over 1 h, but caused a significant reduction in food intake and body weight at 24 h. It also caused an immediate drop in core body temperature, which was followed by an increase in body temperature. To understand these different effects of GALP on energy balance in mice compared to rats, and to determine the involvement of GALR2 and GALR3, immunohistochemistry was performed to localise c-Fos, a marker of cell activation. Intracerebroventricular injection of GALP induced c-Fos expression in the parenchyma surrounding the ventricles, the ventricular ependymal cells and the meninges in mice and rats. GALP also induced c-Fos expression in the supraoptic nucleus, dorsomedial hypothalamic nucleus, lateral hypothalamus and nucleus tractus solitarius in rats but not in mice. Central administration of a GALR2/3 agonist in rats did not induce c-Fos in any of the brain regions that expressed this protein after GALP injection, and had no effect on food intake, body weight and body temperature in rats or mice. These data suggest that GALP induces differential effects on energy balance and brain activity in mice compared to rats, which are unlikely to be due to activation of the GALR2 or GALR3 receptor.     

Tumor necrosis factor and interleukin-1β: suppression of food intake by direct action in the central nervous system

Intracerebroventricular microinfusion of recombinant human tumor necrosis factor (rhTNF) and recombinant human interleukin-1β (rhIL-1β) suppressed food intake in rats. Central infusion of heat-inactivated rhTNF and rhIL-1β, bovine serum albumin, heparin or transforming growth factor-β had no such effect. Central infusion of rhIL-1β did not affect the dipsogenic response to central administration of angiotensin II. Peripheral administration of rhTNF and rhIL-1β in doses equivalent to or higher than those administered centrally had no effect. Electrophoretically applied rhTNF and rhIL-1β specifically suppressed the activity of glucose-sensitive neurons in the lateral hypothalamic area. Glucose-insensitive neurons were little affected. The results suggest that TNF and IL-1β act directly in the central nervous system to suppress feeding, and this effect may be operative during acute and chronic disease.     

Differential effects of serotonin (5-HT) lesions and synthesis blockade on neuropeptide-Y immunoreactivity and 5-HT1A, 5-HT1B/1D and 5-HT2A/2C receptor binding sites in the rat cerebral cortex

The present study was aimed at comparing the effects of serotonin (5-HT) synthesis blockade using chronic administration of p-chlorophenylalanine (PCPA) and 5,7-dihydroxytryptamine injections of variable volume (3 vs. 6 μl) on the density of NPY immunoreactive (Ir) neurons and binding of [³H]8-OH-DPAT, S-CM-G[¹²⁵I]TNH₂ and [¹²⁵I]DOI to 5-HT_1A, 5-HT_1B/1D, and 5-HT_2A/2C receptors in rat cortical regions. Three weeks after large but partial (89% depletion in 5-HT tissue concentration) lesions of 5-HT neurons no changes in neither NPY immunoreactivity nor 5-HT receptor binding were detected. The complete 5,7-DHT lesions produced increases in the number of NPY-Ir neurons in the upper regions of the cingular (134%), frontal (140%) and parietal cortex (48%) and corresponding decreases in 5-HT_2A/2C binding (16–26%). No changes in 5-HT_1A and 5-HT_1B/1D binding were observed after lesions of this kind. After PCPA treatment, decreases in NPY-Ir neurons density (22–40%) and increases in 5-HT_1A and 5-HT_1B/1D receptor binding sites (20–50%) were distributed in both upper and deeper cortical regions. The lack of effect of the partial lesion suggests that spared 5-HT neurons may exert compensatory mechanisms up to a large extent. The changes in NPY immunoreactivity and 5-HT_2A/2C binding detected in the upper regions of the cortex after complete 5-HT lesions probably result from local cellular rearrangements, whereas blocking 5-HT synthesis has more widespread influence on NPY neurons and on 5-HT_1A and 5-HT_1B/1D receptor subtypes. Moreover, decreases in DOPAC concentrations detected only after complete lesions suggest that the involvement of catecholaminergic transmission may also differentiate 5,7-DHT and PCPA treatments. Altogether, these data suggest that different receptor subtypes might be involved in 5-HT–NPY relationships.     

