James E. Koch

Central opioid receptor subtype antagonists differentially reduce intake of saccharin and maltose dextrin solutions in rats

Opioid modulation of ingestion includes general opioid antagonism of deprivation-induced water intake and intake of sucrose and saccharin solutions. Previous studies using selective subtype antagonists indicated that opioid effects upon deprivation-induced water intake occurred through the mu2 receptor and that opioid effects upon sucrose intake occurred through kappa and mu2 receptors. The present study compared the effects of intracerebroventricular administration of opioid receptor subtype antagonists upon intakes of a saccharin solution and a maltose dextrin (MD) solution to determine which receptor subtypes were involved in modulation of ingestion of different preferred tastants. Significant reductions in saccharin intake (1 h) occurred following naltrexone (20-50 micrograms: 66%) and naltrindole (delta, 20 micrograms: 75%), whereas [D-Ala2, Leu5, Cys6]-enkephalin (DALCE, delta 1, 40 micrograms: 45%) had transient (5 min) effects. Neither beta-funaltrexamine (B-FNA, mu), naloxonazine (mu1), nor nor-binaltorphamine (Nor-BNI, kappa) significantly altered saccharin intake. Significant reductions in MD intake (1 h) occurred following naltrexone (5-50 micrograms: 69%) and B-FNA (1-20 micrograms: 38%). MD intake was not reduced by naltrindole, DALCE, naloxonazine and Nor-BNI. Peak antagonist effects were delayed (20-25 min) to reflect interference with the maintenance, rather than the initiation of saccharin or MD intake. Comparisons of opioid antagonist effects across intake situations revealed that naltrexone had consistently low ID40 values for saccharin (29 nmol), MD (25 nmol), sucrose (6 nmol) and deprivation (38 nmol) intake. Despite its significant effects relative to naloxonazine, B-FNA had significantly higher ID40 values for saccharin (800 nmol), MD (763 nmol) and sucrose (508 nmol) relative to deprivation (99 nmol) intake, suggesting that mu2 receptors may be mediating maintenance of intake rather than taste effects. Nor-BNI had low ID40 values for intake of sucrose (4 nmol), but not for saccharin (168 nmol), MD (153 nmol) and deprivation (176 nmol), suggesting that kappa receptors may mediate ingestion of sweet-tasting stimuli. That delta (naltrindole: ID40 = 60 nmol), but not delta 1 (DALCE: ID40 = 288 nmol) antagonists consistently reduce saccharin intake suggests a role for the delta 2 receptor subtype in the modulation of hedonic orosensory signals.

Selective alterations in macronutrient intake of food-deprived or glucoprivic rats by centrally-administered opioid receptor subtype antagonists in rats

Two hypotheses have attempted to account for the abilities of opioid agonists and antagonists to respectively stimulate and inhibit food intake in rats. The first suggests that the opioid system selectively modulates fat intake, while the second suggest that the opioid system selectively alters intake of that macronutrient which the animal prefers. The present study evaluated these two hypotheses by examining total intake and individual macronutrient intake in either food-deprived (24 h) rats or rats made glucoprivic with 2-deoxy-D-glucose (2DG, 200 mg/kg, i.p.) following either vehicle treatment, systemic administration of naltrexone or intracerebroventricular administration of either naltrexone, the mu opioid antagonist, beta-funaltrexamine (B-FNA), the mu1 opioid antagonist, naloxonazine, the kappa opioid antagonist, nor-binaltorphamine (Nor-BNI), the delta opioid antagonist, naltrindole or the delta1 opioid antagonist, DALCE. Systemic administration of naltrexone (0.5-5 mg/kg significantly reduced carbohydrate, fat and total intake in deprived rats, and carbohydrate, fat, protein and total intake in glucoprivic rats. Central administration of naltrexone (5-50 micrograms) significantly reduced fat and total intake in both deprived and glucoprivic rats. B-FNA (5-20 micrograms) significantly reduced carbohydrate, fat and total intake in both deprived and glucoprivic rats Naloxonazine (10-100 micrograms) significantly reduced carbohydrate, fat and total intake in deprived rats, but failed to alter 2DC intake. Nor-BNI (5-20 micrograms) significantly reduced fat and total intake in glucoprivic rats, but failed to alter deprivation intake. Neither naltrindole (20 micrograms) nor DALCE (40 micrograms altered intake in deprived or glucoprivic rats. Carbohydrate or fat preference in deprived rats significantly increased the amount of explained variance in the inhibitory actions of central naltrexone, B-FNA and naloxonazine upon deprivation-induced intake. Carbohydrate or fat preference in glucoprivic rats significantly increased the amount of explained variance in the inhibitory action of systemic and central naltrexone, B-FNA, naloxonazine and Nor-BN upon 2-DG hyperphagia. These data are discussed in terms of the contentions that opioids either selectively alter fat intake pe se or selectively alter the preferred macronutrient.

