Gaylen L. Edwards

Disulfiram attenuates drug-primed reinstatement of cocaine seeking via inhibition of dopamine β-hydroxylase.

The antialcoholism medication disulfiram (Antabuse) inhibits aldehyde dehydrogenase (ALDH), which results in the accumulation of acetaldehyde upon ethanol ingestion and produces the aversive ‘Antabuse reaction’ that deters alcohol consumption. Disulfiram has also been shown to deter cocaine use, even in the absence of an interaction with alcohol, indicating the existence of an ALDH-independent therapeutic mechanism. We hypothesized that disulfirams inhibition of dopamine β-hydroxylase (DBH), the catecholamine biosynthetic enzyme that converts dopamine (DA) to norepinephrine (NE) in noradrenergic neurons, underlies the drugs ability to treat cocaine dependence. We tested the effects of disulfiram on cocaine and food self-administration behavior and drug-primed reinstatement of cocaine seeking in rats. We then compared the effects of disulfiram with those of the selective DBH inhibitor, nepicastat. Disulfiram, at a dose (100 mg/kg, i.p.) that reduced brain NE by ∼40%, did not alter the response for food or cocaine on a fixed ratio 1 schedule, whereas it completely blocked cocaine-primed (10 mg/kg, i.p.) reinstatement of drug seeking following extinction. A lower dose of disulfiram (10 mg/kg) that did not reduce NE had no effect on cocaine-primed reinstatement. Nepicastat recapitulated the behavioral effects of disulfiram (100 mg/kg) at a dose (50 mg/kg, i.p.) that produced a similar reduction in brain NE. Food-primed reinstatement of food seeking was not impaired by DBH inhibition. Our results suggest that disulfirams efficacy in the treatment of cocaine addiction is associated with the inhibition of DBH and interference with the ability of environmental stimuli to trigger relapse.

Lesion of the lateral parabrachial nucleus attenuates the anorectic effect of peripheral amylin and CCK

Amylin and CCK activate the area postrema (AP)/nucleus of the solitary tract (NTS) - lateral parabrachial nucleus (LPBN) - central amygdala (CeA) pathway. However, except for the brainstem structures the role of these nuclei for the anorectic effect of these peptides is not yet well characterized. The current study investigated the role of the LPBN in mediating the inhibitory effect of peripheral amylin and CCK on feeding behavior. Rats with electrolytic lesions in the LPBN (LPBN-X) were used in behavioral as well as in immunohistological c-Fos studies. LPBN-X significantly reduced the anorectic effect of amylin (5 microg/kg, i.p.). The effect of a higher amylin dose (10 microg/kg, i.p.) was only slightly attenuated in the LPBN-X rats. In agreement with previous studies, LPBN lesions also reduced the inhibitory effect of CCK on food intake. In the immunohistological experiments, amylin and CCK induced c-Fos expression in the AP, NTS, LPBN and CeA in the SHAM rats. Both the amylin- and CCK-induced activation of the CeA was completely abolished in the animals with a LPBN lesion. These results clearly suggest that the signal transduction pathway between the AP/NTS and CeA has been disrupted by the LPBN ablation. We conclude that the LPBN is a crucial brain site mediating the anorectic effect of amylin and CCK. Furthermore, an intact LPBN seems to be essential for the c-Fos response in the CeA induced by these peptides.

Cannabinoid agonist, CP 55,940, facilitates intake of palatable foods when injected into the hindbrain.

Cannabinoids have been shown to influence food intake, and until recently, the neural pathways mediating these effects have remained obscure. It has been previously shown that intracerebroventricular injection of delta-9-tetrahydrocannabinol (Delta(9)-THC) causes increased consumption of palatable foods in rats, and we postulated the involvement of the hindbrain in this cannabinoid-induced food intake. Cannulated rats (both female and male groups) trained to consume sweetened condensed milk received either lateral or fourth ventricle injections of CP 55,940 and were presented with sweetened condensed milk 15 min after injection. Rats were injected over a range of doses between 100 pg and 10 microg per rat. Milk intake was recorded for a total of 3 h. Lateral ventricle injection of CP 55,940 increased milk intake at doses in the microgram range. However, CP 55,940 was effective in increasing food intake at nanogram doses when injected into the fourth ventricle. Finally, male rats appeared to be more sensitive to CP 55,940 than female rats inasmuch as milk consumption was increased at the 1 ng dose in male rats, whereas only the 10 ng dose was effective in females. These results indicate that CP 55,940 may act in the hindbrain to influence feeding behavior in rats.

