E. De Castro e Silva

Hyperglycemia induced by acute central fluoxetine administration: role of the central CRH system and 5-HT3 receptors

Brain serotonin and CRH systems participate in the control of blood glucose levels. We have previously demonstrated that the pharmacological stimulation of central 5-HT3 receptors, the target for several therapeutic agents used as antiemetics in the course of chemotherapy, induces hyperglycemia. The aim of the present study was to investigate the participation of the brain CRH component and 5-HT3 receptors in basal blood glucose levels as well as in the hyperglycemia induced by third ventricle injections of fluoxetine, a serotonin reuptake inhibitor with a broad range of clinical use. In this study, we used fasted adult Wistar male rats (220 +/- 20 g) whose third ventricles were cannulated 7 days prior to the experiments. Acute third ventricle injections of fluoxetine caused a significant increase in plasma glucose levels throughout the experiment. Pretreatment with alpha-helical CRH, a selective CRH antagonist, significantly blunted fluoxetine-induced hyperglycemia. Also, pretreatment with two distinct selective 5-HT3 receptor antagonists (LY-278,584 and ondansetron) significantly impaired the rise in plasma glucose levels observed in fluoxetine-treated animals pretreated with isotonic saline solution. None of these antagonists was able to modify blood glucose levels when injected alone into the third ventricle. Animals receiving third ventricle injections of fluoxetine, in spite of being hyperglycemic, presented plasma insulin levels similar to those displayed by normoglycemic, saline-treated controls. It is suggested that the acute increase in brain serotonergic activity caused by third ventricle injections of fluoxetine induces a hyperglycemic response that requires the functional integrity of the brain CRH system and 5-HT3 receptors. Also, it is proposed that the absence of a compensatory increase in plasma insulin levels may contribute to the generation of a hyperglycemic response after central fluoxetine administration.

Effect of central acute administration of cadmium on drinking behavior.

The effect of acute third ventricle cadmium administration on the drinking behavior of adult male rats under different situations was studied. Injections of cadmium chloride (0.07, 0.7,and 7.0 ng/rat) significantly attenuated water intake in dehydrated rats. Drinking behavior induced by acute intracerebroventricular injections of carbachol (2 micrograms/rat) or angiotensin II (5 ng/rat) was also inhibited by central cadmium injections. Cadmium-induced blockade in water intake in dehydrated animals was reverted by the previous administration of a 5-HT2 antagonist (RP62203) in different doses (5 and 10 micrograms/rat). The data clearly reveal that cadmium elicits very fast actions on the central nervous system. It is suggested that cadmium-induced attenuation of water intake may rely on at least three different mechanisms: impairment of cholinergic and angiotensinergic systems in the brain and stimulation of a central serotonergic drive acting on 5-HT2 receptors. The study of cadmium neurotoxicity by observation of drinking behavior, a behavioral parameter easy to be recorded and measured, is proposed.

Central administration of mCPP, a serotonin 5-HT2B/2C agonist, decreases water intake in rats

In the present study, we investigated in rats the effect of third ventricle injections of 1-(3-chlorophenyl)piperazine (mCPP), a 5-HT(2) receptor agonist, on water intake induced by three different physiological stimuli: fluid deprivation, acute salt load and hypovolemia. Injections of mCPP in the doses of 80 and 160 nmol/rat were able to decrease water intake in all three conditions studied. Third ventricle injections of mCPP (160 nmol/rat) were no longer able to diminish water intake in the groups of rats pretreated with central injections of an equimolar amount of (+)-cis-4,5,7a,8,9,10,11,11a-octahydro-7H-10-methylindolo[1,7-bc][2,6]-naphthyridine (SDZ SER 082), a selective 5-HT(2B/2C) antagonist. The central administration of mCPP (160 nmol/rat) was not able to modify the intake of a 0.1% saccharin solution. It is suggested that the central activation of a 5-HT(2B/2C) component is able to impair the drive for water intake induced by the physiological stimuli represented by fluid deprivation, acute salt load and hypovolemia. This effect seems not to be consequent on a general nonspecific central nervous system depression or on a locomotor deficit, because saccharin intake is not affected by third ventricle injections of mCPP.