Alterations in deprivation, glucoprivic and sucrose intake following general, mu and kappa opioid antagonists in the hypothalamic paraventricular nucleus of rats

While opioid agonists administered into the hypothalamic paraventricular nucleus increase food intake in rats, naloxone reduces deprivation-induced intake. Ventricular administration of either mu (beta-funaltrexamine) or kappa (nor-binaltorphamine) opioid antagonists reduces spontaneous, deprivation, glucoprivic and palatable intake. The present study assessed whether microinjections of either general, mu or kappa opioid antagonists into the paraventricular nucleus altered either deprivation (24 h) intake, 2-deoxy-D-glucose hyperphagia or sucrose intake in rats. Deprivation intake was significantly reduced by nor-binaltorphamine (5 micrograms, 68 nmol, 30-33%), beta-funaltrexamine (5 micrograms, 100 nmol, 26-29%) or naltrexone (10 micrograms, 260 nmol, 26%) in the paraventricular nucleus. 2-Deoxy-D-glucose hyperphagia was significantly reduced only after 2 h by naltrexone (10 micrograms, 260 nmol, 69%), norbinaltorphamine (20 micrograms, 272 nmol, 69%) or beta-funaltrexamine (20 micrograms, 400 nmol, 83%) in the paraventricular nucleus. Sucrose intake was significantly reduced by nor-binaltorphamine (5 micrograms, 68 nmol, 27-36%), naltrexone (5-10 micrograms, 130-260 nmol, 18-31%) and beta-funaltrexamine (5 micrograms, 100 nmol, 20%) in the paraventricular nucleus. These data indicate that general, mu and kappa opioid antagonists administered into the hypothalamic paraventricular nucleus produce similar patterns of effects upon different forms of food intake as did ventricular administration, implicating this nucleus as part of the circuitry underlying opioid mediation of ingestion.

Involvement of mu1 and mu2 opioid receptor subtypes in tail-pinch feeding in rats

Tail-pinch feeding (TPF) in rats is decreased following general (naltrexone, NTX) and mu (Cys2-Tyr3-Orn5-Pen7-amide, CTOP) opioid antagonists, but not following kappa (nor-binaltorphamine. Nor-BNI) or delta (naltrindole, NTI) opioid antagonists. Because multiple mu (mu1 and mu2) and delta (delta 1 and delta 2) opioid receptor subtypes have been characterized, the present study evaluated whether TPF was differentially altered following ICV administration of general (NTX), mu (beta-funaltrexamine, B-FNA), mu1 (naloxonazine, NAZ), kappa (Nor-BNI), delta 1 ([D-Ala2, Leu5, Cys6]-enkephalin, DALCE) and delta 2 (NTI) opioid antagonists. Like the reversible mu antagonist CTOP, the irreversible mu antagonist B-FNA significantly and dose-dependently (1-20 micrograms) reduced TPF by up to 28%. In contrast, whereas NAZ (50 micrograms) reduced TPF by 32%, this effect was highly variable and failed to achieve significance. Neither NTX (5-10 mg/kg, SC), Nor-BNI (20 micrograms), DALCE (40 micrograms) nor NTI (20 micrograms) significantly altered TPF, suggesting that kappa, delta 1 and delta 2 opioid receptor subtypes were not involved. Because no antagonist altered the duration of food contact during tail pinch, it appears that the opioid effect modulates ingestive rather than activational mechanisms. The reliable inhibition of TPF by B-FNA (mu1 and mu2), together with the variable effect of naloxonazine (mu1), appears to implicate both mu binding sites in this response.

