Attenuation of the discriminative stimulus effects of ethanol by the benzodiazepine partial inverse agonist Ro 15-4513

The aim of the present study was to investigate whether ethanol training affects the ability of Ro 15-4513 to block the discriminative stimulus effects of ethanol dose differentially. Three different groups of rats were trained to discriminate 1.0 g/kg ethanol (n = 8), 1.5 g/kg ethanol (n = 7) or 2.0 g/kg ethanol (n = 8) from water in a two-lever, food-reinforced procedure. Ethanol and water were administered by gavage 20 min before the onset of the session. When the discrimination performance was stable, rats were pretreated with Ro 15-4513 (1-17 mg/kg; i.p.) 5 min before the administration of ethanol. Ro 15-4513 attenuated the discriminative stimulus effects of 1.0 and 1.5 g/kg ethanol but not 2.0 g/kg ethanol in each of the ethanol training groups. Overall, blockade of the discriminative stimulus effects of 1.0 and 1.5 g/kg ethanol by 5.6 mg/kg Ro 15-4513 occurred without significantly altering response rates or blood ethanol concentrations. A decrease in blood ethanol concentration was, however, found with 17 mg/kg Ro 15-4513 in combination with 2.0 g/kg ethanol. These results suggest that the benzodiazepine partial inverse agonist, Ro 15-4513, can attenuate the discriminative stimulus effects associated with low to moderate doses of ethanol (1.0-1.5 g/kg).     

Effects of benzodiazepine receptor ligands on the ingestion of sucrose, intralipid, and maltodextrin: An investigation using a microstructural analysis of licking behavior in a brief contact test.

Microstructural analysis of licking behavior in the rat was conducted (a) to describe in detail the characteristics of benzodiazopine-induced changes in ingestion and (b) to determine if the changes are consistent with an alteration in palatability. The effects of the benzodiazepine receptor (BZR) agonist midazolam (0.3–3 mg/kg), and the partial inverse agonist Ro 15-4513 (0.3–3 mg/kg), on licking for several concentrations of sucrose, Intralipid, and maltodextrin in a brief contact test were investigated. Midazolam increased the total number of licks for all 3 fluids; conversely, Ro 15-4513 decreased the total number of licks. Midazolam increased mean bout duration for sucrose and maltodextrin drinking and there was a trend toward a similar effect with Intralipid drinking. Ro 15-4513 reduced mean bout duration for all 3 test fluids. These data are discussed in terms of bidirectional changes in fluid palatability by drug actions at BZRs. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Inactivation of the median raphe nucleus increases intake of sucrose solutions: A microstructural analysis.

Previous studies have shown that microinjections of the GABA-A agonist muscimol into the median raphe nucleus (MR) result in large increases in the intake of solid foods. In the current study, we used microstructural techniques to characterize the effects of intra-MR muscimol injections on the consumption of either a 0.05 M or a 0.29 M sucrose solution. After injections of either saline or muscimol, animals consumed more of the 0.29 M than the 0.05 M solution, an effect which resulted primarily from increases in the initial rate of consumption with no change in the rate at which licking decayed across the test session. In contrast, intra-MR muscimol injections had little effect on the initial licking rate, but greatly increased meal duration, indicating that this treatment affected ingestion in a different way than did altering the sucrose concentration. Muscimol injections produced a significantly larger increase in the intake of the 0.29 M than of the 0.05 M solution. Intra-MR muscimol injections did not alter the within burst rate of licking, suggesting that they did not affect the functioning of the licking pattern generator. In contrast, these injections did increase the number of licks contained within “clusters,” that is groups of licks separated from each other by intervals of more than 0.5 sec. These findings show that inactivation of the MR produces a powerful effect on the intake of liquid diets, and that the nature of this effect is different from that produced here by changes in sucrose concentration and from those reported after pharmacological manipulations of a number of other brain systems. We additionally discuss several theoretical issues arising in the interpretation of microstructural data. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

Different functional effect of Ro 15-4513 and Ro 19-4603 on synaptic responses of Purkinje cells in the rat cerebellar slice

The Sugiura operation has been reported to have low operative mortality, rebleeding, and encephalopathy rates when carried out in a predominantly nonalcoholic Japanese population with good liver function. A literature review of reports of the Sugiura procedure outside Japan reveals a high complication and mortality rate when it is used as an emergency procedure in patients with advanced liver disease, especially in those with alcoholic cirrhosis. Uncontrolled studies report results that differ little from the Japanese series when the operation is confined to good-risk patients in the elective situation. Our experience with the Sugiura operation supports its role in these circumstances, especially in patients with portal vein thrombosis and normal liver function. The only good prospective controlled trial has been carried out in patients with schistosomiasis and suggests that the Sugiura operation is far superior to total shunt and may have a slight advantage over the Warren shunt because of its low incidence of postoperative encephalopathy. More controlled trials are required to establish its role in good- to moderate-risk patients with alcoholic cirrhosis.     