Neuropeptide Y antibody attenuates 2-deoxy-D-glucose induced feeding in rats

2-deoxy-D-glucose (2-DG) has been shown to induce increased feeding responses in animals. Recent studies suggest the possible involvement of neuropeptide Y (NPY) in 2-DG-induced feeding. The present study examined the effect of immunoneutralization of endogenous NPY on 2-DG-induced feeding. NPY antibody injected into the paraventricular nucleus of the rats significantly attenuated 2-DG-induced feeding, suggesting that hypothalamic NPY may mediate, at least partly, the effect of 2-DG on food intake.

Intracerebroventricular Injection of Fructose Stimulates Feeding in Rats

Abstract 2-Deoxy-d-glucose (2DG) inhibits glycolysis and stimulates food intake. Previous work suggests that fructose may attenuate the hyperglycemic and hypothermic effects of 2DG. The current study examined the effect of intracerebroventricular fructose on 2DG-induced feeding. We found that concentrated fructose injected into the cerebroventricles enhanced food intake both in the presence and absence of 2DG. On the other hand, similar concentrations of glucose suppressed 2DG-induced food intake. These data suggest differences in metabolism of glucose and fructose and may provide insight into the metabolic steps monitored by brain glucoreceptors to control food intake.

Effects of neurotoxic destruction of descending noradrenergic pathways on cannabinoid antinociception in models of acute and tonic nociception

The effects of neurotoxic destruction of catecholaminergic projections to the spinal cord on cannabinoid antinociception were examined in models of acute and tonic nociception. High performance liquid chromatography was used to quantify monoamine levels in sham-operated and lesioned rats. Intrathecal administration of the catecholamine neurotoxin 6-hydroxydopamine (6-OHDA) induced a selective depletion of norepinephrine (by approximately 85% of control) in rat lumbar spinal cord without altering levels of dopamine or serotonin. By contrast, brain levels of monoamines did not differ in sham-operated and lesioned rats. Pain behavior was similar in sham-operated and lesioned rats receiving vehicle in models of both acute and tonic nociception. The cannabinoid agonist WIN55,212-2 (5 or 10 mg/kg, i.p.) produced antinociception in the tail-flick test in sham-operated rats. The antinociceptive effect of WIN55,212-2 was attenuated relative to control conditions in rats depleted of spinal norepinephrine. WIN55,212-2 suppressed tonic pain behavior in the formalin test in sham-operated rats during phase 2 (15-60 min post formalin) of nociceptive responding. By contrast, in lesioned rats, WIN55,212-2 suppressed pain behavior during phase 1 (0-9.9 min) and phase 2A (10-39.9 min), but not during phase 2B (40-60 min). The cannabinoid agonist suppressed formalin-evoked Fos protein expression, a marker of neuronal activity, in the lumbar dorsal horn of sham-operated rats, but no suppression was observed in lesioned rats. The number of formalin-evoked Fos-like immunoreactive (FLI) cells was greater in lamina I and II of lesioned rats relative to sham-operated rats. These data indicate that the suppressive effect of the cannabinoid on formalin-evoked Fos protein expression in the superficial dorsal horn was attenuated following destruction of descending noradrenergic pathways. Our data are consistent with the hypothesis that cannabinoids produce antinociception, in part, by modulating descending noradrenergic systems and support a differential involvement of noradrenergic projections to the spinal cord in cannabinoid modulation of acute versus tonic nociception.

Area postrema lesions elevate NPY levels and decrease anxiety-related behavior in rats.