Differential modulation of angiotensin II and hypertonic saline-induced drinking by opioid receptor subtype antagonists in rats

Opioid modulation of ingestion includes general opioid antagonism of different forms of water intake, mu 2 receptor modulation of deprivation-induced water intake and delta 2 receptor modulation of saccharin intake. Water intake is stimulated by both central administration of angiotensin II (ANG II) and peripheral administration of a hypertonic saline solution; both responses are reduced by general opioid antagonists. The present study examined whether specific opioid receptor subtype antagonists would selectively alter each form of water intake in rats. Whereas systemic naltrexone (0.1-2.5 mg/kg, s.c.) reduced water intake induced by either peripheral ANGII (500 micrograms/kg, s.c.) or hypertonic saline (3 ml/kg, 10%), intracerebroventricular (i.c.v.) naltrexone (1-50 micrograms) only inhibited central ANGII (20 ng)-induced hyperdipsia. Both forms of drinking were significantly and dose-dependently inhibited by the selective kappa antagonist, nor-binaltorphamine (Nor-BNI, 1-20 micrograms). Whereas both forms of drinking were transiently reduced by the mu-selective antagonist, beta-funaltrexamine (beta-FNA, 1-20 micrograms), the mu 1 antagonist, naloxonazine (40 micrograms) stimulated drinking following hypertonic saline. The delta 1 antagonist, [D-Ala2, Leu5, Cys6]-enkephalin (DALCE, 1-40 micrograms) significantly reduced drinking following ANGII, but not following hypertonic saline; the delta antagonist, naltrindole failed to exert significant effects. These data indicate that whereas kappa opioid binding sites modulate hyperdipsia following hypertonic saline, mu 2, delta 1, and kappa opioid binding sites modulate hyperdipsia following ANGII. The mu 1 opioid binding site may normally act to inhibit drinking following saline.

Analysis of central opioid receptor subtype antagonism of hypotonic and hypertonic saline intake in water-deprived rats

Intake of either hypotonic or hypertonic saline solutions is modulated in part by the endogenous opioid system. Morphine and selective mu and delta opioid agonists increase saline intake, while general opioid antagonists reduce saline intake in rats. The present study evaluated whether intracerebroventricular administration of general (naltrexone) and selective mu (beta-funaltrexamine, 5-20 micrograms), mu, (naloxonazine, 50 micrograms), kappa (nor-binaltorphamine, 5-20 micrograms), delta (naltrindole, 20 micrograms), or delta 1 (DALCE, 40 micrograms) opioid receptor subtype antagonists altered water intake and either hypotonic (0.6%) or hypertonic (1.7%) saline intake in water-deprived (24 h) rats over a 3-h time course in a two-bottle choice test. Whereas peripheral naltrexone (0.5-2.5 mg/kg) significantly reduced water intake and hypertonic saline intake, central naltrexone (1-50 micrograms) significantly reduced water intake and hypotonic saline intake. Water intake was significantly reduced following mu and kappa receptor antagonism, but not following mu 1, delta, or delta 1 receptor antagonism. In contrast, neither hypotonic nor hypertonic saline intake was significantly altered by any selective antagonist. These data are discussed in terms of opioid receptor subtype control over saline intake relative to the animals hydrational state and the roles of palatability and/or salt appetite.

Naloxone benzoylhydrazone, a κ3 opioid agonist, stimulates food intake in rats

Abstract Naloxone benzoylhydrazone (NalBzoH) is a selective, short-acting agonist at theκ 3 opioid receptor and a slowly dissociating potent antagonist at the mu opioid receptor. Given the important role of κ receptors in the opioid control of food intake, the present study examined the central and peripheral effects of NalBzoH upon food intake. Central administration of NalBzoH (1–20 μg, i.c.v.) significantly increased food intake for up to 12 h, but failed to alter intake or body weight after 24 or 48 . The 12 h duration of NalBzoH-mediated effects may be due to either persistentκ 3 receptor occupancy, and/or activation of an ingestive system which maintains its activity. Peripheral administration of NalBzoH (20 mg/kg, s.c.) significantly increased food intake for up to 1 h. To distinguishκ 1 (U50, 488H) andκ 3 (NalBzoH) hyperphagic effects, these agonist effects were compared following pretreatment with either naltrexone or theκ 1 antagonist, nor-binaltorphamine (Nor-BNI). Whereas naltrexone significantly reduced both U50, 448H and NalBzoH hyperphagia, Nor-BNI blocked U50, 448H, but not NalBzoH hyperphagia. These data indicate a distinct role for theκ 3 receptor in ingestive behavior separable from thatofκ 1 effects.

Naltrexone, serotonin receptor subtype antagonists, and carbohydrate intake in rats.