The effects of the novel benzodiazepine receptor inverse agonist Ru 34000 on ethanol-maintained behaviors

(1) In vagotomized, anaesthetized rats, effects of stimulation of cardiac N. vagus (2-25 Hz) on cardiac and circulatory functions were studied: we recorded transient reductions in heart rate (HR), in left-ventricular systolic pressure (LV Ps), in maximal change in left-ventricular pressure development (dp/dt)max and in mean arterial pressure (MAP, A. femoralis). (2) Bolus injection of angiotensin II (AII, 2.5-100 microg/kg body weight) caused (a) transient increases in HR, LV Ps and MAP (pressor effects, maximal changes occurred within 3 min after injection), and (b) dose-dependently reduced effects of vagus stimulation (non-pressor effects, recorded 10 min after injection). Due to fast breakdown of All in the circulatory system, all observed vagus stimulation effects were completely recovered within 1 h after injection. (3) Plasma concentration of AII was recorded with a highly specific radioimmunoassay: 10 min after AII injection (non-pressor range), plasma concentration was clearly higher than physiological levels in all experiments with 10 microg AII/kg at least. (4) Treatment with propranolol (beta-adrenoceptor blocker, 1 mg/kg body weight) did not reduce the vagus effects alone, but decreased the modulatory AII effects. This result hints at the activation of sympathetic beta-adrenergic receptors by AII counteracting the parasympathetic cardiac control.     

Effects of buprenorphine and Ro 15-4513 on delayed death and brain beta-endorphin levels in rats treated with cocaine or cocaine-ethanol

The present study was aimed at elucidating the relationship between brain beta-endorphin, which was estimated by the immunofluorescence method, and fatal drug toxicities due to cocaine and combined cocaine-ethanol administration, including the late fatal toxicities clinically noted. beta-endorphin is an endogenous opioid peptide, and its secretion has been suggested to be influenced by physiological stresses. Furthermore, since protection against these fatal toxicities has been previously reported to be provided by buprenorphine (a ligand for opioid receptors) and Ro 15-4513 (a ligand for benzodiazepine receptors), this study also focused on the relationship between the effects of these two ligands and the changes in brain beta-endorphin immunoreactivity. In the fatal toxicity study, a toxic dose (75 mg/kg, i.p.) of cocaine combined with and without ethanol (3 g/kg, i.p.) was administered to the rats, with and without buprenorphine (0.25, 0.5, 1 mg/kg, i.p.) or Ro 15-4513 (5, 10, 15 mg/kg, i.p.). All of the deaths that occurred in these animals were divided into two groups: early deaths with early toxic symptoms in which the drugs were detected in the tissue samples, and late deaths with late toxic symptoms in which no drugs were detected in the samples. Without the administration of buprenorphine or Ro 15-4513, the frequency of late deaths was higher in the cocaine group as compared to the cocaine-ethanol group. The total mortality rate was effectively attenuated by treatment with 0.25 mg/kg buprenorphine or 10 mg/kg Ro 15-4513. Following treatment with 1 mg/kg buprenorphine or 15 mg/kg Ro 15-4513, the frequency of late deaths was significantly enhanced in the cocaine group. The brain and liver cocaethylene concentrations were also attenuated in those groups in which the total mortality rates were attenuated. In the brain beta-endorphin immunoreactivity study, the number of beta-endorphin immunoreactive nerve cells at the arcuate nucleus was counted at 3 minutes or 24 hours after the drug treatment. At 3 minutes after the drug treatment, the number of weakly immunoreactive cells with photographic light absorption values greater than 50% was enhanced in the groups in which the frequency of late deaths had been increased. In the cocaine-ethanol groups treated with buprenorphine or Ro 15-4513, this enhancement of weakly immunoreactive cells was observed when the total mortality rate was increased, regardless of the type of death. At 24 hours after the drug treatment (50 mg/kg cocaine), an enhancement of the weakly immunoreactive cells only was observed in all of the groups in which the occurrence of toxicities had been enhanced, regardless of the type of toxicity. Therefore, it can be concluded that the enhancement of total brain beta-endorphin immunoreactivity was closely correlated with the increase in the frequency of total fatal toxicities, and that the enhancement of weakly immunoreactive cells was closely correlated with the increase in the frequency of delayed fatal toxicities.     