Developmental changes in the expression of GABAA receptor subunits alpha1, alpha2, and alpha3 in brain stem nuclei of rats

Gamma-aminobutyric acid (GABA)(A) receptor subunit switching is a suggested postnatal mechanism for changes in GABA transmission from depolarization to hyperpolarization. Previously, we found an apparent switch between GABA(A) alpha3 and alpha1 subunit expression in the rat pre-B?tzinger complex (PBC) on postnatal day (P) 12, a presumed peak critical period of respiratory nuclei development. The present study aimed at determining if GABA(A) subunit switching occurred in another respiratory nucleus, the ventrolateral subnucleus of the solitary tract nucleus (NTS(VL)), and in a non-respiratory cuneate nucleus (CN) of P0 to P21 rats. In both nuclei: (1) the expression of GABA(A) alpha1 subunit was relatively low at birth but increased with development; (2) that of GABA(A) alpha3 was relatively high at birth but declined with age; and (3) GABA(A) alpha2 remained relatively low and constant throughout development. However, the specific patterns differed between the two nuclei, but were similar between the NTS(VL) and the PBC. In the NTS(VL), GABA(A) alpha1 expression gradually increased from birth and peaked at P12, whereas that in the CN sharply rose from P7 and peaked at P10. GABA(A) alpha3 expression had a prominent decrease from P11 to P12 in the NTS(VL), whereas that in the CN only gradually declined from P10 to P12. The developmental trends of alpha1 and alpha3 in the NTS(VL) intersected close to P12, whereas those in the CN intersected at P10. Despite differences in timing, GABA(A) alpha subunit switching may be a common theme in the brain stem that may mediate different functional properties of GABA transmission.     

From antibody to clinical phenotype,the black box of the antiphospholipid syndrome: Pathogenic mechanisms of the antiphospholipid syndrome

The antiphospholipid syndrome (APS) is diagnosed by the combination of vascular thrombosis and/or pregnancy morbidity and the detection of antiphospholipid antibodies (aPLs) in plasma. In the last few years, a great effort has been made to unravel the mechanism by which aPLs cause thrombosis and a vast amount of mechanisms have been proposed. aPLs were proposed to induce a prothrombotic state by influencing the cellular blood compartment, the plasma compartment, the vascular wall and even metabolic pathways beyond the hemostatic system. However, due to the diversity in the mechanisms and the differences in the methodology, the focus of the mechanistical studies in this field seems to be largely diffused. It is hard to imagine that aPLs can exert such a diversity of effects, resulting in either thrombosis and/or pregnancy morbidity and the relationship between aPLs and the clinical manifestations remains to be a mysterious “black box”. In an attempt to get insight in what takes place inside the black box, we have analyzed 126 mechanistical studies on aPLs and discussed differences in the type of antibodies that were used, the involvement of beta2-glycoprotein I (β₂GPI), and the criteria used to diagnose APS patients.     

Defining Desirable Central Nervous System Drug Space through the Alignment of Molecular Properties, in Vitro ADME, and Safety Attributes

As part of our effort to increase survival of drug candidates and to move our medicinal chemistry design to higher probability space for success in the Neuroscience therapeutic area, we embarked on a detailed study of the property space for a collection of central nervous system (CNS) molecules. We carried out a thorough analysis of properties for 119 marketed CNS drugs and a set of 108 Pfizer CNS candidates. In particular, we focused on understanding the relationships between physicochemical properties, in vitro ADME (absorption, distribution, metabolism, and elimination) attributes, primary pharmacology binding efficiencies, and in vitro safety data for these two sets of compounds. This scholarship provides guidance for the design of CNS molecules in a property space with increased probability of success and may lead to the identification of druglike candidates with favorable safety profiles that can successfully test hypotheses in the clinic.