Acute fluoxetine administration differentially affects brain C-Fos expression in fasted and refed rats

In the present study we investigated the effect of acute fluoxetine administration on the expression of c-Fos in the rat brain under two different metabolic conditions: fed and fasting states. Wistar male rats, weighing 220+/-30g, received i.p. injections of saline solution or fluoxetine (10mg/kg), and were killed 2 h later. The brains were removed after transcardiac perfusion with phosphate-buffered saline followed by paraformaldehyde, and were then processed for immunohistochemistry. Fos-like immunoreactivity was quantified by a computerized system. Fasted animals faced an 18-h suppression of food intake, while fed groups were submitted to an initial 14-h period of fast followed by a 4-h period in which food was freely available. Both in fasting and fed states, fluoxetine-treated animals presented a significant increase in c-Fos expression in hypothalamic areas, limbic structures, circumventricular areas, and in mesencephalic and rhomboencephalic regions, as compared with saline-treated controls. The quantitative comparison of data obtained from fasted and fed animals showed that fasted rats treated with fluoxetine presented a higher c-Fos expression in the ventromedial hypothalamus and the paraventricular nuclei compared with the fed group, while in fluoxetine-treated fed rats c-Fos expression was higher in the arcuate nuclei, medial amygdala, locus coeruleus and dorsal raphe nuclei, as compared with fasted, fluoxetine-treated animals. These data indicate that the metabolic condition of the animals significantly modifies fluoxetine-induced brain c-Fos expression, suggesting that visceral and behavioral fluoxetine effects may be influenced by the metabolic state of the individual.

The central amygdala regulates sodium intake in sodium-depleted rats: Role of 5-HT3 and 5-HT2C receptors

In the present paper, we have evaluated the participation of 5-HT(3) and 5-HT(2C) receptors in the central amygdala (CeA) in the regulation of water and salt intake in sodium-depleted rats. m-CPBG-induced pharmacological activation of 5-HT(3) receptors located in the CeA resulted in a significant reduction in salt intake in sodium-depleted rats. This antinatriorexic effect of m-CPBG was reverted by pretreatment with the selective 5-HT(3) receptor antagonist ondansetron. The injection of ondansetron alone into the CeA had no effect on sodium-depleted and normonatremic rats. Conversely, pharmacological stimulation of 5-HT(2C) receptors located in the central amygdala by the selective 5-HT(2C) receptor agonist m-CPP failed to modify salt intake in sodium-depleted rats. Additionally, the administration of a selective 5-HT(2C) receptor blocker, SDZ SER 082, failed to modify salt intake in rats submitted to sodium depletion. These results lead to the conclusion that the pharmacological activation of 5-HT(3) receptors located within the CeA inhibits salt intake in sodium-depleted rats and that 5-HT(2C) receptors located within the CeA appear to be dissociated from the salt intake control mechanisms operating in the central amygdala.

Acute effect of intracerebroventricular administration of lead on the drinking behavior of rats induced by dehydration or central cholinergic and angiotensinergic stimulation

In the present paper, the acute effect of third ventricle injections of lead acetate (5, 10, 100, 1000 ng/rat) on the drinking behavior of adult, male, Wistar rats was investigated. Lead generates a prompt and significant reduction in water intake induced by three different circumstances: dehydration (14 h of water deprivation) and after carbachol (2 micrograms/rat, ICV) or angiotensin II (10 ng/rat, ICV) administration. These results show that lead may produce very fast actions in the central nervous system and suggest that inhibition of water intake by lead may depend on impairment of central cholinergic and/or angiotensinergic functions.