Tolerance to amphetamine hypophagia: A miscrostructural analysis of licking behavior in the rat.

The development of tolerance to amphetamine-induced hypophagia was assessed by recording changes in lick parameters in rats given chronic administration of the drug (2 mg/kg) and access to sweetened milk. Although licking and milk intake gradually recovered, the volume of milk ingested per lick remained suppressed. Amphetamine had no effect on the interlick interval or the force per lick. In contrast, the drug caused a sustained increase in the number of lick bursts (defined by pause criteria of 0.5-2.0 s) and a decrease in the number of licks per burst (but only at pause criteria of 0.5 and 1.0 s). These results suggest that tolerant rats frequently interrupt licking, resulting in less efficient capture of milk. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effects of the benzodiazepine inverse agonist RO19-4603 alone and in combination with the benzodiazepine receptor antagonists flumazenil, ZK 93426 and CGS 8216, on ethanol intake in alcohol-preferring (P) rats

The present study investigated dose dependence and time course effects of the benzodiazepine (BDZ) partial inverse agonist, RO19-4603 (0.005-0.30 mg/kg) alone, and in combination with the BDZ receptor antagonists flumazenil, ZK 93426, and CGS 8216 (20 mg/kg) in selectively bred alcohol-preferring (P) rats provided a two-bottle choice test between ethanol (EtOH) (10% v/v), and a palatable saccharin (0.0125% g/v) solution. A single dose of RO19-4603 as low as 0.009 mg/kg selectively reduced EtOH drinking during the initial 15 min of a 4 h access to 19-0% of control levels on day 1. The 0.08, 0.15 and 0.30 mg/kg doses of RO19-4603 significantly reduced total EtOH intake in the 4 h access period to 57-45% of controls on day 1. On day 2, no RO19-4603 injections were given; however, six of the seven doses of RO19-4603 (0.009, 0.02, 0.04, 0.08, 0.15, and 0.30 mg/kg) continued to reduce EtOH intake to 42-3% of control levels at the initial 15 min interval, while the 0.005, 0.009, 0.08, and 0.30 mg/kg doses reduced total 4 h EtOH intake to 60-42% of controls. Saccharin intake was either not altered by RO19-4603 or showed increases during the initial 15 min intervals and the total 4 h sessions on days 1 and 2. Food intake was also unaffected by RO19-4603. The CGS 8216, but neither flumazenil nor ZK 93426, reliably reversed the RO19-4603-induced suppression of EtOH intake on days 1 and 2. That certain BDZ inverse agonists can attenuate motivated behavior for EtOH reinforcement over a prolonged time course may provide a possible therapeutic approach to reducing EtOH consumption associated with alcoholism.     

The dorsal raphe nucleus is a site of action mediating the behavioral effects of benzodiazepine receptor inverse agonist DMCM.

Systemic administration of benzodiazepine receptor inverse agonists leads to behavioral changes similar to those produced by inescapable shock (IS). The dorsal raphe nucleus (DRN) is a critical structure mediating IS effects. The present experiments determined whether the DRN is a site mediating the behavioral changes produced by benzodiazepine receptor inverse agonists. Microinjection of the inverse agonist Methyl 6,7-Dimethoxy-4-ethyl-β-carboline-3-carboxylate (DMCM) in the region of the DRN produced enhancement of fear conditioning as assessed by the amount of freezing in the presence of shock cues as well as interference with shuttlebox escape learning assessed 24 hr later. Furthermore, lesion of the DRN blocked the effects of systemic DMCM on fear conditioning and escape learning. These data suggest that the DRN is indeed critical in mediating these behavioral consequences of DMCM and further support a role for the DRN in producing the behavioral changes induced by IS. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Punishment of consummatory and instrumental behavior: Effects on licking and bar pressing in rats.