Intracerebroventricular administration of neuropeptide Y (NPY) has been shown to reduce anxiety-like effects in rodents and also profoundly alter feeding. The area postrema-lesioned (APX) rat model of food motivated behavior overconsumes palatable foods and expresses significantly higher levels of NPY in the hypothalamus than sham-lesioned control rats. For this reason, we examined APX rats in the open field test, which is a standard measure of anxiety- or fear-related behavior and also investigated NPY mRNA levels in the hippocampus, amygdala and hypothalamus. We found that APX rats display reduced anxiety-like behavior in the open field test as indicated by spending increased time in the center of the field as opposed to the perimeter of the field. NPY mRNA levels were also found to be significantly elevated in the amygdala, hippocampus and arcuate nucleus of the hypothalamus of APX rats when compared to sham-lesioned rats. These results support the action of limbic NPY to reduce anxiety-like behavior in a rodent model that appears to express higher than normal NPY levels.

Galanin mediates features of neural and behavioral stress resilience afforded by exercise.

Exercise promotes resilience to stress and increases galanin in the locus coeruleus (LC), but the question of whether changes in galanin signaling mediate the stress-buffering effects of exercise has never been addressed. To test the contributions of galanin to stress resilience, male Sprague Dawley rats received intracerebroventricular (ICV) cannulation for drug delivery and frontocortical cannulation for microdialysis, and were housed with or without a running wheel for 21d. Rats were acutely injected with vehicle or the galanin receptor antagonist M40 and exposed to a single session of either footshock or no stress. Other groups received galanin, the galanin receptor antagonist M40, or vehicle chronically for 21d prior to the stress session. Microdialysis sampling occurred during stress exposure and anxiety-related behavior was measured on the following day in the elevated plus maze. Dendritic spines were visualized by Golgi impregnation in medial prefrontal cortex (mPFC) pyramidal neurons and quantified. Exercise increased galanin levels in the LC. Under non-stressed conditions, anxiety-related behavior and dopamine levels were comparable between exercised and sedentary rats. In contrast, exposure to stress reduced open arm exploration in sedentary rats but not in exercise rats or those treated chronically with ICV galanin, indicating improved resilience. Both exercise and chronic, ICV galanin prevented the increased dopamine overflow and loss of dendritic spines observed after stress in sedentary rats. Chronic, but not acute M40 administration blocked the resilience-promoting effects of exercise. The results indicate that increased galanin levels promote features of resilience at both behavioral and neural levels.

The area postrema is involved in paraquat-induced conditioned aversion behavior and neuroendocrine activation of the hypothalamic-pituitary-adrenal axis

Paraquat is a herbicide capable of eliciting conditioned taste aversion (CTA), a behavioral response characteristic of toxicosis. The area postrema (AP) is a hindbrain circumventricular organ previously shown to be important in mediating signs of paraquat-induced toxicity, namely CTA and weight loss. The relationship between neural substrates for paraquat-induced CTA and activation of the hypothalamic-pituitary-adrenal (HPA) axis was investigated in Sprague-Dawley rats with lesions centered on the AP (APX) and sham-operated (SHM) rats administered paraquat (25 mumol/kg) or saline (1 ml/kg). Injection of paraquat at a dose sufficient to condition taste aversion, but produce no other signs of overt toxicity, significantly increased plasma corticosterone concentrations in SHM rats up to 4 h after administration. Paraquat-induced activation of the HPA axis was significantly attenuated in AP-lesioned rats as compared to sham-operated controls. These findings suggest the area postrema is a common neural substrate for the behavioral and neuroendocrine responses to paraquat.

Glucagon-like peptide-1 (7-36) amide administered into the third cerebroventricle inhibits water intake in rats.

Abstract Intracerebroventricular (icv) injection of glucagon-like peptide-1 (7–36) amide (GLP-1) has been shown to reduce food intake in rats. In these studies, we confirmed that injection of 10 μg of GLP-1 into the third cerebroventricle suppressed food intake. Moreover, we observed a reduction in water intake associated with the decreased food intake. We further examined whether GLP-1 injected icv in rats has a specific inhibitory effect on water intake. It was found that GLP-1 reduced water deprivation-induced drinking. Furthermore, the same dose of GLP-1 (10 μg) was sufficient to condition taste aversion. Finally, when 2 μg of GLP-1 were injected into the third ventricle, it only suppressed water deprivation-induced water intake and failed to influence spontaneous food and water intakes or induce conditioned taste aversion. These observations indicate that GLP-1 is a potent inhibitor of water intake in the rat and may play a role in the control of fluid homeostasis.