GUTIÉ RREZ, ARANTZA, GONZALO SARACÍBAR, LUIS CASIS, ENRIQUE ECHEVARRÍA, VÍCTOR MAN-UEL RODRÍGUEZ, MARIA TERESA MACARULLA, LUIS C. ABECIA, AND MARÍA P. PORTILLO. Effects of fluoxetine administration on neuropeptide Y and orexins in obese Zucker rat hypothalamus. Obes Res. 2002;10: 532-540. Objective: The aim of this work was to study the potential involvement of neuropeptide Y (NPY) and orexins in the anorexigenic mechanism of fluoxetine in obese Zucker rats, assessing the effects of chronic fluoxetine treatment on NPY and orexin immunostaining in several hypothalamic regions. Research Methods and Procedures: Male obese Zucker (fa/fa) rats were administered fluoxetine (10 mg/kg intraperitoneally) daily for 2 weeks. The control group was administered 0.9% NaCl solution. Carcass composition was assessed using the official methods of the Association of Official Analytical Chemists. To test the potential thermogenic effect of fluoxetine administration, total body oxygen consumption was measured daily for 60 minutes before fluoxetine or saline injection and for 30 minutes after drug or saline injection. Hypothalamic arcuate and paraventricular nuclei, and the lateral hypothalamic area were immunostained for NPY, orexin A, and orexin B. Commercial kits were used for serum determinations. Results: Chronic fluoxetine administration in obese Zucker rats generated a reduction in body weight gain, food intake, adipocyte size, fat mass, and body protein. A decrease in NPY immunostaining in the paraventricular nucleus, without changes in the arcuate, was observed. However, no changes were observed in the number of neural cells immunostained for orexin A or orexin B in the lateral hypothalamic area. Discussion: Due to the hyperphagic effect of NPY in the paraventricular nucleus, these results suggest that NPY, but not orexins, could be involved in the anorexigenic effect of fluoxetine in obese Zucker rats.

Therefore, chronic nefazodone induces antinociceptive and antidepressant-like effects in rats and increases mu-opioid receptor expression in brain areas related to pain and affective state. These results suggest that antidepressants could be effective on somatic and affective dimensions of pain and this action could be related to its influence on the opioid system.

Toluene is an ototoxic organic solvent widely used in industry and could be a cause of sleep apnea. Acute toluene administration in rats induces an increase in the number of neural cells immunostained for mu-opioid receptors in several brainstem nuclei, such as the inferior colliculus, dorsal and lateral periaqueductal gray and dorsal raphe, without changes in the superior colliculus and the interpeduncular and lateral reticular nuclei. These data suggest that mu-opioid receptors could be involved in toluene-induced neurotoxic effects on the physiological regulation of breathing during sleep, and auditive function. Key words: Toluene, Mu-opioid receptors, Rat brain Materials and MethodsThe experimental animals were carefully handled and all experiments were carried out in accordance with the law, avoiding animal suffering. Toluene with purity of 99% (Analytical reagent grade, Quimon Chem. Co., Spain) was used. Toluene was diluted with olive oil at a concentration of 1 ml/ml and administered intraperitoneally to the experimental group (n=6) at a dose of 1.3 ml/kg/day for 5 consecutive days. The selected dose was 1/2 of the LD 50 per day. This dose and this kind of administration have been employed to carry out studies on the adverse effects of organic solvents 15) . The LD 50 in rats has been found in our laboratory to be 2.61 ± 0.41 ml/kg/day, calculated by the Bliss method. The control group (n=6) was given 0.9% NaCl solution in the same volume and duration as the experimental group. Two hours after the last treatment, the animals were anaesthetized with Equithensin (2 ml/kg), an alcoholic solution of nembutal and chloral hydrate (Sigma-Aldrich Química S. A.), intraperitoneally and perfused transcardially under deep anaesthesia with saline plus 50 mM phosphate buffer, pH 7.4, followed by 4% paraformaldehyde (Sigma-Aldrich Química S. A.). The brains were removed, cut into smaller pieces and then immersed in the same fixative medium overnight. They were stored for 2 days in 0.1 M phosphate buffer containing

Imipramine (CAS 113-52-0) is being utilized widely for the treatment of major depression. In recent years, there has been evidence of the involvement of the endogenous opioid system in major depression and its treatment. There is some evidence indicating that opioid receptors could be involved in the antidepressant mechanism of action. Regarding this topic, mood-related behavior of endogenous enkephalins seems to be mediated by delta-opioid receptors. In this work, the effects of subacute (5 day) and chronic (15 day) treatments of imipramine on the density and the affinity of the delta-receptors in the striatum and in the parietal and frontal cortices of the rat brain are described. Studied parameters (Bmax and Kd) were calculated by a saturation binding assay with the delta-opioid agonists [3H]-DPDPE (tyrosyl-2,6-3H(N)-(2-D-penicillamine-5-D-penicillamine)-enkephalin) as specific ligand and DSLET ([D-serine2]-D-leucine-enkephalin-threonine) as non-radioactive competing ligand. It was found that 15 days treatment significantly decreased the delta-opioid receptor density,without changing the affinity, in the frontal cortex of the rat brain. That decrease was confirmed by delta-opioid receptor immunostaining. These results suggest that delta-opioid receptors could play a role in the chronic action mechanism of imipramine.

Nefazodone is an antidepressant drug that inhibits serotonin and noradrenaline reuptake. The aim of the present work was to study the effects of nefazodone on food intake, body weight, adiposity and hypothalamic NPY immunostaining in rats. For this purpose, male Sprague-Dawley rats (3-month-old) were administered with nefazodone (20 mg/kg; i.p) daily for two weeks. The control group was given 0.9% NaCl solution. Hypothalamic arcuate, paraventricular, periventricular, supraoptic and suprachiasmatic nuclei and the lateral hypothalamic area were immunostained for NPY. Chronic nefazodone administration in rats did not modify food intake, body weight and adipose depot size (subcutaneous, perirenal and epididymal white adipose tissues, and interscapular brown adipose tissue). However, a significant decrease in paraventricular NPY immunostaining was found in the nefazodone group compared with the control group. No changes in other hypothalamic regions such as arcuate, periventricular, supraoptic and suprachiasmatic nuclei, and lateral and medial preoptic areas were observed. Because nefazodone is an alpha1-adrenoceptor antagonist, it can be proposed that the expected decrease in food intake after nefazodone administration, due to its effects on NPY arcuate-paraventricular projection, could be masked by the opposite orexigenic effect of alpha1-adrenoceptor blockade.