Maintained licking and bar pressing in 48 male hooded rats by intermittent water reinforcement. Shock punishment was delivered for either a reinforced or nonreinforced lick or bar-press response. Punishment suppressed licking more than bar pressing. Pairing punishment with reinforcement had little effect on overall response suppression. In Exp. II with 18 Ss, Ss punished for the 1st lick after a reinforced bar press showed more response suppression than Ss punished for the reinforced bar press. Results support R. Solomon's (see record 1965-00694-001) hypothesis than consummatory behavior is more sensitive to punishment than instrumental behavior. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Changes in the rat's preference for saccharin and sodium chloride solutions following injection of alloxan monohydrate.

Conducted 5 experiments with a total of 136 male Sprague-Dawley rats in which alloxan-diabetic rats were compared with saline-injected controls on 2-choice preference tests involving saccharin and water or NaCl and water. Previous results indicating a decrease in preference for saccharin were confirmed and extended. However, a similar pattern of preference change for NaCl was not found. Results indicate that saccharin and NaCl consumption were not affected in the same way by experimental diabetes and that the loss of palatability of saccharin during a chronic diabetic state was not consistent with learning models of taste-aversion formation. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Analytical issues in the evaluation of food deprivation and sucrose concentration effects on the microstructure of licking behavior in the rat

A microstructural analysis of licking behavior in nondeprived and 23-hr food-deprived rats (n = 15) presented with various sucrose solutions (0.03-1.0 M) in daily single-bottle, 1-hr sessions was conducted. Food deprivation and concentration interacted to increase total licks. The effects of food deprivation and concentration on burst size (BS), burst number (BN), and other parameters varied as a function of the pause criterion (PC; 0.3-100 s) used to define licking bursts. A rationale for selecting a 1-s PC for further analysis is presented. Despite the lack of correlations between temporally contiguous burst and pause combinations, mean BS decreased and pause duration increased as meals progressed. At the 1-s PC, BS increased linearly with concentration, implying that this microstructural parameter is influenced in part by taste. Food deprivation did not affect BS but rather increased BN and proportionally extended the meal duration.     

The benzodiazepine inverse agonist DMCM as an unconditional stimulus for fear-induced analgesia: Implications for the role of GABAA receptors in fear-related behavior.

When the benzodiazepine inverse agonist DMCM (6,7-dimethoxy-4-ethyl-β-carboline-3-carboxylic acid methyl ester) occupies the benzodiazepine recognition site on the GABAA receptor complex, the inhibitory action of γ-aminobutyric acid (GABA) is attenuated. DMCM acted as an unconditioned stimulus (UCS) for 1 response associated with fear or anxiety, analgesia, as indicated by a dose-dependent (0.25–2.0 mg/kg) suppression of rats' responses to a formalin injection. This was accompanied by other fearlike responses (defecation and urination). The opioid antagonist naltrexone (1.75–24 mg/kg) did not affect these behaviors. Environmental cues associated with DMCM provoked analgesia and defecation in the absence of the drug. The conditional analgesia was reversed by naltrexone (7 mg/kg). DMCM functions as an unconditional fear stimulus by eliciting fear-related behaviors and conditioning those responses to neutral stimuli. The neural circuitry underlying fear conditioning appears to involve tonically inhibitory GABAergic synapses. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Imaging of the super high affinity binding sites for [3H]Ro15-4513 in rat hippocampus: comparison between in vitro and in vivo binding

Using fluorescence optical and electron spin resonance spectroscopy, we have investigated the production of superoxide by bovine endothelial nitric oxide synthase (NOS). In contrast to neuronal NOS, the heme moiety is identified as the exclusive source of superoxide production by endothelial NOS. Thus, calmodulin-mediated enzyme regulation affects production of nitric oxide and superoxide simultaneously and inseparably. The balance between the nitric oxide/superoxide reaction pathways may be shifted by addition of exogenous heme-specific agents, such as tetrahydrobiopterin. Our results have direct relevance for the pathophysiology of atherosclerosis.     

The consequences of gustatory nerve transection on taste-guided licking of sucrose and maltose in the rat.