Functional interactions between serotonergic (5-HT) and opioid drugs have been observed with 5-HT3 receptor antagonism enhancing the inhibitory actions of naloxone and naltrexone in both food-deprived and glucoprivic rats; 5-HT2A/C receptor antagonism enhanced naltrexones inhibition of insulin hyperphagia. The present study examined whether pretreatment with either general 5-HT (methysergide: 0.5-5 mg/kg), 5-HT2A/C (ritanserin: 0.25-2.5 mg/kg), or 5-HT3 (ICS 205930: 0.5-5 mg/kg) antagonists altered the pattern and magnitude of ad lib intake of simple (sucrose: 10%) or more complex (maltose dextrin: MD, 10%) carbohydrate solutions, or naltrexones (0.25-2.5 mg/kg) inhibition of these forms of intake. Methysergide significantly increased the pattern and magnitude of sucrose intake at low (0.5-2.5 mg/kg) doses, and transiently delayed the pattern of MD intake at high (5 mg/kg) doses. Ritanserin significantly accelerated the pattern, but not the magnitude of sucrose intake at low (0.25-1.25 mg/kg) doses without affecting MD intake. ICS 205930 reduced the magnitude of sucrose intake at the highest (5 mg/kg) dose, and transiently reduced MD intake. Naltrexone dose dependently altered the pattern and magnitude of both sucrose and MD intake. Coadministration of ritanserin and naltrexone either eliminated or delayed the pattern of opioid antagonist inhibition of both sucrose and MD intake. Methysergide and ICS 205930 pretreatment produced minor changes in the pattern of naltrexone-induced inhibition. These data indicate that 5-HT receptor differentially modulate the pattern of carbohydrate intake, and indicate differential ingestive interactions between 5-HT and opioid antagonists under challenge and palatable conditions.

Naltrexone, dopamine receptor agonists and antagonists, and food intake in rats: 1. Food deprivation

Different forms of food intake are reduced by both agonists and antagonists of dopamine D1 and D2 receptors as well as general opioid antagonists. The present study evaluated whether deprivation (24 h)-induced food intake was altered following systemic administration of either the D1 agonist, SKF-38393, the D1 antagonist, SCH-23390, the D2 agonist, quinpirole, or the D2 antagonist, haloperidol, alone or in combination with the general opioid antagonist, naltrexone. Both SKF-38393 (5-10 mg/kg) and SCH-23390 (100-200 micrograms/kg) significantly and dose dependently reduced deprivation-induced intake. Whereas quinpirole (0.5-1 mg/kg) failed to alter deprivation-induced intake, haloperidol increased deprivation-induced intake at low (50 micrograms) doses and decreased intake at higher (100-500 micrograms/kg) doses. Naltrexone (2.5-10 mg/kg) significantly inhibited deprivation-induced intake. When naltrexone was paired with behaviorally ineffective doses of either SCH-23390 (2.5-100 micrograms/kg), quinpirole (0.01-1 mg/kg), or haloperidol (50 micrograms/kg), the degree of reduction of deprivation-induced intake was significantly greater than that produced by naltrexone alone. Pairing naltrexone with SKF-38393 produced reductions of deprivation-induced intake comparable to that of naltrexone alone.

Naltrexone, serotonin receptor subtype antagonists, and glucoprivic intake: 1.2-deoxy-D-glucose

Inhibition of deprivation-induced intake by naloxone was significantly enhanced by the 5-hydroxytryptamine3 (5-HT3) antagonist ICS-205,930. Interactions between naloxone and either the general 5-HT antagonist methysergide or the 5-HT2 antagonist ritanserin or ketanserin produced smaller effects. The present study evaluated whether 2-deoxy-D-glucose (2DG, 400 mg/kg) hyperphagia was affected by methysergide (0.5-5 mg/kg), ritanserin (0.25-2.5 mg/kg), or ICS-205,930 (0.5-5 mg/kg) alone or in combination with naltrexone (0.25 and 2.5 mg/kg). Only ICS-205,930 stimulated spontaneous intake for up to 4 h in the light cycle. Only ritanserin (1.25 mg/kg) transiently reduced 2DG hyperphagia. The dose-dependent decreases in 2DG hyperphagia by naltrexone were significantly enhanced by the dose range of ICS-205,930. The inhibition of 2DG hyperphagia by the low naltrexone dose was enhanced by methysergide (5 mg/kg) and ritanserin (1.25 mg/kg). These data suggest that the 5-HT3 receptor primarily interacts with opioid systems to modulate 2DG hyperphagia and that one possible locus of interaction is in the caudal brainstem.