Multiple opioid receptors mediate the hypotensive response induced by central 5-HT3 receptor stimulation

The aim of the present work was to investigate the role of brain μ, κ and δ opioid receptors in the central serotonergic mechanisms regulating blood pressure in rats. The data obtained show that: (1) pharmacological activation of central 5-HT(3) receptors yields a significant decrease in blood pressure; (2) the blockade of those receptors by a selective antagonist induces an acute hypertensive response; (3) the pharmacological blockade of central opioid receptors by three different opioid antagonists exhibiting variable degrees of selectivity to μ, κ and δ opioid receptors always suppressed the hypotensive response induced by central 5-HT(3) receptor stimulation; (4) the blockade of opioid receptors by the same opioid antagonists that impaired the hypotensive effect of central 5-HT(3) receptor stimulation failed to modify blood pressure in animals not submitted to pharmacological manipulations of central 5-HT(3) receptor function. It is shown that a 5-HT(3) receptor-dependent mechanism seems to be part of the brain serotonergic system that contributes to cardiovascular regulation since the hypertensive response observed after ondansetron administration indicates that central 5-HT(3) receptors exert a tonic inhibitory drive on blood pressure. Furthermore, the data obtained here clearly indicate that the hypotensive response observed after pharmacological stimulation of central 5-HT(3) receptors depends on the functional integrity of brain μ, κ and δ opioid receptors, suggesting that a functional interaction between serotonergic and opiatergic pathways in the brain is part of the complex, multifactorial system that regulates blood pressure in the central nervous system.

Blockade of central kappa-opioid receptors inhibits the antidipsogenic effect of interleukin-1β

The objective of the present study was to investigate the role of brain kappa-opioid receptors (KOR) in the antidipsogenic effect promoted by third ventricle injections of interleukin-1beta (IL-1beta). Wistar male rats were submitted to three different, thirst-inducing, physiological conditions: dehydration induced by water deprivation, hyperosmolarity induced by salt-load and hypovolemia induced by polyethylene glycol subcutaneous injection. Third ventricle injections of IL-1beta significantly inhibited the increase in water intake observed in those situations. The pharmacological blockade of central KOR by the selective KOR antagonist nor-binaltorphimine (BNI) at different doses significantly inhibited the antidipsogenic effect induced by the central administration of IL-1beta in all conditions tested: dehydration, hypovolemia and hyperosmolarity. The central administration of IL-1beta failed to induce any locomotor deficit, as verified in an open field test. Stimulation of the central interleukinergic component did not result in any general suppression of ingestive behavior since no change in saccharin intake was recorded during a dessert test in animals receiving central injections of IL-1beta. Furthermore, the inhibitory effects of IL-1beta on water intake cannot be attributed to sickness-like effects induced by these compounds, since an aversion test excluded this possibility. In summary, the data shown in the present study clearly show that the antidipsogenic effect observed in rats following third ventricle injections of IL-1beta depend on the functional integrity of a brain kappa-opioid-dependent component.

c-Fos expression identifies brain areas activated in response to nortriptyline.

1 Roth B, Sheffler DJ, Kroeze WK. Nat Rev Drug Discov 2004; 3: 353–359. 2 Homayoun H, Moghaddam B. Biol Psychiatry 2007; 61: 679–687. 3 Leveque J-C, Macias W, Rajadhyaksha A, Carlson RR, Barczak A, Kang S et al. J Neurosci 2000; 20: 4011–4020. 4 Melone M, Vitellaro-Zuccarello L, Vallejo-Illarramendi A, Perez-Samartin A, Matute C, Cozzi A et al. Mol Psychiatry 2001; 6: 380–386. 5 Lewis DA, Gonzalez-Burgos G. Nature Med 2006; 12: 1016–1022. 6 Wojcik SM, Katsurabayashi S, Guillemin I, Friauf E, Rosenmund C, Brose N et al. Neuron 2006; 50: 575–587. 7 De Gois S, Schafer MK, Defamie N, Chen C, Ricci A, Weihe E et al. J Neurosci 2005; 25: 7121–7133. 8 Bragina L, Melone M, Fattorini G, Torres-Ramos M, VallejoIllarramendi A, Matute C et al. J Neurochem 2006; 99: 131–141. 9 Chaudry FA, Reimer RJ, Bellocchio EE, Danbolt NC, Osen KK, Edwards RH et al. J Neurosci 1998; 18: 9733–9750. 10 Minelli A, Alonso-Nanclares L, Edwards RH, DeFelipe J, Conti F. Neuroscience 2003; 117: 337–346.