Lick responses to sucrose and maltose (0.01–1.0 M) were measured in nondeprived rats during brief-access taste trials before and after histologically confirmed gustatory neurotomy. Pronounced decreases in sugar responsiveness occurred after combined section of the chorda tympani (CT) and greater superficial petrosal nerves. The additional section of the glossopharyngeal nerve (GL) flattened the sucrose concentration-response function. Extirpation of the sublingual and submaxillary salivary glands also attenuated sugar responsiveness. Section of the CT or GL alone or in combination caused less severe or no decreases in sugar licking. There were signs of licking impairments after some of these neurotomies, but the data suggest that changes in sugar responsiveness were not solely motor in origin. Thus the 7th nerve is necessary and most likely sufficient for the maintenance of normal unconditioned appetitive responsiveness to sucrose and maltose. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effects of sugar ingestion on the classroom and playgroup behavior of attention deficit disordered boys.

16 attention deficit disordered (ADD) boys (aged 6.4–9.1 yrs) fasted overnight and then received in the morning a challenge drink containing either 1.75 g/kg sucrose or a placebo (aspartame) of comparable sweetness. Ss received sugar and placebo 2 days each. Dependent variables consisted of measures of classroom behavior, academic productivity and accuracy, noncompliance with adult requests, and positive and negative peer interactions. Results offer no support for the contention that sugar ingestion adversely affects the behavior or learning of ADD boys. Limitations of the present study, including the dosage of sugar employed and the use of concurrent interventions, are discussed. It is suggested that beliefs that sugar has a challenge effect on behavior in ADD children may actually reflect their difficulty readjusting to classroom activity after a snack period. (32 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Self-administration of ethanol and saccharin in newborn rats: Effects on suckling plasticity.

Responsiveness to a surrogate nipple providing water, saccharin, 5% ethanol, or 10% ethanol was tested in newborn rats naive to suckling (3–5 hr old) on Postnatal Day (P) 0 and in older neonates with regular suckling experience on P1 or P2. At all ages, pups demonstrated greater nipple attachment for saccharin or 5% ethanol than for water. Intake of saccharin and 5% ethanol was high in newborns, far exceeding that of water. At P1 and P2, pups exhibited a preference for saccharin, but not for 5% ethanol. Preexposure to a nipple providing ethanol or saccharin (but not a nipple alone or fluids alone) increased subsequent responsiveness toward an empty surrogate nipple (no fluid), both 1 hr and 24 hr after preexposure. Although, during preexposure, pups responded most positively to the nipple providing saccharin, the longest time spent on an empty nipple was observed in pups preexposed to 10% ethanol. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Molecular structure and stereoelectronic properties of sarmazenil--a weak inverse agonist at the omega modulatory sites (benzodiazepine receptors): comparison with bretazenil and flumazenil

X-ray diffraction and ab initio MO theoretical calculations were used in order to investigate the structural and electronic properties of sarmazenil, a weak inverse agonist at the omega modulatory sites (benzodiazepine receptors). This compound was compared to bretazenil, a partial agonist, and to the antagonist flumazenil on the basis of structural and electronic data. The conformational and theoretical properties (interatomic pi overlap populations, molecular electrostatic potential (MEP), the topology of frontier orbitals, and proton affinity) of these three imidazobenzodiazepinones were determined in order to analyse the stereoelectronic properties in relation with their distinct intrinsic efficacies at the omega modulatory sites.     

Additivity of taste-specific effects of sucrose and quinine: Microstructural analysis of ingestive behavior in rats.

Effects of sweet and bitter tastes on ingestion were studied by timing licking responses. 12 water-deprived rats were given 15-min access to sucrose (SU) solutions (0.00, 1.25, 2.50, and 5.00%) with and without quinine (Q; 0.01%) and to Q solutions (0.00, 0.0025, 0.005, and 0.01%) with and without SU (5.00%). Volume ingested and number of licks increased with SU and decreased with Q. In response to SU, the number of bursts increased, and interlick intervals lengthened. In response to Q, licks to ingest 1 ml of solution, burst number, and percentage of slow licks increased, and burst size decreased. When Q and SU were mixed in the same solution, the pattern of ingestive responses manifested attributes of both tastes. Results suggest 2 separate, parallel systems that operate simultaneously to govern rats' licking behavior. One system expresses the effects of SU on the pattern of ingestion and the other expresses the effects of Q. (PsycINFO Database Record (c) 2010 APA, all rights reserved)